Acupuncture, a therapeutic treatment defined as the insertion of needles into the body at specific points (ie, acupoints), has growing in popularity world-wide to treat various diseases effectively, especially acute and chronic pain. In parallel, interest in the physiological mechanisms underlying acupuncture analgesia, particularly the neural mechanisms have been increasing. Over the past decades, our understanding of how the central nervous system and peripheral nervous system process signals induced by acupuncture has developed rapidly by using electrophysiological methods. However, with the development of neuroscience, electrophysiology is being challenged by calcium imaging in view field, neuron population and visualization in vivo. Owing to the outstanding spatial resolution, the novel imaging approaches provide opportunities to enrich our knowledge about the neurophysiological mechanisms of acupuncture analgesia at subcellular, cellular, and circuit levels in combination with new labeling, genetic and circuit tracing techniques. Therefore, this review will introduce the principle and the method of calcium imaging applied to acupuncture research. We will also review the current findings in pain research using calcium imaging from in vitro to in vivo experiments and discuss the potential methodological considerations in studying acupuncture analgesia.
Calcium ; Acupuncture Therapy ; Acupuncture ; Acupuncture Analgesia/methods* ; Acupuncture Points ; Technology

Aim To explore the effects of Lycium berry seed oil on Nrf2/ARE pathway and oxidative damage in testis of subacute aging rats. Methods Fifty out of 60 male SD rats, aged 8 weeks, were subcutaneously injected with 125 mg • kg"D-galactosidase in the neck for 8 weeks to establish a subacute senescent rat model. The presence of senescent cells was observed using P-galactosidase ((3-gal), while testicular morphology was examined using HE staining. Serum levels of testosterone (testosterone, T), follicle-stimulating hormone ( follicle stimulating hormone, FSH ) , luteinizing hormone ( luteinizing hormone, LH ) , superoxide dis-mutase ( superoxide dismutase, SOD ) , glutathione ( glutathione, GSH) and malondialdehyde ( malondial-dehyde, MDA) were measured through ELISA, and the expressions of factors related to aging, oxidative damage, and the Nrf2/ARE pathway were assessed via immunohistochemical analysis and Western blotting. Results After successfully identifying the model, the morphology of the testis was improved and the intervention of Lycium seed oil led to a down-regulation in the expression of [3-gal and -yH2AX. The serum levels of SOD, GSH, T, and FSH increased while MDA and LH decreased (P 0. 05) . Additionally, there was an up-regulated expression of Nrf2, GCLC, NQOl, and SOD2 proteins in testicular tissue ( P 0. 05 ) and nuclear expression of Nrf2 in sertoli cells. Conclusion Lycium barbarum seed oil may reduce oxidative damage in testes of subacute senescent rats by activating the Nrf2/ARE signaling pathway.

AIM To explore the laws in accumulation of dry matter and medicinal ingredients of Isatis indigotica Fort.in Longdong area,and determine the best harvest time to obtain the high-quality and high-yield medicinal plant.METHODS The I.indigotica had its agronomic traits investigated regularly;the accumulation laws of its uridine,guanosine,(R,S)-geichan and adenosine measured by HPLC;and its comprehensive utilization value in different periods evaluated by grey correlation method.RESULTS There existed significant variations among the plant height,plant width,leaf length,leaf width,number of leaves,leaf weight,root length,root diameter,root weight,and the yields of uridine,guanosine,(R,S)-goitrin and adenosine at different growth stages,whose peak values appeared in different period.Additionally,there were also obvious differences in the aforementioned indexes levels among different strains.The highest weighted correlation degree of the comprehensive utilization value of Longde and 2008-6-2 on October 9,and of 2008-11 and 2016-2 on October 24.On the whole,the 2008-6-2 and 2016-2 were superior to 2008-11 and Longde.CONCLUSION It is suggested that the best harvest time of I.indigotica in Longdong area is in mid to late October.

Objective To investigate the effect of sophoridine on lipopolysaccharide(LPS)induced pulmonary alveolar epithelial cell damage by regulating the high-mobility group box 1(HMGB1)-receptor for advanced glycation end product(RAGE)signal pathway.Methods A549 and HAEC Ⅱ cells cultured in vitro were randomly divided into 5 groups:control group,LPS group,LPS+sophoridine group,LPS+empty group and LPS+sophoridine+HMGB1 overexpression group.Except for the control group,cells in other groups were induced by LPS to establish the injury models and treated with sophoridine and plasmids,respectively.Cell counting kit-8(CCK-8)method and terminal-deoxynucleoitidyl transferase mediated nick end labeling staining were used to detect cell viability and apoptosis rate in each group;Western blotting was used to detect the expression of HMGB1 and RAGE proteins.Results The A549 cell viability of control group,LPS group,LPS+sophoridine group,LPS+empty group and LPS+sophoridine+HMGB1 overexpression group were(100.00±0.00)％,(56.73±8.31)％,(90.02±11.24)％,(53.26±9.15)％and(60.84±8.13)％;the cell viability of HAEC Ⅱ were(100.00±0.00)％,(50.86±7.56)％,(93.05±12.38)％,(54.10±8.42)％and(54.43±6.14)％;the apoptosis rates of A549 cells were(2.11±0.65)％,(42.46±5.20)％,(4.01±1.31)％,(44.74±4.93)％and(39.75±4.86)％;the apoptosis rates of HAEC Ⅱ cells were(2.30±0.72)％,(48.14±4.87)％,(3.83±1.23)％,(45.72±5.14)％and(44.81±5.25)％;the HMGB1 protein levels in A549 cells were 0.16±0.02,0.87±0.13,0.19±0.04,0.89±0.11 and 0.84±0.12;RAGE protein levels were 0.19±0.03,0.94±0.16,0.21±0.04,0.96±0.15 and 0.90±0.17;the HMGB1 protein levels in HAEC Ⅱ cells were 0.13±0.04,0.79±0.10,0.15±0.04,0.80±0.14 and 0.75±0.12;RAGE protein levels were 0.28±0.07,1.08±0.19,0.31±0.06,1.10±0.21 and 1.04±0.15.The above indexes:there were statistically significant differences between LPS group and control group,LPS+sophoridine group and LPS group,LPS+sophoridine+HMGB1 overexpression group and LPS+sophoridine group(all P＜0.05).Conclusion Sophoritine can clear reactive oxygen species,reduce the production of inflammatory factors,enhance antioxidant enzyme activity,and inhibit LPS induced inflammation and oxidative stress in alveolar epithelial cells by reducing HMGB1-RAGE signal activity,ultimately reducing cell damage.

Objective To explore the regulatory mechanism of Macelignan on oxidative stress-mediated neuronal injury in autophagy and apoptosis.Methods Murine hippocampal neuronal HT22 cells were treated with 2.5 mmol·L-1 glutamic acid(Glu)to establish an oxidative stress cell model.The cells were divided into normal group(normal cultured cells),model group(2.5 mmol·L-1 Glu)and experimental-L,-M,-H groups(2.5,5,10 μmol·L-1Macelignan treatment),inhibitor group(2.5 mmol·L-1 Glu+10 μmol·L-1 Macelignan+10 μmol·L-1 LY294002).Aoptosis rate was detected by flow cytometry;the protein expression level of autophagy-related protein LC3B(LC3B),anti-SQSTM1/p62(p62),p21,B-cell lymphoma-2(Bcl-2)and Bcl-2 associated X protein(Bax)was detected by Western blot.Results The apoptosis rates in the normal group,model group and experimental-L,-M,-H groups were(4.58±1.25)％,(8.75±0.55)％,(6.30±1.71)％,(5.97±2.27)％and(5.49±1.71)％.The difference between model group and normal group was statistically significant(P＜0.01).The difference between experimental-L,-M,-H groups and model group was statistically significant(all P＜0.01).The levels of LC3B in normal group,model group,experimental-L,experimental-M,experimental-H groups and inhibitor group were 0.28±0.02,0.74±0.02,1.02±0.04,0.70±0.03,0.26±0.02 and 0.21±0.01;p62 levels were 0.49±0.08,0.33±0.03,0.50±0.07,0.59±0.01,0.64±0.13 and 0.65±0.06;p21 levels were 0.87±0.02,1.18±0.03,0.98±0.03,0.88±0.03,0.72±0.06 and 0.81±0.02;Bcl-2/Bax levels were 1.74±0.23,1.11±0.10,1.38±0.05,1.66±0.26,1.58±0.29 and 1.53±0.09,respectively.The differences between model group and normal group,between model group and experimental-H group,between model group and inhibitor group,were also statistically significant(all P＜0.01).Conclusion Macelignan can reduce the damage of hippocampal neurons induced by glutamate acid by regulating the process of autophagy and apoptosis,and has obvious neuroprotective effect.

Objective To investigate the inhibitory effect of Ginsenosides Rg1(GS-Rg1)on the proliferation and metastasis of tongue squamous cell carcinoma(TSCC),and its related mechanisms of action.Methods TSCC cells were treated with GS-Rg1 at concentrations of 1.25,2.5,5.0 and 10.0 μmol·L-1 for 48 hours.The proliferation ability of cells at different concentrations was measured by cell counting kit-8(CCK-8)experiment,and the IC5ovalue of GS-Rg1 at CAL-27 for 48 hours was calculated.TSCC cells CAL-27 were divided into control group and GS-Rg1 group.The control group and GS-Rg1 group were treated with 0.9％NaCl and IC50concentration of GS-Rg1 for 48 hours,respectively.The cell cycle distribution of each group was detected by flow cytometry,and the cell metastasis ability of each group was detected by Transwell experiment.Construct TOP/FOP Flash plasmid,transfect control group and GS-Rg1 group,and detect the effect of GS-Rg1 effect on wnt/β-catenin signaling pathway activity in TSCC cell CAL-27 using luciferase assay.Using wnt/β-catenin pathway inhibitor XAV939 treated GS-Rg1 group cells(XAV939+GS-Rg1 group),and wnt/β-catenin pathway activator HLY78 was used to treat GS-Rg1 group cells(HLY78+GS-Rg1 group)and detect changes of wnt/β-catenin signaling pathway activity,the cell proliferation ability,cell cycle distribution,and metastasis ability in XAV939+GS-Rg1 group,HLY78+GS-Rg1 group and GS-Rg1 group.The expression of wnt/β-catenin signaling pathway related proteins β-catenin,and its downstream cell cycle related proteins cellular myelocytomatosis oncogene(cMYC),Cyclin dependent kinase 4(CDK4),andcyclinD1,as well as metastasis related proteins E-cadherin,N-cadherin and matrix metalloproteinase 2(MMP-2)were detected by Western blotting in each group of cells.Results GS-Rg1 significantly inhibited the proliferation ability of TSCC cells CAL-27(P＜0.05),and the IC50value of GS-Rg1 on CAL-27 was(5.46±1.58)μmol·L-1.The ratio of GO/G1 phase cells in the control group and GS-Rg1 group were(60.65±2.16)％and(71.20±2.38)％,respectively;the number of cell transmembrane penetration were 87.33±7.51 and 50.67±3.21,respectively;the luciferase activity were 1.00±0.02 and 0.35±0.06,respectively.Compared with the control group,the GS-Rg1 group showed cell cycle arrest in GO/G1 phase,decreased cell metastasis ability,and the activity of wnt/β-catenin signaling pathway decreased(P＜0.05,P＜0.01).Compared with the GS-Rg1 group,the activity of the wnt/β-catenin signaling pathway was decreased,cell proliferation ability and metastasis ability was decreased(P＜0.05),while the number of GO/G1 phase cells was increased(P＜0.05),the expression of β-catenin,cMYC,CDK4,cyclinD1,E-cadherin and MMP-2 proteins were decreased(P＜0.05),while the expression of N-cadherin protein increased in XAV939+GS-Rg1 group cells.However,the result were opposite in the HLY78+GS-Rg1 group of cells.Conclusion GS-Rg1 downregulates wnt/β-catenin signaling pathway inhibits the proliferation and metastasis ability of TSCC cells.

Objective:To analyze the clinical features and risk factors of chest tightness variant asthma (CTVA) in children, so as to provide basis for the prevention and management of the disease.Methods:A cross-sectional study was conducted to analyze 178 children aged 6-17 years old who were admitted to the Department of Allergy, Capital Institute of Pediatrics Affiliated Children′s Hospital from January 2021 to January 2023 due to chest tightness. The age was 8.83(7.50, 11.58) years old, with 89 males (50%) and 89 females (50%). According to the diagnosis of CTVA, 130 cases were divided into CTVA group and 48 non-CTVA cases were divided into control group. Demographic data, personal history, family history, clinical features, auxiliary examination results and other data were collected. The clinical characteristics, allergens, FeNO level and pulmonary function parameters of the two groups were analyzed. Logistic regression analysis was used to explore the risk factors of the disease.Results:The proportion of school-age children (6-11 years old) in CTVA group was higher than that of adolescent children (≥12 years old) [(113/130,86.9%) vs (26/48,54.2%), Z=21.985, P<0.01]. The proportion of CTVA combined with eczema [(74/130,56.9%) vs (19/48,39.6%), χ2=4.225, P<0.05] and rhinitis symptoms [(98/130,75.4%) vs (27/48,56.2%), χ2=6.138, P<0.05] was higher. The positive rates of mold sensitization [(52/130,40.0%) vs (11/48,22.9%), χ2=4.474, P<0.05] and multiple sensitization [(71/130,54.6%) vs (18/48,37.5%), χ2=4.108, P<0.05] in inhaled allergens were significantly higher than those of control group. The proportion of elevated FeNO (>20 ppb) in CTVA children was 20.8% (27/130), which was significantly higher than that in control group 4.2%(2/48)( χ2=7.086 ,P<0.01). There were no statistical differences in spirometry parameters FEV 1 and FVC between CTVA group and control group ( P both>0.05). FEV 1/FVC, PEF, FEF 25, FEF 50, FEF 75 and MMEF were significantly lower than those in control group ( P all<0.05). Logistic regression analysis showed that rhinitis symptoms ( OR=2.351, 95% CI 1.105-5.002, P=0.026), multiple sensitizations ( OR=2.184, 95% CI 1.046-4.557, P=0.038), tIgE>60 kU/L( OR=3.080, 95% CI 1.239-7.654, P=0.015), FeNO>20 ppb ( OR=6.734, 95% CI 1.473-30.796, P=0.014) and small airway dysfunction ( OR=3.164, 95% CI 1.089-9.194, P=0.034) were risk factors for chest tightness variant asthma. FeNO combined with FEF 50 has the largest area under the curve ( Z=2.744, P<0.01) in diagnosing CTVA. Conclusion:CTVA is more common in school-age children than in adolescent children. Rhinitis symptoms, multiple sensitization, tIgE>60 kU/L, FeNO>20 ppb and small airway dysfunction are risk factors for chest tightness variant asthma. FeNO combined with small airway indexes can improve the diagnostic value of CTVA.

Objective To evaluate and compare the success rate and short-term clinical prognosis of transfemoral transcatheter aortic valve replacement(TF-TAVR)for patients with pure aortic regurgitation(PAR)of different annulus sizes.Methods This study is a single center retrospective study,selecting symptomatic PAR patients who received TF-TAVR treatment at Zhongshan Hospital Fudan University from September 2019 to September 2023.Based on preoperative CT results,all patients were divided into three groups:Group A(aortic annulus circumference＜80 mm),Group B(80 mm≤aortic annulus circumference＜85 mm),and Group C(aortic annulus circumference≥ 85 mm).The primary endpoint was success rate and 30d all-cause mortality,while the secondary endpoint was TAVR related complications.Results A total of 61 PAR patients were included in this study,including 27 in Group A,21 in Group B,and 13 in Group C.The overall success rate is 82.0％,and the 30 d all-cause mortality rate is 3.3％.The success rate of Group C patients was significantly lower(P=0.012),with higher rates of conversion to surgery and valve-in-valve implantation(P=0.022 and P=0.040).In terms of secondary endpoint events,there were no significant differences among the three groups in major bleeding events,major vascular complications,stroke,myocardial infarction,newly developed atrial fibrillation,implantation of new pacemakers,coronary artery occlusion,and postoperative moderate to severe perivalvular leakage(all P＞0.05).Conclusions The circumference of the aortic valve annulus is a key factor affecting the success rate of TF-TAVR in PAR,and PAR patients with an aortic valve annulus circumference less than 85mm may be more suitable for TF-TAVR.

Objective:To analyze the clinical features and risk factors of chest tightness variant asthma (CTVA) in children, so as to provide basis for the prevention and management of the disease.Methods:A cross-sectional study was conducted to analyze 178 children aged 6-17 years old who were admitted to the Department of Allergy, Capital Institute of Pediatrics Affiliated Children′s Hospital from January 2021 to January 2023 due to chest tightness. The age was 8.83(7.50, 11.58) years old, with 89 males (50%) and 89 females (50%). According to the diagnosis of CTVA, 130 cases were divided into CTVA group and 48 non-CTVA cases were divided into control group. Demographic data, personal history, family history, clinical features, auxiliary examination results and other data were collected. The clinical characteristics, allergens, FeNO level and pulmonary function parameters of the two groups were analyzed. Logistic regression analysis was used to explore the risk factors of the disease.Results:The proportion of school-age children (6-11 years old) in CTVA group was higher than that of adolescent children (≥12 years old) [(113/130,86.9%) vs (26/48,54.2%), Z=21.985, P<0.01]. The proportion of CTVA combined with eczema [(74/130,56.9%) vs (19/48,39.6%), χ2=4.225, P<0.05] and rhinitis symptoms [(98/130,75.4%) vs (27/48,56.2%), χ2=6.138, P<0.05] was higher. The positive rates of mold sensitization [(52/130,40.0%) vs (11/48,22.9%), χ2=4.474, P<0.05] and multiple sensitization [(71/130,54.6%) vs (18/48,37.5%), χ2=4.108, P<0.05] in inhaled allergens were significantly higher than those of control group. The proportion of elevated FeNO (>20 ppb) in CTVA children was 20.8% (27/130), which was significantly higher than that in control group 4.2%(2/48)( χ2=7.086 ,P<0.01). There were no statistical differences in spirometry parameters FEV 1 and FVC between CTVA group and control group ( P both>0.05). FEV 1/FVC, PEF, FEF 25, FEF 50, FEF 75 and MMEF were significantly lower than those in control group ( P all<0.05). Logistic regression analysis showed that rhinitis symptoms ( OR=2.351, 95% CI 1.105-5.002, P=0.026), multiple sensitizations ( OR=2.184, 95% CI 1.046-4.557, P=0.038), tIgE>60 kU/L( OR=3.080, 95% CI 1.239-7.654, P=0.015), FeNO>20 ppb ( OR=6.734, 95% CI 1.473-30.796, P=0.014) and small airway dysfunction ( OR=3.164, 95% CI 1.089-9.194, P=0.034) were risk factors for chest tightness variant asthma. FeNO combined with FEF 50 has the largest area under the curve ( Z=2.744, P<0.01) in diagnosing CTVA. Conclusion:CTVA is more common in school-age children than in adolescent children. Rhinitis symptoms, multiple sensitization, tIgE>60 kU/L, FeNO>20 ppb and small airway dysfunction are risk factors for chest tightness variant asthma. FeNO combined with small airway indexes can improve the diagnostic value of CTVA.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.