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

Targeted protein degradation via nanoparticle-based universal strategy modifies nanoparticles with antibodies and ingeniously utilizes its cellular transport characteristics. This strategy achieved targeted degradation of extracellular proteins without complex design.Image 1.

Objective To explore the value of the ratio of soluble FmS-like tyrosine kinase-1(sFlt-1)to placental growth factor(PlGF)in predicting vitamin D deficiency in pregnant women with preeclampsia.Methods A total of 740 singleton pregnant women who underwent prenatal examination in Qijiang Health Center for Maternal and Child Care from January to November 2023 and were able to complete follow-up were selected as the research subjects.They were divided into the normal group(n=44)and the deficiency group(n=696)according to the vitamin D[1,25-(OH)2D3]level.Information such as age,BMI,pregnancy histo-ry,family history and medical history of each group was collected.5 mL of peripheral venous blood was drawn after fasting for 6-8 hours at 12-14 weeks of pregnancy to detect the levels of sFlt-1,PlGF and vitamin D in peripheral blood,and evaluated the predictive value of sFlt-1/PlGF for vitamin D deficiency in preeclampsia.Results There were no statistically significant differences in general conditions such as height,age,BMI,edu-cational level and occupation between the two groups(P>0.05).There were statistically significant differ-ences in the prenatal weight,vitamin D level,sFlt-1/PlGF and neonatal birth weight between the two groups(P<0.05).The results of univariate analysis showed that the prenatal weight,sFlt-1/PlGF,and neonatal birth weight were risk factors for vitamin D deficiency in patients with PE.The results of multivariate analysis showed that sFlt-1/PlGF and neonatal birth weight were factors for predicting vitamin D deficiency in patients with PE(P<0.05).All these variables are used to establish the nomogram prediction model.The receiver op-erating characteristic(ROC)curve showed that area under the curve(AUC)of the training group and the val-idation group was 0.66(95%CI:0.56-0.77)and 0.63(95%CI:0.49-0.76),respectively.Conclusion In pregnant women with preeclampsia,elevated sFlt-1/PlGF may be an early warning indicator for predicting vi-tamin D deficiency.

Objective To explore the value of peripheral blood monocyte subsets and their CD169 expression levels in the diagnosis of common respiratory viral infections and the immune mechanism of the infection in children.Methods A total of 125 children with re-spiratory viral infections(virus infection group),71 children with bacterial respiratory infections(bacteria infection group),and 35 healthy children(healthy control group)admitted to the Third Affiliated Hospital of Zhengzhou University from February 2022 to Octo-ber 2023 were included in this study.The counts of peripheral blood monocytes and lymphocytes were detected by hematologic analyzer.The proportions of lymphocyte subsets,monocyte subsets,and the expression levels of CD169 were detected by a flow cytometry.Mono-cyte subsets were classified into three types:classical monocytes(CM),intermediate monocytes(IM),and non-classical monocytes(NCM).ANOVA and Kruskal-Wallis H tests were used to screen for differentially expressed indicators among the three groups,and the receiver operator characteristic(ROC)curve was used to evaluate their diagnostic performance.Paired t-tests were used to analyze the differences in the indicators between the first day and the seventh day of hospitalization in the virus group.Spearman rank correlation a-nalysis was used to explore the relationship between CD 169 expression levels in monocyte subsets and the quantity of lymphocyte sub-sets.Results Statistically significant differences in the expression levels of CD169 in monocytes of CM,IM,and NCM types were ob-served in peripheral blood between the children's respiratory viral infection group and the healthy control group(all P＜0.01).In con-trast,no statistically significant differences in CD169 expression levels were found in monocytes of CM and IM type between the respira-tory viral infection group and the bacterial infection group(all P＞0.05).The differences in CD 169 expression levels among the groups were statistically significant in the NCM subset(P=0.008).The area under the ROC curve(AUC)of CD169 expression level in NCM type monocytes for diagnosing respiratory viral infections was 0.897(P＜0.01),with a cutoff point of ≥ 62.75％,sensitivity of 68.60％,and specificity of 91.80％.The quantity and proportion of CM,IM,and NCM type monocytes did not show statistically signif-icant differences between the respiratory viral infection group and the bacterial infection group(all P＞0.05).However,there were sig-nificant differences in the quantity and proportion of CM and IM type monocytes,as well as the proportion of NCM type monocytes,be-tween the virus infection group and the healthy control group(all P＜0.05).The proportions of CM,IM,and NCM type monocytes,the quantity of IM type monocytes,and the CD169 expression levels of monocyte subsets showed statistically significant differences between the first day and the seventh day after admission(all P＜0.05).There were no statistically significant differences in the quantity of CM and NCM type monocytes between the first day and the seventh day after admission(all P＞0.05).The expression levels of CD169 in peripheral blood CM,IM,and NCM type monocytes were negatively correlated with the number of CD8+T cells with correlation coeffi-cients of-0.65,-0.63,and-0.66,respectively(all P＜0.01).Conclusion The expression level of CD169 in NCM type monocytes could be used as an auxiliary diagnostic marker for the children with respiratory viral infection.The changes of monocyte subsets during respiratory viral infection course in children may involve that more CM type monocytes differentiate into IM type monocytes,and some IM type monocytes do not differentiate into NCM type monocytes in proportion.IM and NCM type monocytes may play important roles in the early stage of respiratory viral infections in children.CD169 serves as a marker of monocyte activation.

Objective To explore the value of peripheral blood monocyte subsets and their CD169 expression levels in the diagnosis of common respiratory viral infections and the immune mechanism of the infection in children.Methods A total of 125 children with re-spiratory viral infections(virus infection group),71 children with bacterial respiratory infections(bacteria infection group),and 35 healthy children(healthy control group)admitted to the Third Affiliated Hospital of Zhengzhou University from February 2022 to Octo-ber 2023 were included in this study.The counts of peripheral blood monocytes and lymphocytes were detected by hematologic analyzer.The proportions of lymphocyte subsets,monocyte subsets,and the expression levels of CD169 were detected by a flow cytometry.Mono-cyte subsets were classified into three types:classical monocytes(CM),intermediate monocytes(IM),and non-classical monocytes(NCM).ANOVA and Kruskal-Wallis H tests were used to screen for differentially expressed indicators among the three groups,and the receiver operator characteristic(ROC)curve was used to evaluate their diagnostic performance.Paired t-tests were used to analyze the differences in the indicators between the first day and the seventh day of hospitalization in the virus group.Spearman rank correlation a-nalysis was used to explore the relationship between CD 169 expression levels in monocyte subsets and the quantity of lymphocyte sub-sets.Results Statistically significant differences in the expression levels of CD169 in monocytes of CM,IM,and NCM types were ob-served in peripheral blood between the children's respiratory viral infection group and the healthy control group(all P＜0.01).In con-trast,no statistically significant differences in CD169 expression levels were found in monocytes of CM and IM type between the respira-tory viral infection group and the bacterial infection group(all P＞0.05).The differences in CD 169 expression levels among the groups were statistically significant in the NCM subset(P=0.008).The area under the ROC curve(AUC)of CD169 expression level in NCM type monocytes for diagnosing respiratory viral infections was 0.897(P＜0.01),with a cutoff point of ≥ 62.75％,sensitivity of 68.60％,and specificity of 91.80％.The quantity and proportion of CM,IM,and NCM type monocytes did not show statistically signif-icant differences between the respiratory viral infection group and the bacterial infection group(all P＞0.05).However,there were sig-nificant differences in the quantity and proportion of CM and IM type monocytes,as well as the proportion of NCM type monocytes,be-tween the virus infection group and the healthy control group(all P＜0.05).The proportions of CM,IM,and NCM type monocytes,the quantity of IM type monocytes,and the CD169 expression levels of monocyte subsets showed statistically significant differences between the first day and the seventh day after admission(all P＜0.05).There were no statistically significant differences in the quantity of CM and NCM type monocytes between the first day and the seventh day after admission(all P＞0.05).The expression levels of CD169 in peripheral blood CM,IM,and NCM type monocytes were negatively correlated with the number of CD8+T cells with correlation coeffi-cients of-0.65,-0.63,and-0.66,respectively(all P＜0.01).Conclusion The expression level of CD169 in NCM type monocytes could be used as an auxiliary diagnostic marker for the children with respiratory viral infection.The changes of monocyte subsets during respiratory viral infection course in children may involve that more CM type monocytes differentiate into IM type monocytes,and some IM type monocytes do not differentiate into NCM type monocytes in proportion.IM and NCM type monocytes may play important roles in the early stage of respiratory viral infections in children.CD169 serves as a marker of monocyte activation.

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*

Objective To invesitgate the epidemiological characteristics of common allergens in 10 664 patients with allergic diseases in Zhengzhou area.Methods A total of 10 664 patients visited our hospital and underwent serum allergen screening during January 2013 and August 2021 were selected,and their serum sIgE results were retrospectively analyzed.Results The total positive rate of sIgE to allergens in 10 664 patients was 69.82%.The positive rate of sIgE to inhalant allergens was significantly higher than that to in-gestive allergens(χ2=99.15,P＜0.01).The top three inhalant allergens were grass and tree combination,dust mite combination,and cockroach.The top three ingestive allergens were egg protein,milk,and seafood combination.The positive rate of sIgE to ingestive al-lergens in males was significantly higher than that in females(χ2=8.18,P＜0.01).The highest positive rate of sIgE to ingestive aller-gens was found in the early childhood period(χ2=125.92,P＜0.05).The highest positive rate of sIgE to inhalant allergens was found in the school-age and preschool periods(χ2=283.76,P＜0.01).The proportions of sIgE to cockroach and house dust mite showed a de-creasing trend year by year,while the proportions of sIgE to milk,peanut,lamb,and seafood combination showed an increasing trend year by year.Conclusion The top three inhalant and ingestive allergens in Zhengzhou area are grass and tree combination,dust mite,cockroach and egg protein,milk,seafood combination,respectively.In recent years,the allergies to milk,peanut,lamb,and seafood should be paid attention.

Vascular dementia(VD)is caused by cerebrovascular diseases,either hemorrhage or ischemic damage in the brain,with ischemia being the most common.In recent years,increasing efforts have been made to study the etiology,pathogenesis,and prevention of VD.The establishment of appropriate animal models to study the mechanism of VD and explore the efficacy of VD treatments has become an important issue in this research field.On the basis of conventional method,such as bilateral occlusion of common carotid arteries(2VO)and four-vessel occlusion,researchers have modified these method to improve stability with better reflection of the clinical manifestations of VD.This review summarizes these modified method and discusses possible cellular and molecular mechanisms and their advantages and disadvantages.

Objective:To summarize the morphological characteristics of diffuse large B-cell lymphoma (DLBCL) cells and investigate the prognosis value of the characteristics and the number of DLBCL cells in bone marrow.Methods:Retrospective study. We collected 79 cases newly diagnosed with DLBCL in the First Affiliated Hospital of Zhengzhou University from January 2020 to August 2022. 30 cases newly diagnosed without bone marrow involvement were selected as controls, whose mean age 58 years (30-82 years). The DLBCL cells were evaluated by the bone marrow smear, biopsy and flow cytometry separetely.The detection rate of DLBCL cells in the bone marrow was compared, to analyse the relationship between the morphological characteristics of DLBCL in the smear, clinical characteristics and flow cytometry parameters, and the prognostic value of DLBCL detected in the bone marrow smear and its quantity was analyzed. Logistic regression was used to analyze the correlation between the detection of DLBCL cells in bone marrow smears and the age, clinical stage, and the number of extraderules involved organs. Multivariate Cox regression was used to analyze the influence of DLBCL cells detection and its number on the prognosis of patients.Results:(1) The positive rates of DLBCL cells in bone marrow biopsy, bone marrow smear and flow cytometry were 4.86%, 5.14% and 9.27% respectively. (2) The morphological characteristics of 79 cases in bone marrow smears were described: more than 2 times the volume of the cell body of the lymphocyte, the shape was different, round or quasi-round or irregular shape, can be seen pseudopodia or protrusion; The volume of cytoplasm was moderate, vacuoles were visible, and a few perinuclear areas were visible. The nucleus were different in shape, round or quasi-round or irregularly shaped, with a majority of them having multiple nuclei and a few of them having delicate and loose chromatin. Most nucleoli were medium or large obviously, with a majority of them having 1-2 nucleoli and a few having more than 3.Sergiosomes and hemophagocytosis were observed in some DLBCL cases, tumor cell aggregation phenomenon was observed in a few DLBCL cases, occasionally pathological mitosis.(3) DLBCL cells in bone marrow smear was positively related to the age of patients, clinical stage and the number of extranodal organs involved(regression coefficient were 2.012, 2.754, 2.028, P<0.05);The volume of DLBCL cells in bone marrow smear was positively correlated with the ratio of CD4 and CD8(regression coefficient is 2.545, P<0.05);The vacuoles in cytoplasm and the pseudopod of tumor were both negative relationship with the quantity of CD38 expressed on DLBCL cells(regression coefficient was -2.465, -3.045, P<0.05); (4) DLBCL cells in bone marrow smear was an independent risk factor for PFS and OS( RR=7.059, P<0.05); RR=5.409, P<0.05). Conclusion:The appearace of DLBCL cells in bone marrow smear with prognosis, and could be used for clinical staging.

Necrosis is a form of cell death, which is related to various serious diseases such as cardiovascular disease, cancer, and neurodegeneration. Necrosis-avid agents (NAAs) selectively accumulated in the necrotic tissues can be used for imaging and/or therapy of related diseases. The aim of this study was to preliminarily investigate necrosis avidity of I-evans blue (I-EB) and its mechanism. The biodistribution of I-EB at 24 h after intravenous administration showed that the radioactivity ratio of necrotic to viable tissue was 3.41 in the liver and 11.82 in the muscle as determined by counting in model rats. Autoradiography and histological staining displayed preferential uptake of I-EB in necrotic tissues. nuclear extracts from necrotic cells exhibited 82.3% of the uptake in nuclei at 15 min, as well as 79.2% of the uptake at 2 h after I-EB incubation. The DNA binding study demonstrated that evans blue (EB) has strong binding affinity with calf-thymus DNA (CT-DNA) (=5.08×10 L/(mol/L)). Furthermore, the accumulation of I-EB in necrotic muscle was efficiently blocked by an excess amount of unlabeled EB. In conclusion, I-EB can not only detect necrosis by binding the DNA released from necrotic cells, but also image necrotic tissues generated from the disease clinically.