Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

OBJECTIVE To explore the effect and safety of dapagliflozin on cardiac function and renal function， blood glucose， and quality of life in patients with heart failure （HF） combined with chronic kidney disease （CKD）. METHODS A total of 156 patients with HF combined with CKD admitted to Shangqiu First People’s Hospital from January 1， 2021 to January 1， 2023 were included. According to the random number table， the patients were randomly divided into conventional treatment group （n＝80） and dapagliflozin group （n＝76）. Conventional treatment group was given conventional treatment； dapagliflozin group was additionally given Dapagliflozin tablets 10 mg orally， once a day， based on conventional treatment group. Both groups were treated for 6 months. Cardiac function [left ventricular ejection fraction （LVEF）， left ventricular end-systolic diameter （LVESD）， left ventricular end-diastolic diameter （LVEDD）， N-terminal pro-brain natriuretic peptide （NT-proBNP）]， renal function [blood creatinine， urea nitrogen， urinary albumin excretion rate （UAER）， glomerular filtration rate （GFR）， creatinine 806731979@qq.com clearance rate （CCR）]， glycosylated hemoglobin， and the quality of life were observed in 2 groups before and after treatment. The occurrence of ADR was recorded. RESULTS After treatment， LVESD， LVEDD， NT-proBNP， blood creatinine， urea nitrogen， UAER in 2 groups as well as the level of glycosylated hemoglobin in dapagliflozin group were significantly lower than before treatment； the dapagliflozin group was significantly lower than the conventional treatment group. LVEF， GFR， CCR and Kansas City Cardiomyopathy Questionnaire score were significantly higher than before treatment， and the dapagliflozin group was significantly higher than the conventional treatment group （P＜0.05）. There was no statistical significance in glycosylated hemoglobin of conventional treatment group before and after treatment （P＞ 0.05）. There was no statistically significant difference in the incidence of dizziness， rash， liver dysfunction， urinary system infection， new dialysis and hypotension between the two groups （P＞0.05）. CONCLUSIONS Dapagliflozin can improve the cardiac function and renal function of patients with HF complicated with CKD， improve patients’ quality of life and lower blood sugar levels without increasing the risk of adverse events.

Objective To understand the changing distribution and antimicrobial resistance profiles of Proteus,Morganella and Providencia in hospitals across China from January 1,2015 to December 31,2021 in the CHINET Antimicrobial Resistance Surveillance Program.Methods Antimicrobial susceptibility testing was carried out following the unified CHINET protocol.The results were interpreted in accordance with the breakpoints in the 2021 Clinical & Laboratory Standards Institute(CLSI)M100(31 st Edition).Results A total of 32 433 Enterobacterales strains were isolated during the 7-year period,including 24 160 strains of Proteus,6 704 strains of Morganella,and 1 569 strains of Providencia.The overall number of these Enterobacterales isolates increased significantly over the 7-year period.The top 3 specimen source of these strains were urine,lower respiratory tract specimens,and wound secretions.Proteus,Morganella,and Providencia isolates showed lower resistance rates to amikacin,meropenem,cefoxitin,cefepime,cefoperazone-sulbactam,and piperacillin-tazobactam.For most of the antibiotics tested,less than 10％of the Proteus and Morganella strains were resistant,while less than 20％of the Providencia strains were resistant.The prevalence of carbapenem-resistant Enterobacterales(CRE)was 1.4％in Proteus isolates,1.9％in Morganella isolates,and 15.6％in Providencia isolates.Conclusions The overall number of clinical isolates of Proteus,Morganella and Providencia increased significantly in the 7-year period from 2015 to 2021.The prevalence of CRE strains also increased.More attention should be paid to antimicrobial resistance surveillance and rational antibiotic use so as to prevent the emergence and increase of antimicrobial resistance.

Objective: To explore the neuroprotective effects of the Shaoyao Gancao decoction (SGD) against excitatory damage in PC12 cells and the role of the Src-NR2-nNOS pathway mediation by SGD in regulating γ-aminobutyric acid (GABA)-glutamate (Glu) homeostasis. Methods: N-Methyl-d-aspartic acid (NMDA) was used to establish a PC12 cell excitability injury model. To investigate the neuroprotective effect of SGD, a cell counting kit-8 (CCK-8) assay was used to determine PC12 cell viability, Annexin V/Propidium Iodide (Annexin V/PI) double staining was used to determine PC12 cell apoptosis, and Ca2+ concentration was observed using laser confocal microscopy. GABA receptor agonists and antagonists were used to analyze the neuroprotective interactions between γ-aminobutyric acid (GABA) and NMDA receptors. Additionally, molecular biology techniques were used to determine mRNA and protein expression in the Src-NR2-nNOS pathway. We analyzed the correlations between the regulatory sites of GABA and NMDA interactions, excitatory neurotoxicity, and brain damage at the molecular level. Results: NMDA excitotoxic injury manifested as a significant decrease in cell activity, increased apoptosis and caspase-3 protein expression, and a significant increase in intracellular Ca2+ concentration. Administration of SGD, a GABAA receptor agonist (muscimol), or a GABAB receptor agonist (baclofen) decreased intracellular Ca2+ concentrations, attenuated apoptosis, and reversed NMDA-induced upregulation of caspase-3, Src, NMDAR2A, NMDAR2B, and nNOS. Unexpectedly, a GABAA receptor antagonist (bicuculline) and a GABAB receptor antagonist (saclofen) failed to significantly increase excitatory neurotoxicity. Conclusions Taken together, these results not only provide an experimental basis for SGD administration in the clinical treatment of central nervous system injury diseases, but also suggest that the Src-NR2A-nNOS pathway may be a valuable target in excitotoxicity treatment.

Objective:To investigate the effects of using a high monounsaturated fatty acid (MUFA) and low carbohydrate formula on blood glucose levels and diarrhea treatment effects in critically ill neurological patients.Methods:A self-controlled before-and-after study design was employed, with 13 patients admitted to the neurology intensive care unit (ICU) of the First Affiliated Hospital of Sun Yat-sen University from November to December 2023, who were treated with a high MUFA and low carbohydrate formula [Glucerna enteral nutrition (EN) preparation]. Changes in blood glucose parameters within 7 days before and after the use of Glucerna EN preparation were analyzed, including standard deviation ( SD) of blood glucose, mean blood glucose (MG), median blood glucose, mean amplitude of glycemic excursions (MAGE), largest amplitude of glycemic excursions (LAGE), coefficient of variation ( CV) of blood glucose, the incidence of hyperglycemia (> 7.8 mmol/L) and severe hyperglycemia (> 13.9 mmol/L), and daily insulin dose. Changes in total protein (TP), albumin (ALB), hemoglobin (Hb), C-reactive protein (CRP), and white blood cell count (WBC) were observed before and after intervention. Improvement in diarrhea symptoms, Hart diarrhea score, Bristol Stool classification score, and incontinence dermatitis classification were also analyzed before and after the use of Glucerna EN preparation. Results:A total of 13 critically ill neurological patients were enrolled, among whom 9 patients had a history of hyperglycemia and 8 patients had diarrhea symptoms. After intervention with Glucerna, the patients' SD of blood glucose, MG, median blood glucose, MAGE, LAGE, CV of blood glucose, incidence of hyperglycemia, incidence of severe hyperglycemia, and daily insulin dose were all lower than those before the intervention [ SD of blood glucose (mmol/L): 1.83±1.11 vs. 2.10±1.13, MG (mmol/L): 8.87±2.03 vs. 9.75±1.37, median blood glucose (mmol/L): 9.12±1.67 vs. 10.17±0.48, MAGE (mmol/L): 0.66±0.31 vs. 0.78±0.32, LAGE (mmol/L): 4.95±3.64 vs. 5.58±3.10, CV of blood glucose: 16.00% (11.00%, 28.50%) vs. 18.00% (12.50%, 27.50%), hyperglycemia incidence: 47.31% vs. 74.66%, severe hyperglycemia incidence: 6.08% vs. 6.71%, daily insulin dose (U): 5.25 (0.00, 32.59) vs. 20.76 (0.00, 66.88)], with a significant decrease in daily insulin dose after the intervention ( P < 0.05); TP, ALB, Hb, CRP and WBC showed no significant changes before and after the intervention with Glucerna EN preparation. The improvement time of diarrhea symptoms after intervention was (3.50±1.41) days, and the Hart diarrhea score on the seventh day after intervention (4.88±3.48 vs. 10.00±3.38) and the Bristol Stool classification score on the third and seventh days after intervention (5.87±0.35, 5.50±0.53 vs. 6.50±0.53) were significantly lower than before the intervention (all P < 0.05). Before the intervention with Glucerna EN preparation, the classification of incontinence dermatitis was mainly classified as Grade 2 severity (71.43%); after the intervention, it significantly improved by the seventh day, with Grade 1 being the main classification (57.14%). Conclusion:The high MUFA and low carbohydrate formula has a positive effect on blood glucose control and diarrhea treatment in critically ill neurological patients.

Objective To investigate whether activation of mitochondrial acetal dehydrogenase 2(ALDH2)alleviates hypoxic pulmonary hypertension by regulating the SIRT1/PGC-1α signaling pathway.Methods Thirty 8-week-old C57 BL/6 mice were randomized into control,hypoxia,and hypoxia+Alda-1(an ALDH2 activator)group(n=10),and the mice in the latter two groups,along with 10 ALDH2 knockout(ALDH2-/-)mice,were exposed to hypoxia(10%O2,90%N2)with or without daily intraperitoneal injection of Alda-1 for 4 weeks.The changes in right ventricular function and pressure(RVSP)of the mice were evaluated by echocardiography and right ventricular catheter test,and pulmonary artery pressure was estimated based on RVSP.Pulmonary vascular remodeling,right ventricular injury,myocardial α-SMA expression,distal pulmonary arteriole muscle normalization,right ventricular cross-sectional area,myocardial cell hypertrophy,and right cardiac hypertrophy index were assessed with HE staining,immunofluorescence staining and WGA staining,and the expressions of ALDH2,SIRT1,PGC-1α,P16INK4A and P21CIP1 were detected.In pulmonary artery smooth muscle cells with hypoxic exposure,the effect of Alda-1 and EX527 on cell senescence and protein expressions was evaluated using β-galactose staining and Western blotting.Results The wild-type mice with hypoxic exposure showed significantly increased RVSP,right ventricular free wall thickness and myocardial expressions of P16INK4A and P21CIP1,which were effectively lowered by treatment with Alda-1 but further increased in ALDH2-/-mice.In cultured pulmonary artery smooth muscle cells,hypoxic exposure significantly increased senescent cell percentage and cellular expressions of P16INK4A and P21CIP1,which were all lowered by treatment with Alda-1,but its effect was obviously attenuated by EX527 treatment.Conclusion ALDH2 alleviates hypoxia-induced senescence of pulmonary artery smooth muscle cells by upregulating the SIRT1/PGC-1α signaling pathway to alleviate pulmonary hypertension in mice.

Objective:To clarify the effects of microwave radiation on anxiety-like behavior, the histomorphology of the secondary visual cortex, and calcium activity in neurons.Methods:36 C57BL/6N mice were selected and divided into control group and microwave radiation group according to the random number table method. In the simple behavioral testing, there were 8 mice in the control group and 7 mice in the radiation group. Combining fiber optic recording with behavioral experiments, there were 8 mice in the control group and 7 mice in the radiation group. Hematoxylin-eosin (HE) staining was conducted with 3 mice in each group. A high-power microwave simulated source in the X-band with a center frequency of 9.875 GHz and an average power density of 12 mW/cm 2 was used to irradiate the mice for 15 minutes, establishing a microwave radiation animal model. Then, anxiety-like behavior changes in the radiation group were identified using the open-field and elevated plus maze (EPM) tests. The effects of microwave radiation on the histomorphology of the secondary visual cortex were investigated using HE staining and optical microscopy. Based on the genetically encoded calcium imaging technique, as well as optical fiber recording combined with behavioral paradigms in the open field and the EPM, the changes of calcium activity in neurons in the V2M region of the secondary visual cortex were detected. Results:Compared to the control group, the radiation group showed a significant decrease in the frequency of exploring the central region of the open field and the open arm of the EPM ( t = 2.24, 3.10, P < 0.05). Furthermore, the radiation group exhibited the degeneration and apoptosis of some neurons in the secondary visual cortex, primarily manifested as pyknosis and deep staining, cell body shrinkage, and the slightly widening of perivascular space. Fiber optic recordings and behavioral experiments indicated that compared to the control group, mice in the radiation group exhibited significantly increased calcium activities in neurons of the secondary visual cortex when exploring the central region of the open field ( t = -2.75, P < 0.05) or the open arm of the EPM ( t = -2.77, -3.41, P < 0.05) compared to those before radiation after microwave exposure. Conclusions:Microwave radiation can induce anxiety-like behaviors and histopathological changes in the secondary visual cortex. Increased calcium activity in neurons of the secondary visual cortex is proved to be an important mechanism underlying the changes in anxiety-like behavior due to microwave radiation.

Objective:To investigate the effect of different concentrations and infection durations of varicella-zoster virus (VZV) on nucleotide-binding and oligomerization domain-like receptor protein 3 (NLRP3) inflammasome-related proteins in mouse brain astrocytes.Methods:Mouse brain astrocytes were infected with different dilutions (10 0 [undiluted original concentration] - 10 -7) of cell-free VZV suspension, and the cytopathic effect was observed under a microscope in each group to determine the concentration and infection duration of VZV for subsequent experiments. Some mouse brain astrocytes were infected with cell-free VZV suspension at dilutions of 10 0, 10 -2, 10 -4, and 10 -6, and samples were collected on day 5; some other mouse brain astrocytes were infected with cell-free VZV suspension at the median tissue culture infective dose (TCID 50) of 1.0 × 10 5/ml, and the samples were collected on days 3, 5, 7, and 9, separately; normally cultured mouse brain astrocytes served as control groups in the above experiments. Western blot analysis was performed to determine the protein expression of VZV glycoprotein E (gE), NLRP3, caspase-1, and interleukin-1β (IL-1β) in mouse brain astrocytes in each group. One-way analysis of variance was used for comparisons of data with a normal distribution, and least significant difference- t test was used for multiple comparisons. Results:On day 3 after infection with VZV at the dilution of 10 0, the number of cells decreased, some cells became irregular, lost their original shapes, and showed granular cytoplasm and an unclear nucleus structure, while some cells fused and formed multinucleated giant cells; on day 5, ≥ 50% of cells showed the aforementioned changes and some intercellular spaces were dilated giving a network-like appearance in the 10 0-dilution group; on day 9, the 10 0- - 10 -6- dilution groups all showed dilated intercellular spaces arranged in networks and cytopathic effect in ≥ 50% of cells; on day 10, some cells in the control group exhibited cytoplasmic vacuolation, exfoliation and flotation. Based on these results, the following experiments were conducted with VZV at a concentration gradient ranging from 10 0 to 10 -6 dilutions, and the observations were performed from day 3 to day 9. Western blot analysis showed increased protein expression of VZV gE, NLRP3, caspase-1, and IL-1β in the VZV infection groups compared with the control group, and there were significant differences among different VZV concentration groups ( F = 21.45, 43.82, 21.85, 29.53, respectively, all P < 0.05) and different infection duration groups ( F = 224.10, 78.79, 186.50, 85.08, respectively, all P < 0.05). With the increasing dilution ratios, the expression of inflammasome-related proteins (NLRP3, caspase-1, and IL-1β) decreased, with the highest expression observed in the 10 0-dilution group and the lowest in the 10 -6-dilution group (all P < 0.05). On days 3, 5, and 7 after VZV infection, the expression of NLRP3 inflammasome-related proteins increased over time, with the highest expression observed on day 7 and the lowest on day 3 (all P < 0.05), while their expression decreased on day 9. Conclusion:The expression of NLRP3 inflammasome-related proteins was up-regulated in mouse brain astrocytes after VZV infection, and their expression levels increased along with the increase in the VZV concentration and infection duration in a certain range.

Objective:To explore the characteristics of anorectal motility and sensation in patients with functional anorectal pain (FAP) by high-resolution anorectal manometry (HR-ARM) .Methods:The clinical data of 81 FAP patients (FAP group) who underwent HR-ARM in Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine from January 1, 2020 to January 31, 2022 were retrospectively collected, and 80 healthy volunteers were recruited as healthy control group during the same period. The HR-ARM characteristics were compared between FAP group and the healthy control group, between the patients with different genders in the FAP group, the patients with different subtypes (proctalgia fugax, levator syndrome, and non-specific FAP) in the FAP group, which included anal resting pressure, anal squeeze pressure, rectal pressure during simulated defecation, anal residual pressure during simulated defecation, paradoxical contractions, initial sensation threshold, defecation threshold, defecation urgency threshold, and tolerance threshold. Visual analogue scale (VAS) was used to assess the pain level of the patients in the FAP group, and Spearman correlation analysis was used to analyze the correlation between VAS and HR-ARM characteristics. Independent sample t-test, least significant difference test, Tamhane′s T2 test, and Mann-Whitney U test were used for statistical analysis. Results:The anal resting pressure, anal squeeze pressure, anal residual pressure during simulated defecation, defecation urgency threshold, and tolerance threshold of the FAP group were all lower than those of the healthy control group ((59.56±24.71) mmHg (1 mmHg=0.133 kPa) vs. (81.94±15.87) mmHg, (119.04±46.94) mmHg vs.(154.62±37.95) mmHg, 59.00(40.75, 80.95) mmHg vs. 83.10(61.78, 94.30) mmHg, 70.00(55.00, 90.00) mL vs. 85.00(60.00, 110.00) mL, 105.00(87.50, 150.00) mL vs. 140.00(100.00, 180.00) mL), and the differences were all statistically significant ( t=-6.83 and -5.29, Z=-4.12, -3.12 and -2.82; all P<0.01).The rectal pressure during simulated defecation of male patients in the FAP group was higher than that of males in the healthy control group, and the defecation urgency threshold was lower than that of males in the healthy control group (42.40(29.60, 57.95) mmHg vs. 31.10(25.85, 36.80) mmHg, 80.00(62.50, 107.50) mL vs. 92.00(81.00, 140.00) mL), and the differences were statistically significant ( Z=-1.99 and -2.53, both P<0.05). The anal resting pressure, anal squeeze pressure, anal residual pressure during simulated defecation, defecation urgency threshold, and tolerance threshold of female patients in FAP group were all lower than those of female in the healthy control group ((55.67±21.61) mmHg vs. (87.04±15.54) mmHg, (102.70±37.09) mmHg vs. (155.98±31.44) mmHg, 52.55(40.53, 67.48) mmHg vs. 83.10(61.10, 94.50) mmHg, 60.00(52.50, 81.50) mL vs. 80.00(60.00, 100.00) mL, 101.00(80.00, 128.75) mL vs. 120.00(94.00, 155.00) mL), and the differences were statistically significant ( t=-8.77 and -8.16, Z=-4.57, -2.24 and -2.14; all P<0.05). The anal resting pressure, anal squeeze pressure, anal residual pressure during simulated defecation, incidence rate of paradoxical contractions, defecation urgency threshold, and tolerance threshold of female patients in FAP group were all lower than those of male patients in FAP group ((55.67±21.61) mmHg vs. (68.28±29.16) mmHg, (102.70±37.09) mmHg vs. (155.62±46.66) mmHg, 52.55(40.53, 67.48) mmHg vs. 79.00(59.55, 99.25) mmHg, 28.6%(16/56) vs. 68.0%(17/25), 44.00(35.00, 60.00) mL vs. 60.00(45.00, 70.00) mL, 60.00(52.50, 81.50) mL vs. 80.00(62.50, 107.50) mL), and the differences were statistically significant( t=2.17 and 5.47, Z=-2.96, χ2=11.10, Z=-2.93 and -2.34; all P<0.05). The anal squeeze pressure of patients with proctalgia fugax subtype was higher than that of patients with levator syndrome subtype ((140.19±56.51) mmHg vs. (80.56±30.79) mmHg), and the tolerance threshold was lower than that of patients with non-specific FAP subtype ((87.86±17.80) mL vs. (125.14±48.31) mL), and the differences were statistically significant ( t=2.35 and 2.02, both P<0.05). The results of Spearman correlation analysis showed that VAS was negatively correlated with anal resting pressure, anal squeeze pressure, and defecation urgency threshold in the patients of the FAP group ( r= -0.28, -0.23, and -0.24; all P< 0.05). Conclusion:The presence of anorectal dismotility and sensory dysfunction in FAP may be related to pelvic floor muscle abnormalities, muscle coordination disorders during defecation, and rectal hypersensitivity.

Objective To investigate the effect of T cell immunoreceptor with Ig and ITIM domains (TIGIT) on the function of CD8⁺ T cells in the lungs of Plasmodium infected mice. Methods The lungs of the mice infected with Plasmodium yoelii were isolated, weighed and photographed after 12 days' infection. After dissolution, lung lymphocytes were isolated, counted and stained, and then the contents of CD8⁺ and TIGIT⁺CD8⁺ T cells were detected by flow cytometry. The expressions of L selectin (CD62L), CD69, programmed death 1 (PD-1), CD25, and C-X3-C motif chemokine receptor 1 (CX3CR1) on TIGIT⁺CD8⁺ T cells were detected by flow cytometry. After stimulation with phorbol 12-myristate 13-acetate (PMA) and ionomycin, the ability of TIGIT⁺CD8⁺T cells to secrete interferon γ(IFN-γ), interleukin 21 (IL-21), IL-4, IL-17, and IL-10 was detected. Results The body mass of mice with Plasmodium infection was reduced. The lungs became darker, and the ratio of the lung mass to body mass was significantly increased. Compared with the normal mice, the percentages and absolute quantity of CD8⁺ and TIGIT⁺CD8⁺ T cells in the lungs of the infected mice were significantly increased. The percentage of TIGIT⁺CD8⁺ T cells expressing CD62L in the infected group was significantly lower, while the percentage of the CD69, PD-1, and CX3CR1 cells were significantly higher than that of TIGIT⁺CD8⁺ T cells from the normal mice. The percentages of TIGIT⁺CD8⁺ T cells secreting IL-21, IL-4, IL-17 and IL-10 cells in the infected group were significantly lower. Conclusion The lung lesions from mice with Plasmodium infection are obvious, the numbers of TIGIT⁺CD8⁺ T cells increase, and these cells express a variety of activation-related molecules, but the ability to secrete cytokines is reduced.
Animals ; Mice ; CD8-Positive T-Lymphocytes ; Cytokines/metabolism* ; Interferon-gamma/metabolism* ; Interleukin-10/metabolism* ; Interleukin-17/metabolism* ; Interleukin-4/metabolism* ; Lung/metabolism* ; Malaria/metabolism* ; Plasmodium yoelii/metabolism* ; Programmed Cell Death 1 Receptor/metabolism*