OBJECTIVE: To observe the effect of electroacupuncture at "Sishencong" (EX-HN1) and "Fengchi" (GB20) on hippocampal neuronal ferritinophagy mediated by the nuclear receptor coactivator 4 (NCOA4)/ferritin heavy chain 1 (FTH1) signaling pathway in vascular dementia (VD) rats, and to explore the potential mechanisms of electroacupuncture for VD. METHODS: A total of 60 male rats of SPF grade were randomly divided into a blank group (12 rats), a sham surgery group (12 rats) and a modeling group (36 rats). In the modeling group, the modified 4-vessel occlusion method was used to establish the VD model. The 24 successfully modeled rats were randomly divided into a model group and an electroacupuncture group, with 12 rats in each group. In the electroacupuncture group, electroacupuncture was applied at left and right "Sishencong" (EX-HN1), and bilateral "Fengchi" (GB20), with continuous wave, in frequency of 2 Hz and current intensity of 1 mA, 30 min a time, once daily for 21 consecutive days. The learning and memory abilities were assessed using the Morris water maze test before modeling, after modeling and after intervention, as well as the novel object recognition test after intervention. After intervention, the neuronal morphology in the hippocampus was observed by Nissl staining; the iron deposition was observed by Prussian blue staining; the reactive oxygen species (ROS) level was detected by dihydroethidium (DHE) fluorescence staining; the levels of iron, malondialdehyde (MDA) and superoxide dismutase (SOD) in the hippocampal tissue were measured by the colorimetric assay, TBA method, and WST-1 method, respectively; the positive expression of NCOA4, FTH1 and glutathione peroxidase 4 (GPX4) was detected by immunohistochemistry; the protein expression of NCOA4, FTH1, GPX4, and the ratio of microtubule-associated protein 1 light chain 3B (LC3B) Ⅱ/Ⅰ in the hippocampus were detected by Western blot. RESULTS: Compared with the sham surgery group, in the model group, the escape latency was prolonged, and the number of platform crossings reduced (P<0.01), the recognition index (RI) was decreased (P<0.01); the hippocampal neurons displayed a blurred laminar structure, disorganized cellular arrangement, and the number of Nissl bodies was decreased (P<0.01); the percentage of iron deposition area in the hippocampus was increased (P<0.01); in the hippocampus, the levels of ROS, iron, MDA, and the protein expression of NCOA4, as well as the LC3B Ⅱ/Ⅰ ratio were increased (P<0.01), the SOD level, and the protein expression of FTH1 and GPX4 were decreased (P<0.01). Compared with the model group, in the electroacupuncture group, the escape latency was shortened and the number of platform crossings was increased (P<0.01), the RI was increased (P<0.01); the hippocampal neurons exhibited more regular morphology, better-organized cellular structure, and the number of Nissl bodies was increased (P<0.05); the percentage of iron deposition area in the hippocampus reduced (P<0.01); in the hippocampus, the levels of ROS, iron, MDA, and the protein expression of NCOA4, as well as the LC3B Ⅱ/Ⅰ ratio were decreased (P<0.01, P<0.05), the SOD level, and the protein expression of FTH1 and GPX4 were increased (P<0.01). CONCLUSION Electroacupuncture at "Sishencong" (EX-HN1) and "Fengchi" (GB20) can improve learning and memory abilities in VD rats, and its mechanism may be associated with the regulation of the hippocampal NCOA4/FTH1 signaling pathway, inhibition of ferritinophagy, and alleviation of oxidative stress damage.
Animals ; Electroacupuncture ; Dementia, Vascular/genetics* ; Male ; Rats ; Signal Transduction ; Humans ; Memory ; Rats, Sprague-Dawley ; Nuclear Receptor Coactivators/genetics* ; Ferritins/genetics* ; Learning ; Hippocampus/metabolism* ; Acupuncture Points

Objective: To investigate the clinical efficacy of oral microscope-assisted microflap periodontal bone grafting in treating class Ⅱ furcation involvement in mandibular molars, and to provide clinical evidence for its treatment in furcation involvement. Methods: This study was reviewed and approved by the institutional ethics committee, and informed consent was obtained from all patients. Sixty mandibular molars with class II furcation involvement caused by periodontitis were enrolled in a randomized controlled clinical study, utilizing a random number table method. Patients were categorized into a control group (n=30) and an experimental group (n=30) based on the surgical procedure employed. The control group underwent periodontal flap surgery with an internal oblique incision and vertical incision; this procedure was performed without the aid of a microscope. Conversely, the experimental group underwent micro flap periodontal bone grafting surgery without vertical incision; an oral microscope was used for this procedure. Both groups were analyzed 6 months after surgery, and postoperative gingival recession (GR), probing depth (PD), bleeding index (BI), vertical bone height increase (VBHI), pain level, and complications were recorded. Results: Both groups showed improvement in PD and BI after 6 months compared to preoperative levels: the control group had a preoperative PD of (7.33 ± 1.72 mm) and a 6-month postoperative PD of (3.37 ± 0.96 mm), with statistically significant differences (P<0.001). The preoperative PD of the experimental group was (7.27 ± 1.57 mm), and the 6-month postoperative PD was (3.00 ± 0.69 mm), with statistically significant differences (P<0.001). The BI of the control group decreased from 3.03 ± 1.03 before surgery to 0.77 ± 0.82 at 6 months after surgery (P<0.001), while the BI of the experimental group decreased from 3.20 ± 1.09 before surgery to 0.73 ± 0.64 at 6 months after surgery (P<0.001), and the differences were statistically significant. The experimental group showed a significant improvement in GR (0.70 ± 0.59 mm) compared to preoperative GR (1.26 ± 0.94 mm) at 6 months after surgery (P=0.007), while the control group showed an increase in GR (1.37 ± 0.89 mm) at 6 months after surgery compared to preoperative GR (1.13 ± 0.97 mm), but the difference was not statistically significant (P=0.337). The inter group comparison results showed that there were no statistically significant differences in PD and BI between the two groups at 6 months after surgery (PD: P=0.096, BI: P=0.861); The GR of the experimental group was lower than that of the control group, and the difference was statistically significant (P=0.001). There was no statistically significant difference in postoperative VBHI between the two groups (P=0.128). The pain level scores of the experimental group were lower than those of the control group at 4 and 24 hours after surgery (P<0.001). None of the patients experienced complications. Conclusion Microflap periodontal bone grafting assisted by an oral microscope effectively improves the periodontal condition of patients with grade Ⅱ root bifurcation lesions of mandibular molars, and the bone grafting effect is good, with mild pain and good safety.

Hyperglycemia in pregnancy (HIP) exerts profound adverse effects on fetal development. While fetal cardiovascular abnormalities involve genetic and environmental factors, HIP serves as a critical environmental determinant that disrupts cardiovascular development through multiple pathways. These disruptions can lead to structural malformations, functional impairments, and long-term cardiovascular consequences in adulthood. Therefore, understanding HIP's impacts on offspring cardiovascular health and elucidating the underlying mechanisms are crucial.

Hyperglycemia in pregnancy (HIP) exerts profound adverse effects on fetal development. While fetal cardiovascular abnormalities involve genetic and environmental factors, HIP serves as a critical environmental determinant that disrupts cardiovascular development through multiple pathways. These disruptions can lead to structural malformations, functional impairments, and long-term cardiovascular consequences in adulthood. Therefore, understanding HIP's impacts on offspring cardiovascular health and elucidating the underlying mechanisms are crucial.

OBJECTIVE: To explore the mechanism of electroacupuncture (EA) at "Fengchi" (GB 20) and "Sishencong" (EX-HN 1) on learning and memory impairment in vascular dementia (VD) rats by observing the influences on the N-methyl-D-aspartate receptor (NMDAR)/cyclic adenosine monophosphate response element-binding protein (CREB)/brain-derived neurotrophic factor (BDNF) signaling pathway and the excitotoxicity induced by hippocampal calcium overload. METHODS: Thirty-two male SD rats of SPF grade were selected and randomized into a normal group (6 rats), a sham-operation group (6 rats) and an operation group (20 rats). VD model was established with the modified Pulsinelli's four-vessel occlusion (4-VO) method. Twelve rats after successfully modeled were assigned randomly into a model group and an EA group, 6 rats in each one. In the EA group, EA was delivered at bilateral "Fengchi" (GB 20) and "Sishencong" (EX-HN 1), with the continuous wave, the frequency of 2 Hz and the electric current of 1 mA. Stimulation intensity was adjusted depending on the slightly trembling of rat head. EA was given once daily, 30 min each time; and EA intervention was delivered for 21 days continuously. Using Morris water maze test, the learning and memory function was assessed. The neuronal morphology in the hippocampal CA1 was observed with HE staining; the level of glutamate (GLU) in serum and hippocampal tissue, as well as the activity of calcium pump (Ca²⁺-ATP) in the hippocampus were detected using colorimetric method. The protein expression of NMDAR, calmodulin-dependent protein kinase Ⅱ (CaMKⅡ), phosphorylated calmodulin-dependent protein kinase Ⅱ (p-CaMKⅡ), phosphorylated cyclic phosphoradenosine effector element binding proteins (p-CREB), CREB, and BDNF in the hippocampal CA1 was detected using immunohistochemistry. The protein expression of NMDAR, CREB, p-CREB and BDNF in the hippocampal tissue was detected using Western blot method. RESULTS: Compared to the sham-operation group, in the model group, the escape latency was prolonged and the platform crossing times of rats were reduced (P<0.01), the hippocampal neuron structure was damaged to different degrees, the structure in hippocampal CA1 was loosened, the arrangement disorganized, with clear grid-like structure; the neuronal morphology was irregular, pyknosis and even dissolution occurred, glial cells increased, blood capillary was dilated and the inflammatory cells were infiltrated and scattered. The level of GLU in the serum and hippocampal tissue and the protein expression of hippocampal NMDAR were elevated (P<0.01), the activity of Ca²⁺-ATP and the protein expression of CaMKⅡ, p-CaMKⅡ, CREB, p-CREB and BDNF were reduced (P<0.01, P<0.05); and the ratio of p-CaMKⅡ/CaMKⅡ and that of p-CREB/CREB were dropped (P<0.05). In comparison with the model group, in the EA group, the escape latency was shortened and the platform crossing times of rats rose (P<0.01), the arrangement was improved in the hippocampal CA1, the neuronal morphology was intact, the nucleoli were clear relatively and the pyknosis or dissolution were attenuated, the numbers of glial cells reduced relatively, the dilation of blood capillary was alleviated. The level of GLU in the serum and hippocampal tissue and the protein expression of NMDAR were reduced in the hippocampal tissue (P<0.01), the activity of Ca²⁺-ATP and the protein expression of CaMKⅡ, p-CaMKⅡ, CREB, p-CREB and BDNF were elevated (P<0.05, P<0.01); and the ratio of p-CaMKⅡ/CaMKⅡ and that of p-CREB/CREB increased (P<0.05). CONCLUSION EA at "Fengchi" (GB 20) and "Sishencong" (EX-HN 1) can attenuate learning and memory impairment in VD rats, which may be obtained by reducing GLU level in hippocampal tissue, inhibiting hippocampal excitotoxicity, mediating protein expression related to the NMDAR/CREB/BDNF signaling pathway, and maintaining neuronal survival and growth.
Electroacupuncture ; Male ; Animals ; Rats, Sprague-Dawley ; Learning ; Memory ; Signal Transduction ; Cyclic AMP Response Element-Binding Protein/metabolism* ; Memory Disorders/therapy* ; Brain-Derived Neurotrophic Factor/metabolism* ; Receptors, N-Methyl-D-Aspartate/metabolism* ; Dementia, Vascular/therapy*

Gelsemium elegans, a vine plant of Loganiaceae, has both medicinal and forage values. However, it is susceptible to low temperatures during growth. Exploring low temperature response genes is of great significance for cold resistance breeding of G. elegans. Ethylene response factors (ERFs) are the transcription factors of the AP2/ERF superfamily and play a crucial role in plant stress response. In this study, based on the unigene GeERF involved in the response to low temperature stress in the transcriptome of G. elegans, a full-length cDNA sequence of the transcription factor GeERF4B-1 was cloned from the leaves of G. elegans by reverse transcription-polymerase chain reaction (RT-PCR). Bioinformatics analysis showed that GeERF4B-1 had an open reading frame of 759 bp, encoding a protein composed of 252 amino acid residues and with a relative molecular weight of 27 kDa. The deduced protein was predicted to be an unstable, alkaline, and hydrophilic protein. The phylogenetic tree showed that GeERF4B-1 was in the same clade as the B-4 subfamily of the ERF family. The results of the subcellular localization experiment revealed that GeERF4B-1 was located in the nucleus. Real time quantitative PCR (RT-qPCR) analysis indicated that GeERF4B-1 was expressed in the root, stem, and leaf of G. elegans, with the highest expression level in the root. Compared with the control, the treatments with a low temperature (4 ℃), methyl jasmonate (MeJA), and abscisic acid (ABA) up-regulated the expression level of GeERF4B-1, which reached the peak at 24-48 h. This result revealed that GeERF4B-1 actively responded to low temperature, MeJA, and ABA stresses. However, both sodium chloride (NaCl) and drought treatments down-regulated the expression of GeERF4B-1. In addition, a prokaryotic expression vector of GeERF4B-1 was constructed, and a fusion protein of approximately 52 kDa was yielded after induced expression. The results of the plate stress assay showed that compared with the control, the prokaryotic strain expressing GeERF4B-1 demonstrated enhanced tolerance to low temperatures and sensitivity to salt and mannitol stresses. This study provides theoretical references and potential genetic resources for breeding G. elegans varieties with stress resistance.
Transcription Factors/metabolism* ; Plant Proteins/metabolism* ; Gelsemium/metabolism* ; Acetates/pharmacology* ; Gene Expression Regulation, Plant ; Phylogeny ; Cold Temperature ; Amino Acid Sequence ; Cyclopentanes/metabolism* ; Oxylipins/metabolism* ; Stress, Physiological/genetics* ; Abscisic Acid/metabolism* ; Cloning, Molecular

【Objective】 To monitor the effectiveness and accuracy of the blood transfusion compatibility test system by self-made weakly positive internal quality control products. 【Methods】 Red blood cells from DAT(-) healthy subjects were selected, and B/RhD(-)E(-) red blood cells were selected as tube 1. A/RhD(+ )E(+ ) was selected as tube 2 to prepare blood group quality control products according to the principle of blood group antigen compatibility, and red blood cell preservation solution and corresponding ABO blood group reagent antibody were added to make the agglutination intensity of microcolumn gel method in reverse blood typing reach a low positive value (1+ ). Tube 3 and tube 4 were prepared with five different preservation media: plasma, serum, antibody diluent, mixture of equal plasma and antibody diluent, and mixture of equal serum and antibody diluent, respectively. IgM anti-E antibody was added to tube 3, and IgG anti-D antibody was added to tube 4, so that the agglutination intensity of microcolumn gel method reached a low positive value (1+ ). 【Results】 Comparison between the 5 different preservation media showed that the preservation medium of antibody diluent was the most stable for weakly positive antibody (F=11.35, P<0.05), Agglutination intensity 1+ is assigned 5 points by AABB Technical Manual, and its score was 5.25±1.75 points. 【Conclusion】 The use of self-made weakly positive quality control products can improve the effectiveness, accuracy and sensitivity of the monitoring system, thus achieving internal quality control and ensuring the safety of clinical blood use.

Objective:To evaluate safety and efficacy of B-type suture method ileostomy.Methods:Clinical data from 204 patients undergoing laparoscopic low anterior resection combined with protective ileostomy was analysed. Patients were divided into B-type suture ileostomy group ( n=67) and traditional ileostomy group ( n=137). Results:compared with traditional ileostomy group, B-type suture ileostomy group showed statistically significant differences in total operation time [(164±26) min vs. (172±24) min, t=2.229, P=0.027], ileostomy time [(12.7±2.3) min vs. (14.8±2.2) min, t=-6.565, P<0.001], blood loss [(57±20) ml vs. (69±31) ml, t=-2.797, P=0.006], postoperative hospital stay [(10.2±1.9) d vs. (11.8±2.3) d, t=-4.851, P<0.001], specimen incision infection rate (0 vs. 5.1%, P=0.047), postoperative body pain [82 (79-84) vs. 78 (76-80), Z=-5.805, P<0.001], and ileostomy incorporation time [(46±11) min vs. (51±12) min, t=-2.540, P=0.012]. Conclusion:B-type suture ileostomy for prophylactic ileostomy in laparoscopic low anterior resection for rectal cancer is safe and feasible.

Objective To determine the reference red blood cells with weak agglutination intensity of low positive quali-ty control products by comparing RhD antigen expression intensity difference according to the serological results.Methods The RhD(+)red blood cells were detected by microcolumn gel method with 1 500 times diluted anti-D typing reagent.The samples with weak and strong RhD antigen expression intensity were selected as the reference red blood cells for weak agglu-tination intensity of low positive quality control products,and verification was performed.Results Ten RhD(+)red blood cells were detected with diluted anti-D typing reagent,of which 8 were 1+and 2 were±.Red blood cells with agglutination intensity of 1+were used as the benchmark to determine the maximum dilution ratio of anti-D typing reagent when their ag-glutination intensity was 1+.As the preparation standard of low positive quality control products,the agglutination intensity of red blood cells with low RhD antigen expression intensity was extremely weak±,which was difficult to ensure the stability of its control limit properties.Based on red blood cells with agglutination intensity of±,the maximum dilution ratio of anti-D typing reagent with agglutination intensity of 1+was re-determined as the preparation standard of low positive quality con-trol products,and the results met the requirements of quality control product setting.Conclusion Using red blood cells with low RhD antigen expression intensity as the benchmark to set the weak agglutination intensity of the low positive quality control products can avoid the loss of control due to the low target value.

OBJECTIVE To investigate the protective effect of paeoniflorin(PF) on cardiac dysfunction and myocardial cell injury induced by cisplatin(CDDP) in rats. METHODS SD male rats were randomly divided into control group, CPPD group, and CDDP PF+low-dose, high-dose group. PowerLab multifunctional recorder was used to detect the related indexes of cardiac function: the changes of left ventricular peak pressure(LVSP), left ventricular end-diastolic pressure(LVEDP) and left ventricular pressure change rate(±dp/dt). Serum levels of inflammatory factors TNF-α, IL-1β and IL-6 were measured in each group. Myocardial tissue was stained to observe the changes of tissue structure. H9c2 cardiomyocytes were divided into control group, CDDP group, PF group and CDDP+PF group. The activity of H9c2 cardiomyocytes was measured by CCK-8. The apoptosis of cardiomyocytes in each group was detected by flow cytometry. The expressions of MAPK signaling pathway related proteins p38, ERK, JNK and their phosphorylated proteins and apoptosis-related proteins Bax, Bcl-2, Casp3, Cl-casp3 were detected in cardiomyocytes by Western blotting. RESULTS Compared with the control group, LVSP and ±dp/dt decreased, LVEDP increased in rats of CDDP group(P<0.01). Compared with CDDP group, both CDDP+low-dose and high-dose PF pretreatment increased LVSP and ±dp/dt value(P<0.05 or P<0.01), decreased LVEDP(P<0.01), and could decrease the serum inflammatory factor TNF-α, IL-1β and IL-6(P<0.01). Cell level results showed that compared with control group, in CDDP group, the cell activity decreased, the apoptosis-related protein Bax, Cl-casp3 increased(P<0.01), expression of anti-apoptotic protein Bcl-2 decreased(P<0.01), and the expression of p38 and ERK phosphorylation also increased(P<0.01). Compared with CDDP group, PF could restore cell activit, down-regulate apoptosis-related protein Bax, Cl-casp3(P<0.05 or P<0.01), and increase anti-apoptotic protein Bcl-2 expression(P<0.01), inhibit MAPK pathway p38 and ERK phosphorylation expression(P<0.01). CONCLUSION PF can restore cardiac dysfunction and myocardial cell injury induced by cisplatin in rats, which may be related to inhibiting inflammation and apoptosis by regulating ERK/p38 MAPK signal expression.