Chronic heart failure （CHF） is a group of complex clinical syndromes caused by abnormal changes in the structure and/or function of the heart due to various reasons， resulting in disorders of ventricular contraction and/or diastole. CHF is a condition where primary diseases such as coronary heart disease， hypertension and pulmonary heart disease recur frequently and persist for a long time， presenting blood stasis in meridians and collaterals， stagnation of water and dampness， and accumulation of Qi in collaterals. Its pathogenesis is complex and may involve myocardial energy metabolism disorders， oxidative stress responses， myocardial cell apoptosis， autophagy， inflammatory responses， etc. According to the theory of restraining hyperactivity to acquire harmony， we believe that under normal circumstances， the adenosine monophosphate-activated protein kinase （AMPK） signaling pathway functions normally， maintaining human physiological activities and energy metabolism. Under pathological conditions， the AMPK signaling pathway is abnormal， causing energy metabolism disorders， inflammatory responses， and myocardial fibrosis. Traditional Chinese medicine （TCM） can regulate the AMPK signaling pathway through multiple mechanisms， targets， and effects， effectively curbing the occurrence and development of CHF. It has gradually become a research hotspot in the prevention and treatment of this disease. Guided by the theory of TCM， our research group， through literature review， summarized the relationship between the AMPK pathway and CHF and reviewed the research progress in the prevention and control of CHF with TCM active ingredients， TCM compound prescriptions， and Chinese patent medicines via regulating the AMPK pathway. The review aims to clarify the mechanism and targets of TCM in the treatment of CHF by regulating the AMPK pathway and guide the clinical treatment and drug development for CHF.

Background Aluminum (Al) is a lightweight metal that is widely present in the environment and the human body. It has been documented to cause various adverse health effects including the suppression of humoral immunity. Objective To investigate the role of oxidative stress in Al-induced humoral immunity suppression and to evaluate the possible protective effects of iron supplementation on this process. Methods Adult C57BL/6J mice were exposed to Al at concentrations of 0, 200, or 800 μg·mL⁻¹ via drinking water for three consecutive months. The expression of major histocompatibility complex class Ⅱ (I-A), proliferating cell markr-67 (Ki-67), and 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) in splenic B cells was evaluated through flow cytometry. Splenic B cells from the mice treated with 800 μg·mL⁻¹ Al or the control were sorted and treated in vitro with glutathione (GSH), N-Acetyl-L-cysteine (NAC), or a control vehicle. After 24 h, the expression of I-A was evaluated; and the hydroxyl radical (·OH)-generating potential, ·OH production, malondialdehyde (MDA) production, and iron content were assessed using commercial kits. Sixteen mice treated with 800 μg·mL⁻¹ Al received an intravenous injection of either a ferric chloride solution containing 0.3 g·L⁻¹ iron or a 0.9% sodium chloride solution, while eight control mice received 0.9% sodium chloride solution; the injection volume was 0.1 mL per mouse. Two and a half days after injection, I-A and Ki-67 expressions, ·OH-generating potential, ·OH production, and MDA production in splenic B cells were measured; and the concentrations of serum immunoglobulin (Ig) M and IgG were measured through (enzyme-linked immunosorbent assay) ELISA. The splenic B cells sorted from untreated mice were exposed to 0, 12.5, 25, or 50 μg·L⁻¹ Al in vitro. The splenic B cells treated with 50 μg·L⁻¹ Al and the splenic B cells sorted from 800 μg·mL⁻¹ Al-treated mice were additionally treated with GSH and NAC in vitro. The iron supplementation groups, which included the 50 μg·L⁻¹ Al-treated group and splenic B cells sorted from 800 μg·mL⁻¹ Al-treated mice, were treated with a culture medium containing 30 μmol·L⁻¹ iron in vitro. I-A and Ki-67 expressions, ·OH-generating potential, ·OH production, and MDA production in B cells were detected after a 24-h treatment period. Results In the in vivo mouse model, exposure to 800 μg·mL⁻¹ Al significantly inhibited the I-A and Ki-67 expressions (P<0.05), increased DCFH-DA expression and ·OH-generating potential (P<0.05, P<0.01), decreased iron content (P<0.01) and ·OH and MDA production (P<0.01, P<0.001) of splenic B cells, as well as serum IgM and IgG concentrations (P<0.05, P<0.01) in the mice. Exposure to 200 μg·mL⁻¹ Al showed a tendency to decrease the I-A and Ki-67 expressions, and to increase the DCFH-DA expression in splenic B cells, but these differences were not significant. In the in vitro splenic B-cell model, Al (12.5, 25, and 50 μg·L⁻¹) inhibited I-A and Ki-67 expressions (P<0.05, P<0.01) across all concentrations; 50 μg·L⁻¹ Al increased the ·OH-generating potential (P<0.05), and decreased ·OH and MDA production (P<0.01, P<0.05) in B cells. Treatment with GSH and NAC further suppressed I-A expression (P<0.05) in B cells. Iron supplementation increased the ·OH and MDA production (P<0.05), restored I-A and Ki-67 expressions (P<0.05, P<0.01) in B cells, and elevated the serum IgM and IgG concentrations (P<0.05) in Al-treated mice. Conclusion Al suppresses humoral immunity and ·OH production in B cells. The underlying mechanism may involve the decreased iron content and the subsequent retardation of the Fenton reaction in B cells. Supplementing with iron can restore the Fenton reaction in B cells and potentially reverse Al-induced impairment of humoral immunity.

Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

Objective:To investigate the occurrence and managements of poor recovery after total endoscopic middle ear surgery. Methods:A total of 302 cases（315 ears） who underwent endoscopic middle ear surgery in our hospital from June 2020 to June 2021 were collected. Follow up by means of endoscopy, pure tone hearing threshold, tympanogram was conducted at 1 month, 3 months, 6 months and 1 year after surgery to analyze the incidence, possible causes, treatment strategies and effects of poor results tympanic membrane healing and hearing recovery. Results:Among 302 patients（315 ears） followed up, there were 28 cases with poor recovery. There were fourteen cases of poor eardrum healing, of which 10 cases achieved healing of eardrum after tympanic membrane patching in the outpatient department, with a success rate of about 71.4%. TM recurrence adhesion occurred in 4 cases after surgeries of cholesteatoma and adhesive otitis media. One case completely recovered after self eustachian tube insufflation, while 2 cases maintained the degree of eardrum subsidence, and one ineffective patient chose resurgical treatment, with an effective rate was 75.0%. Failure in hearing improvement occurred in 8 cases, all of which underwent second surgical exploration, and seven cases were improved after the second surgery, with an effective rate of 87.5%. Among the 8 patients with no improvement or aggravation of hearing loss after surgery, four cases had postoperative B-type or C-type of tympanogram, and the hearing could not improve after self eustachian tube insufflation for secondary surgical exploration. and the hearing improved after the secondary surgery. Incorrect orientation of ossicular prosthesis was accounted for another 2 cases, the hearing was improved after the ossicular orientation adjustment. One patient with lateral healing of TM and failed hearing recovery was corrected by a second operation. One case of tympanosclerosis underwent stapes release surgery, but hearing recovery still failed. One patient had recurrent postoperative cicatricial atresia of external auditory canal, and the patient was reluctant to undergo reoperation. Postoperative delayed facial paralysis occurred in 1 case, and the facial paralysis recovered recovered after conservative treatments. Conclusion:Eardrum patch and eustachian tube autoflation are simple and effective early outpatient treatment for patient with poor recovery. For those who failed with conservative treatments such as eardrum patch or eustachian tube and poor hearing recovery, the second surgical exploration is safe and effective. Regular follow up after endoscopic middle ear surgery is necessary for the managements of poor recovery.
Humans ; Ear, Middle/surgery* ; Female ; Male ; Endoscopy/methods* ; Adult ; Middle Aged ; Tympanic Membrane/surgery* ; Treatment Outcome ; Hearing Loss/surgery* ; Otologic Surgical Procedures/methods* ; Otitis Media/surgery* ; Eustachian Tube/surgery*

ObjectiveTo investigate the efficacy of ovarian tissue cryopreservation and autologous transplantation in preserving fertility and ovarian endocrine function in patients with cervical cancer. MethodsA 26-year-old patient with stage ⅡA1 cervical cancer underwent ovarian tissue harvesting and cryopreservation during cancer surgery. Following complete remission of the cancer， autologous ovarian tissue transplantation was performed. Follow-up monitoring included assessment of menopausal symptoms， hormone levels， and follicular development. ResultsSix months after transplantation， follicle-stimulating hormone levels decreased to 6.60 U/L， and estradiol levels increased from ＜10.00 ng/L to 89.00 ng/L. At 10 months after transplantation， ultrasound monitoring confirmed follicular development and physiological ovulation in the transplanted ovarian tissue. By 15 months after transplantation， follicle-stimulating hormone levels remained stable at 7.24 U/L， and estradiol levels further increased to 368.00 ng/L. Over 2 years after transplantation， the patient successfully gave birth to a healthy baby through assisted reproductive technology. ConclusionThe restoration of endocrine and ovulation functions in the transplanted cryopreserved ovarian tissue， followed by successful pregnancy， demonstrates the clinical success of ovarian tissue transplantation.

Objective:To investigate the effect of semaglutide on the metabolomics of obese patients with type 2 diabetes mellitus (T2DM) complicated by metabolic-associated fatty liver disease (MAFLD).Methods:A prospective non-randomized controlled study was conducted. Obese patients with T2DM complicated by MAFLD who attended the Department of Endocrinology of Shijiazhuang People′s Hospital from October 2022 to June 2023 were selected as the semaglutide group, and healthy individuals from the physical examination center were selected as the control group. Clinical data of both groups were collected. The semaglutide group was subcutaneously injected with semaglutide following a basic hypoglycemic regimen (starting dose of 0.25 mg once a week, which was changed to 0.5 mg once a week after 1 week for 12 weeks). Liquid chromatography-tandem mass spectrometry was used for qualitative and quantitative analyses of plasma metabolites, and multivariate analysis methods were used to analyze the metabolomics data.Results:In total, 69 patients in the semaglutide group completed the treatment, with 49 males (71%) and a median age of 46 (36, 54) years, and the healthy control group consisted of 100 individuals, with 38 males (38%) and a median age of 40 (35, 45) years. The body mass index and levels of fasting blood glucose, alanine aminotransferase, and interleukin-6 (IL-6) in the semaglutide group before treatment were significantly higher than those in the control group (all P<0.001). The body mass index [23.65 (22.33, 24.45) vs. 28.72 (27.50, 32.07) kg/m 2], liver stiffness measurement [1.61 (0.91, 2.00) vs. 5.78 (5.51, 6.10) kPa], and homeostasis model assessment of insulin resistance index [5.10 (2.90, 7.95) vs. 9.00 (6.25, 11.80)] in the semaglutide group were significantly lower after treatment than before treatment (all P<0.001), and the blood glucose, blood lipid, liver function indicator, and IL-6 levels all significantly decreased after treatment. Metabolomics analysis revealed that there were 219 differential metabolites (131 up-regulated and 88 down-regulated) between the semaglutide group ( n=27) before treatment and the control group ( n=12), with glycerophospholipids and free fatty acids being significantly up-regulated. The semaglutide group showed 203 differential metabolites (121 up-regulated and 82 down-regulated) after treatment compared with before, with significant down-regulation of long-chain fatty acids and significant up-regulation of metabolites including carnitines, branched-chain amino acids, and taurine. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the differential metabolites identified before and after semaglutide treatment were involved in several signaling pathways, such as biosynthesis of unsaturated fatty acids, linoleic acid metabolism, aldosterone synthesis and secretion, and the mTOR signaling pathway, etc. Conclusion:Semaglutide alters the serum metabolite levels in obese patients with T2DM complicated by MAFLD.

Objective To investigate the relationship between serum trimethylamine oxide(TMAO),neurofilament light chain protein(NfL),peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α)expression levels and short-term prognosis in aneurysmal subarachnoid hemorrhage(aSAH)patients.Method A total of 125 aSAH patients(aSAH group)and 125 healthy volunteers in the same period(control group)who were admitted in heji hospital affiliated to Changzhi Medical College from March 2020 to June 2023 were selected.The serum expression levels of TMAO,NfL and PGC-1α were compared between control group and aSAH group.The aSAH patients were followed up for 6 months after discharge.Their prognosis were evaluated using Glasgow Outcome Scale(GOS)and they were further divided into good prognosis and poor prognosis groups according to the GOS results.The serum expression levels of TMAO,NfL and PGC-1α were compared between the two groups.The poor prognosis influencing factors were analyzed by multivariate Logistic regression analysis,the serum TMAO,NfL and PGC-1α value in predicting poor prognosis were analyzed by receiver operating characteristic(ROC)curve.Result The expression levels of serum TMAO and PGC-1 α in the aSAH group were(2.63±0.36)μmol/L and(0.51±0.13)ng/mL,respectively,which were lower than those in the control group(3.18±0.57)μmol/L and(0.81±0.16)ng/mL(P<0.05).The expression level of serum NfL was significantly higher in the aSAH group(64.48±14.35 pg/mL)than in the control group(28.36±8.82 pg/mL)(P<0.05).Compared with the good prognosis group whose serum levels of TMAO and PGC-1 α were(2.80±0.80)μmol/L and(0.58±0.16)ng/mL,respectively,the poor prognosis group had significantly lower serum TMAO[(2.29±0.63)μmol/L]and PGC-1 α[(0.36±0.12)ng/mL](P<0.05).In contrast,poor prognosis group had a significantly higher level of NfL(76.70±15.61)pg/mL compared to good prognosis group(58.52±10.52)pg/mL(P<0.05).The proportion of patients with hypertension,patients with diabetes,patients with large or giant aneurysms,patients with Hunt Hess grade Ⅲ-Ⅳ,patients with onset to hospital time>12 h,and the level of C-reactive protein(CRP)were higher in the poor prognosis group than in the good prognosis group(P<0.05).Hunt Hess grade Ⅲ-Ⅳ,elevated serum CRP and NfL were independent risk factors for poor prognosis in aSAH patients(P<0.05),while elevated TMAO and PGC-1 α were protective factors(P<0.05).The area under the curve(AUC)of serum TMAO,NfL,PGC-1 α,and their combined prediction of poor prognosis in aSAH patients were 0.726,0.830,0.862,and 0.956,respectively.The AUC of the combined detection was greater than that of each indicator detected separately.Conclusion Serum TMAO and PGC-1α are lowly expressed in aSAH patients,and serum NfL is highly expressed,which are related to the occurrence of short-term poor prognosis,the combined detection of the three indicators has a high predictive value for short-term poor prognosis in aSAH patients.

BACKGROUND:Existing studies have made significant progress in PIWI-interacting RNAs(piRNAs)against osteoporosis,but the specific targets and related mechanisms by which piRNAs exert their functions remain to be explored.OBJECTIVE:To investigate the effects and downstream mechanisms of piR-7472 on the differentiation of mouse osteoblasts(MC3T3-E1 cells).METHODS:(1)Twelve C57/BL6J mice were randomly divided into a sham-operated and an ovariectomized group,with six mice in each group.Changes in bone mass and the expression of piR-7472 were detected using Micro-CT and RT-qPCR,respectively,at 8 weeks after surgery.(2)MC3T3-E1 cells were divided into NC mimics group,piR-7472 mimics group,NC inhibitor group,and piR-7472 inhibitor group.The mRNA expression of piR-7472,osteopontin,type I collagen,Runt-related transcription factor 2,and potassium voltage-gated channel modifier subfamily F member 1 were detected by RT-qPCR after 7 days of osteogenic induction.The protein expression of osteopontin,Runt-related transcription factor 2,bone morphogenetic protein 2,and potassium voltage-gated channel modifier subfamily F member 1(KCNF1)was detected using western blot assay.The expression of alkaline phosphatase was detected by alkaline phosphatase staining after 14 days of osteogenic induction,and the number of mineralized nodules was detected by alizarin red staining after 21 days of induction.Whether piR-7472 could bind to KCNF1 was observed by the dual luciferase reporter gene assay.RESULTS AND CONCLUSION:(1)Bone mineral density,bone volume fraction,bone trabecular thickness,bone trabecular number were significantly decreased and bone trabecular separation was significantly increased in ovariectomized mice,and piR-7472 in bone tissue was significantly down-regulated in osteoporotic mice.(2)Compared with the NC group,the mRNA expression of osteopontin,type I collagen,and Runt-related transcription factor 2 were significantly increased,the protein expression of osteopontin,Runt-related transcription factor 2,and bone morphogenetic protein 2 were significantly elevated,and the levels of mineralized deposition and alkaline phosphatase were increased in the piR-7472 mimics group.Compared with the NC inhibitor group,the mRNA expression of osteopontin,type I collagen,and Runt-related transcription factor 2 was significantly downregulated,the protein expression of osteopontin,Runt-related transcription factor 2,and bone morphogenetic protein 2 were significantly decreased,and the levels of mineralized deposition and alkaline phosphatase were reduced in the piR-7472 inhibitor group.(3)piR-7472 was found to interact with the potassium voltage-gated channel modifier subfamily F member 1 as predicted by the miRanda database.The dual luciferase reporter gene assay revealed that piR-7472 mimics could bind to and promote the expression of KCNF1.To conclude,piR-7472 can promote osteogenic differentiation of osteogenic precursor cells MC3T3-E1,and its mechanism of action may be achieved by promoting the expression of KCNF1.

Aim To investigate the correlation between serum solube growth stimulation expressed gene 2 protein(sST2)and early reperfusion arrhythmia(ERA)after emergency percutaneous coronary intervention(PCI)in patients with acute ST-segment elevation myocardial infarction(STEMI).Methods A total of 202 STEMI patients who under-went emergency PCI from November 2020 to August 2022 in the Cardiac Center of Liaoning Provincial People's Hospital were divided into two groups based on the occurrence of ERA within 48 hours after PCI:ERA group and non-ERA group.Serum sST2 level and clinical data were compared between the groups.Univariable and multivariable Logistic regression analysis were used to explore the association between serum sST2 level and ERA occurrence,and restricted cubic spline model was applied to identify independent risk factors for ERA.Results There were 83(41.1％)patients experi-enced ERA within 48 hours after PCI.Compared with the non-ERA group,the patients in ERA group had shorter time from chest pain to reperfusion and higher serum sST2 level(P＜0.001).Multivariate Logistic regression analysis showed that for STEMI patients,elevated serum sST2 level(sST2≥45.03 μg/L),early reperfusion time(chest pain to successful reperfusion time≤5.23 h),high thrombosis burden,and right coronary artery as the infarct related artery(IRA)were in-dependent risk factors for ERA after emergency PCI.The restricted cubic spline model suggested that the serum sST2 lev-el of STEMI patients was nonlinearly correlated with the risk of ERA after PCI(P＜0.01),and the cutoff point was 45.12 μg/L.ROC curve analysis showed that the area under the ROC curve of serum sST2 level in predicting ERA oc-currence after PCI was 0.827(95％CI:0.771～0.883).Conclusion The high serum sST2 level before PCI is an independent risk factor for ERA occurrence after PCI in patients with STEMI.When serum sST2＞45.12 μg/L,its level is positively correlated with the risk of ERA.

Objective:To investigate the feasibility of flight fatigue being detected via photoplethysmography (PPG) and regional cerebral oxygen saturation (rScO 2) in order to address the challenges posed by flight fatigue during prolonged or multiple consecutive flights. Methods:A total of 16 healthy male volunteers were enrolled. A wireless cerebral oximetry monitor headband was employed to collect PPG and rScO 2 data from the forehead while a multi-lead physiological data acquisition system was used concurrently to record three-lead electrocardiograms (ECGs). After 18 h of sleep deprivation, each volunteer performed a flight-simulating task, which was divided into 4 stages: the baseline period (T1), relaxation period (T2), early fatigue period (T3) and severe fatigue period (T4). Five-minute data was collected from each stage for analysis using AcqKnowledge 6.0. Heart rate (HR) and 3 HR variability (HRV) metrics, namely standard deviation of NN intervals (SDNN), root mean square of successive differences (RMSSD), and low frequency to high frequency power ratio (LF/HF), were computed independently from both ECG and PPG traces. The mean rScO 2 value for each stage was used to represent the cerebral oxygen saturation during that stage. The intra-class correlation coefficient (ICC) was employed to assess the consistency of the measurements, and the differences in HR and HRV indicators of the volunteers in the 4 stages of the experiment were analyzed. Results:The HR measured by ECG and PPG was highly consistent across the 4 stages among the 14 volunteers ( ICC=0.951, 0.963, 0.962, 0.963, P=0.013, 0.011, 0.021, 0.015), so were SDNN, RMSSD and LF/HF values ( ICC=0.935-0.983, all P<0.05). HR values calculated with either method showed significant differences across the 4 stages in the 14 volunteers ( F=21.63, 20.52, P=0.007, 0.008). HR gradually declined from T1 to T4, and was significantly lower at T4 than at T1 ( P=0.011, 0.009). There were significant differences in SDNN ( F=22.31, 24.26, P=0.006, 0.003), RMSSD ( F=22.30, 22.26, P=0.006, 0.006), and LF/HF ( F=20.37, 25.13, P=0.009, 0.002) across the 4 stages among the 14 volunteers. SDNN and RMSSD kept increasing as fatigue was intensified, while LF/HF decreased correspondingly. Statistically significant differences were found in SDNN, RMSSD and LF/HF values between T4 and T1 (all P<0.01). rScO 2 measured during the flight-simulating trial also differed significantly across the 4 stages ( F=21.39, P=0.007). rScO? at both T3 and T4 was significantly lower than at T1 ( P=0.009, 0.007). Conclusions:PPG can replace ECG for monitoring HR and HRV indicators under flight fatigue. Furthermore, the combination of PPG with rScO 2 monitoring allows for earlier detection of flight fatigue. This study is expected to offer a user-friendly and non-invasive approach to management of pilot fatigue.