1.Mechanism of salvianolic acid B protecting H9C2 from OGD/R injury based on mitochondrial fission and fusion
Zi-xin LIU ; Gao-jie XIN ; Yue YOU ; Yuan-yuan CHEN ; Jia-ming GAO ; Ling-mei LI ; Hong-xu MENG ; Xiao HAN ; Lei LI ; Ye-hao ZHANG ; Jian-hua FU ; Jian-xun LIU
Acta Pharmaceutica Sinica 2024;59(2):374-381
		                        		
		                        			
		                        			 This study aims to investigate the effect of salvianolic acid B (Sal B), the active ingredient of Salvia miltiorrhiza, on H9C2 cardiomyocytes injured by oxygen and glucose deprivation/reperfusion (OGD/R) through regulating mitochondrial fission and fusion. The process of myocardial ischemia-reperfusion injury was simulated by establishing OGD/R model. The cell proliferation and cytotoxicity detection kit (cell counting kit-8, CCK-8) was used to detect cell viability; the kit method was used to detect intracellular reactive oxygen species (ROS), total glutathione (t-GSH), nitric oxide (NO) content, protein expression levels of mitochondrial fission and fusion, apoptosis-related detection by Western blot. Mitochondrial permeability transition pore (MPTP) detection kit and Hoechst 33342 fluorescence was used to observe the opening level of MPTP, and molecular docking technology was used to determine the molecular target of Sal B. The results showed that relative to control group, OGD/R injury reduced cell viability, increased the content of ROS, decreased the content of t-GSH and NO. Furthermore, OGD/R injury increased the protein expression levels of dynamin-related protein 1 (Drp1), mitofusions 2 (Mfn2), Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase 3), and decreased the protein expression levels of Mfn1, increased MPTP opening level. Compared with the OGD/R group, it was observed that Sal B had a protective effect at concentrations ranging from 6.25 to 100 μmol·L-1. Sal B decreased the content of ROS, increased the content of t-GSH and NO, and Western blot showed that Sal B decreased the protein expression levels of Drp1, Mfn2, Bax and caspase 3, increased the protein expression level of Mfn1, and decreased the opening level of MPTP. In summary, Sal B may inhibit the opening of MPTP, reduce cell apoptosis and reduce OGD/R damage in H9C2 cells by regulating the balance of oxidation and anti-oxidation, mitochondrial fission and fusion, thereby providing a scientific basis for the use of Sal B in the treatment of myocardial ischemia reperfusion injury. 
		                        		
		                        		
		                        		
		                        	
2.Hepatitis C virus infection:surveillance report from China Healthcare-as-sociated Infection Surveillance System in 2020
Xi-Mao WEN ; Nan REN ; Fu-Qin LI ; Rong ZHAN ; Xu FANG ; Qing-Lan MENG ; Huai YANG ; Wei-Guang LI ; Ding LIU ; Feng-Ling GUO ; Shu-Ming XIANYU ; Xiao-Quan LAI ; Chong-Jie PANG ; Xun HUANG ; An-Hua WU
Chinese Journal of Infection Control 2024;23(1):1-8
		                        		
		                        			
		                        			Objective To investigate the infection status and changing trend of hepatitis C virus(HCV)infection in hospitalized patients in medical institutions,and provide reference for formulating HCV infection prevention and control strategies.Methods HCV infection surveillance results from cross-sectional survey data reported to China Healthcare-associated Infection(HAI)Surveillance System in 2020 were summarized and analyzed,HCV positive was serum anti-HCV positive or HCV RNA positive,survey result was compared with the survey results from 2003.Results In 2020,1 071 368 inpatients in 1 573 hospitals were surveyed,738 535 of whom underwent HCV test,4 014 patients were infected with HCV,with a detection rate of 68.93%and a HCV positive rate of 0.54%.The positive rate of HCV in male and female patients were 0.60%and 0.48%,respectively,with a statistically sig-nificant difference(x2=47.18,P<0.001).The HCV positive rate in the 50-<60 age group was the highest(0.76%),followed by the 40-<50 age group(0.71%).Difference among all age groups was statistically signifi-cant(x2=696.74,P<0.001).In 2003,91 113 inpatients were surveyed.35 145 of whom underwent HCV test,resulting in a detection rate of 38.57%;775 patients were infected with HCV,with a positive rate of 2.21%.In 2020,HCV positive rates in hospitals of different scales were 0.46%-0.63%,with the highest in hospital with bed numbers ranging 600-899.Patients'HCV positive rates in hospitals of different scales was statistically signifi-cant(X2=35.34,P<0.001).In 2020,12 provinces/municipalities had over 10 000 patients underwent HCV-rela-ted test,and HCV positive rates ranged 0.19%-0.81%,with the highest rate from Hainan Province.HCV posi-tive rates in different departments were 0.06%-0.82%,with the lowest positive rate in the department of pedia-trics and the highest in the department of internal medicine.In 2003 and 2020,HCV positive rates in the depart-ment of infectious diseases were the highest,being 7.95%and 3.48%,respectively.Followed by departments of orthopedics(7.72%),gastroenterology(3.77%),nephrology(3.57%)and general intensive care unit(ICU,3.10%)in 2003,as well as departments of gastroenterology(1.35%),nephrology(1.18%),endocrinology(0.91%),and general intensive care unit(ICU,0.79%)in 2020.Conclusion Compared with 2003,HCV positive rate decreased significantly in 2020.HCV infected patients were mainly from the department of infectious diseases,followed by departments of gastroenterology,nephrology and general ICU.HCV infection positive rate varies with gender,age,and region.
		                        		
		                        		
		                        		
		                        	
3.Exogenous leptin improves cerebral ischemia-reperfusion-induced glutamate excitotoxic injury in mice by up-regulating GLT-1 and GLAST expression in astrocytes
Jie CHEN ; Chenxu LIU ; Chun WANG ; Li LI ; Weiting TAO ; Jingru XUN ; Honghui TANG ; Li HUANG
Journal of Southern Medical University 2024;44(6):1079-1087
		                        		
		                        			
		                        			Objective To investigate the protective effect of exogenous leptin against focal cerebral ischemia-reperfusion(I/R)injury in mice and explore the underlying mechanism.Methods A total of 100 C57BL/6 mice were randomly divided into 5 groups,including a sham-operated group,cerebral I/R model group,and 3 leptin treatment groups with intraperitoneal injections of 0.5,1.0 or 2.0 leptin immediately after occlusion of the internal carotid artery.At 24 h after reperfusion,neurological function scores of the mice were assessed,and TTC staining was used to determine the area of cerebral infarction.The pathological changes in the cortical brain tissue of the mice were observed using HE staining,and degenerative damage of the cortical neurons were assessed with Fluoro-Jade C staining.The expression of glial fibrillary acidic protein in cortical brain tissues was detected using immunohistochemistry and Western blotting.In another 45 C57BL/6 mice with sham operation,I/R modeling,or leptin(1 mg/kg)treatment,glutamic acid in the cortical brain tissue was detected using glutamate assay,and cortical glutamate-aspartate transporter(GLAST)and glutamate transporter-1(GLT-1)protein expressions were detected using immunohistochemistry.Results Compared with the I/R model mice,the leptin-treated mice had significantly lower neurological deficit scores,smaller cerebral infarct area,milder pathologies in the cortical brain tissue,and lessened cortical neuronal damage with normal morphology and less excessive proliferation of the astrocytes.Leptin treatment significantly up-regulated the expressions of GLT-1 and GLAST and lowered the content of glutamic acid in the brain tissue of the I/R mice.Conclusion Exogenous leptin has obvious neuroprotective effect against cerebral I/R injury in mice,mediated probably by controlling excessive astrocyte proliferation and up-regulating cortical GLT-1 and GLAST expressions to reduce glutamate-mediated excitotoxic injury of the astrocytes.
		                        		
		                        		
		                        		
		                        	
4.Effect of Huoxue Yiqi Decoction Combined with Pinggan Qianyang Muzu Powder on Blood Lipid Metabolism and Cardiac Function in Female Patients with Essential Hypertension
Xi-Xun LIAO ; Kai-Ge ZHOU ; Yue-Hua LI ; Jie CHEN
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(5):1123-1129
		                        		
		                        			
		                        			Objective To investigate the effects of Huoxue Yiqi Decoction(composed of Carthami Flos,Crataegi Fructus,Salviae Miltiorrhizae Radix et Rhizoma,and Astragali Radix)combined with Pinggan Qianyang Muzu Powder(composed of Uncariae Ramulus cum Uncis,Prunellae Spica,Chrysanthemi Flos,Lonicerae Japonicae Flos and Cassiae Semen)on blood lipid metabolism and cardiac function in female patients with essential hypertension.Methods A total of 220 female patients with essential hypertension of qi deficiency and blood stasis type were randomly divided into observation group and control group,with 110 cases in each group.The control group was treated with conventional western medicine,and the observation group was treated with Huoxue Yiqi Decoction orally combined with Pinggan Qianyang Muzu Powder for foot bath on the basis of treatment for the control group.The course of treatment for the two groups covered one month.Before and after treatment,the two groups were observed in the changes of blood pressure,blood lipid,traditional Chinese medicine(TCM)syndrome scores and cardiac function indicators of left ventricular ejection fraction(LVEF)and stroke volume(SV).After treatment,the clinical efficacy of the two groups was evaluated.Results(1)After one month of treatment,the total effective rate of the observation group was 95.45%(105/110),and that of the control group was 84.55%(93/110).The intergroup comparison(tested by chi-square test)showed that the efficacy of the observation group was superior to that of the control group(P<0.01).(2)After treatment,the TCM syndrome scores of primary symptoms and secondary symptoms in the two groups were decreased when compared with those before treatment(P<0.05),and the decrease in the observation group was superior to that in the control group(P<0.01).(3)After treatment,the levels of systolic blood pressure(SBP)and diastolic blood pressure(DBP)in the two groups were lower than those before treatment(P<0.05),and the decrease of SBP and DBP levels in the observation group was significantly superior to that in the control group(P<0.01).(4)After treatment,the levels of blood lipid indicators of total cholesterol(TC),triglyceride(TG)and low density lipoprotein cholesterol(LDL-C)in the two groups were lower than those before treatment(P<0.05),and the levels of high density lipoprotein cholesterol(HDL-C)were higher than those before treatment(P<0.05).The decrease of TC,TG and LDL-C levels and the increase of HDL-C level in the observation group were significantly superior to those in the control group(P<0.01).(5)After treatment,the levels of cardiac function indicators of LVEF and SV in the two groups were higher than those before treatment(P<0.05),and the increase of LVEF and SV levels in the observation group was significantly superior to that in the control group(P<0.01).Conclusion The combination of Huoxue Yiqi Decoction and Pinggan Qianyang Muzu Powder exerts certain effect in the treatment of female patients with essential hypertension of qi deficiency and blood stasis type.The combined therapy is effective on controlling blood pressure,improving blood lipid metabolism and cardiac function indicators.
		                        		
		                        		
		                        		
		                        	
5.Clinical Experience of JI Xun-Chao for the Syndrome Differentiation and Treatment of Cough Variant Asthma in Children Based on the Corresponding Predominant Two-Hour Periods of Meridians and Zang-Fu Viscera
Jie ZHENG ; Ji-Ling LIU ; Jing ZHANG ; Xun-Chao JI
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(11):3045-3049
		                        		
		                        			
		                        			This article introduces the clinical experience of Professor JI Xun-Chao for the syndrome differentiation and treatment of cough variant asthma(CVA)in children based on the corresponding predominant two-hour periods of meridians and zang-fu viscera.The corresponding predominant two-hour periods of meridians and zang-fu viscera are derived from the Ziwu Liuzhu theory(midnight-midday ebb and flow,the movement of qi and blood in the meridians following heavenly-stems and earthly-branches cycle),which reflects the relationship of the two-hour periods with qi and blood circulation in the meridians.Professor JI Xun-Chao believes that the attack of CVA in children is related with the two-hour periods,and has close relationship with the zang-fu viscera of lung,liver and spleen.Based on the clinical manifestation characteristics,children CVA can be divided into two types,namely"jiming cough"(cough attack or aggravation at 23∶00-3∶00)and"pingdan cough"(cough attack or aggravation at 3∶00-11∶00),and jiming cough can develop into pingdan cough with the transformation of the pathogenesis.For the treatment of jiming cough,therapies of soothing liver to check upward adverse flow of qi and warming lung to resolve phlegm were usually adopted,and Wenfei Jiangni Decoction was prescribed.For the treatment of pingdan cough,therapies of invigorating spleen and replenishing qi together with the clearance of residual pathogens were adopted,and Tufuling Decoction was prescribed.This article summarized the temporal-differentiation experience of Professor JI Xun-Chao in treating children CVA based on the corresponding predominant two-hour periods of meridians and zang-fu viscera and the therapeutic principle of treating symptom and root cause simultaneously,which will provide reference for clinical treatment of children CVA.
		                        		
		                        		
		                        		
		                        	
6.Moxifloxacin treatment for Mycoplasma hominis meningitis in an extremely preterm infant
Wei-Ying MAO ; Jiang-Er LAN ; Ming-Yu GAN ; Xun-Jie ZHANG ; Hui YU ; Li-Yuan HU ; Rong ZHANG ; Yun CAO ; Mi-Li XIAO
Chinese Journal of Contemporary Pediatrics 2024;26(4):432-436
		                        		
		                        			
		                        			The patient,a male newborn,was admitted to the hospital 2 hours after birth due to prematurity(gestational age 27+5 weeks)and respiratory distress occurring 2 hours postnatally.After admission,the infant developed fever and elevated C-reactive protein levels.On the fourth day after birth,metagenomic next-generation sequencing of cerebrospinal fluid indicated a positive result for Mycoplasma hominis(9 898 reads).On the eighth day,a retest of cerebrospinal fluid metagenomics confirmed Mycoplasma hominis(56 806 reads).The diagnosis of purulent meningitis caused by Mycoplasma hominis was established,and the antibiotic treatment was switched to moxifloxacin[5 mg/(kg·day)]administered intravenously for a total of 4 weeks.After treatment,the patient's cerebrospinal fluid tests returned to normal,and he was discharged as cured on the 76th day after birth.This article focuses on the diagnosis and treatment of neonatal Mycoplasma hominis purulent meningitis,introducing the multidisciplinary diagnosis and treatment of the condition in extremely preterm infants.[Chinese Journal of Contemporary Pediatrics,2024,26(4):432-436]
		                        		
		                        		
		                        		
		                        	
7.Antimicrobial resistance of bacteria from blood specimens:surveillance re-port from Hunan Province Antimicrobial Resistance Surveillance System,2012-2021
Hong-Xia YUAN ; Jing JIANG ; Li-Hua CHEN ; Chen-Chao FU ; Chen LI ; Yan-Ming LI ; Xing-Wang NING ; Jun LIU ; Guo-Min SHI ; Man-Juan TANG ; Jing-Min WU ; Huai-De YANG ; Ming ZHENG ; Jie-Ying ZHOU ; Nan REN ; An-Hua WU ; Xun HUANG
Chinese Journal of Infection Control 2024;23(8):921-931
		                        		
		                        			
		                        			Objective To understand the change in distribution and antimicrobial resistance of bacteria isolated from blood specimens of Hunan Province,and provide for the initial diagnosis and treatment of clinical bloodstream infection(BSI).Methods Data reported from member units of Hunan Province Antimicrobial Resistance Survei-llance System from 2012 to 2021 were collected.Bacterial antimicrobial resistance surveillance method was imple-mented according to the technical scheme of China Antimicrobial Resistance Surveillance System(CARSS).Bacteria from blood specimens and bacterial antimicrobial susceptibility testing results were analyzed by WHONET 5.6 soft-ware and SPSS 27.0 software.Results A total of 207 054 bacterial strains were isolated from blood specimens from member units in Hunan Province Antimicrobial Resistance Surveillance System from 2012 to 2021,including 107 135(51.7%)Gram-positive bacteria and 99 919(48.3%)Gram-negative bacteria.There was no change in the top 6 pathogenic bacteria from 2012 to 2021,with Escherichia coli(n=51 537,24.9%)ranking first,followed by Staphylococcus epidermidis(n=29 115,14.1%),Staphylococcus aureus(n=17 402,8.4%),Klebsiella pneu-moniae(17 325,8.4%),Pseudomonas aeruginosa(n=4 010,1.9%)and Acinetobacter baumannii(n=3 598,1.7%).The detection rate of methicillin-resistant Staphylococcus aureus(MRSA)decreased from 30.3%in 2015 to 20.7%in 2021,while the detection rate of methicillin-resistant coagulase-negative Staphylococcus(MRCNS)showed an upward trend year by year(57.9%-66.8%).No Staphylococcus was found to be resistant to vancomy-cin,linezolid,and teicoplanin.Among Gram-negative bacteria,constituent ratios of Escherichia coli and Klebsiella pneumoniae were 43.9%-53.9%and 14.2%-19.5%,respectively,both showing an upward trend(both P<0.001).Constituent ratios of Pseudomonas aeruginosa and Acinetobacter baumannii were 3.6%-5.1%and 3.0%-4.5%,respectively,both showing a downward trend year by year(both P<0.001).From 2012 to 2021,resistance rates of Escherichia coli to imipenem and ertapenem were 1.0%-2.0%and 0.6%-1.1%,respectively;presenting a downward trend(P<0.001).The resistant rates of Klebsiella pneumoniae to meropenem and ertapenem were 7.4%-13.7%and 4.8%-6.4%,respectively,presenting a downward trend(both P<0.001).The resistance rates of Pseudomonas aeruginosa and Acinetobacter baumannii to carbapenem antibiotics were 7.1%-15.6%and 34.7%-45.7%,respectively.The trend of resistance to carbapenem antibiotics was relatively stable,but has de-creased compared with 2012-2016.The resistance rates of Escherichia coli to the third-generation cephalosporins from 2012 to 2021 were 41.0%-65.4%,showing a downward trend year by year.Conclusion The constituent ra-tio of Gram-negative bacillus from blood specimens in Hunan Province has been increasing year by year,while the detection rate of carbapenem-resistant Gram-negative bacillus remained relatively stable in the past 5 years,and the detection rate of coagulase-negative Staphylococcus has shown a downward trend.
		                        		
		                        		
		                        		
		                        	
8.Properties of gelatin-polyethylene glycol hydrogel loaded with silver nanoparticle Chlorella and its effects on healing of infected full-thickness skin defect wounds in mice
Zhenzhen YAN ; Yuxiang WANG ; Tinglin ZHANG ; Jingnan XUN ; Yicheng MA ; Chao JI ; Jie GAO ; Shichu XIAO
Chinese Journal of Burns 2024;40(1):33-42
		                        		
		                        			
		                        			Objective:To explore the properties of gelatin-polyethylene glycol hydrogel loaded with silver nanoparticle (AgNP) Chlorella (hereinafter referred to as the composite hydrogel) and its effects on healing of infected full-thickness skin defect wounds in mice. Methods:The research was an experimental research. The simple gelatin-polyethylene glycol hydrogel (hereinafter referred to as the simple hydrogel) and the composite hydrogel were prepared, and the appearance and injectability of the two hydrogels were observed at 55 and 37 ℃, and under the irradiation of 808 nm near-infrared light, respectively. An electronic universal testing machine was employed to assess the tensile and compressive stress-strain properties of both types of hydrogels at room temperature. Additionally, the cyclic compressive stress-strain properties of the composite hydrogel were examined at 80% of the maximum compressive stress. Staphylococcus aureus or Escherichia coli solution was added to phosphate buffer solution (PBS), simple hydrogel, and composite hydrogel, respectively. The part of composite hydrogel containing Staphylococcus aureus or Escherichia coli solution was irradiated with near-infrared light for 5 minutes. After each sample was incubated for 6 h, the dilution plating method was used to detect and calculate the mortality rates of the two bacteria at 24 h of culture ( n=5). The discarded foreskin tissue was taken from a 6-year-old healthy boy admitted to the Department of Urology of the First Affiliated Hospital of Naval Medical University for circumcision. Primary human fibroblasts (HFbs) were isolated using the enzyme extraction method, routinely cultured to the 3 rd to 6 th passages for subsequent cellular experiments. Composite hydrogel extracts with final mass concentrations of 100.0, 50.0, 25.0, 12.5, and 0 mg/mL were respectively prepared and used to culture HFbs, and the cell proliferation after 24 h of culture was detected using a cell counting kit 8 ( n=3). A total of twenty 6-8 weeks old C57BL/6J female mice were utilized, and a full-thickness skin defect was surgically created on the back of each mouse. The wounds were infected with Staphylococcus aureus solution. The infected mice were divided into blank control group, simple hydrogel group, composite hydrogel group, and combined treatment group according to the random number table, and the wounds were treated with PBS, simple hydrogel, composite hydrogel, and composite hydrogel+light irradiation (under the irradiation of 808 nm near-infrared light for 5 min), respectively, with 5 mice in each group. On post injury day (PID) 0 (immediately after the first wound treatment), 3, 7, and 14, an overall assessment of wound exudation and healing were conducted, and the wound healing rates on PID 7 and 14 were calculated ( n=5). On PID 14, hematoxylin-eosin staining was performed to observe histopathological changes in the mouse wound. Results:Both simple hydrogel and composite hydrogel were in a solution state at 55 ℃ and transition to a gel state when cooling to 37 ℃. After the two hydrogels were irradiated by near-infrared light, only the composite hydrogel reheated up and returned to the solution state again with injectability. The maximum tensile stress of the composite hydrogel was up to 301.42 kPa, with a corresponding strain of 87.19%; the maximum compressive stress was up to 413.79 kPa, with a corresponding strain of 91.67%, which was similar to the tensile and compressive properties of the simple hydrogel. After 10 compression cycles, the maximum compressive stress of the composite hydrogel still reached 84.1% of the first compressive stress. After 24 h of culture, the mortality rate of Staphylococcus aureus treated with simple hydrogel was significantly higher than that treated with PBS ( P<0.05); the mortality rates of Escherichia coli and Staphylococcus aureus treated with composite hydrogel alone were significantly higher than those treated with simple hydrogel ( P<0.05); the mortality rates of Escherichia coli and Staphylococcus aureus treated with composite hydrogel+light irradiation were significantly higher than those treated with composite hydrogel alone ( P<0.05). After 24 h of culture, compared with that cultured in composite hydrogel immersion solution with final mass concentration of 0 mg/mL, the proliferation activity of HFbs cultured in composite hydrogel immersion solution with final mass concentrations of 25.0 and 50.0 mg/mL was significantly enhanced ( P<0.05), while the proliferation activity of HFbs cultured in composite hydrogel immersion solution with final mass concentration of 100 mg/mL was significantly decreased ( P<0.05). On PID 0 and 3, more purulent secretions were seen in the wounds of mice in blank control group and simple hydrogel group, while only a small amount of exudate was observed in the wounds of mice in composite hydrogel group, and no obvious infection was observed in the wounds of mice in combined treatment group. On PID 7 and 14, the wound healing rates of mice in simple hydrogel group were significantly higher than those in blank control group ( P<0.05); the wound healing rates of mice in composite hydrogel group were significantly higher than those in simple hydrogel group ( P<0.05); the wound healing rates in combined treatment group were significantly higher than those in composite hydrogel group ( P<0.05). On PID 14, the wounds of mice in blank control group exhibited a high infiltration of inflammatory cells with no new epithelial layer observed; the wounds of mice in simple hydrogel group displayed a short length of newly formed epithelium with a small amount of inflammatory cells; the wounds of mice in composite hydrogel group exhibited continuous formation of new epithelium and a large amount of immature granulation tissue; the wounds of mice in combined treatment group showed continuous epithelialization with less immature granulation tissue. Conclusions:The prepared composite hydrogel exhibits excellent thermosensitivity, photothermal properties, and injectability, as well as excellent mechanical properties, antibacterial properties, and biocompatibility, and can promote the healing of infected full-thickness skin defect wounds in mice.
		                        		
		                        		
		                        		
		                        	
9.Exogenous leptin improves cerebral ischemia-reperfusion-induced glutamate excitotoxic injury in mice by up-regulating GLT-1 and GLAST expression in astrocytes
Jie CHEN ; Chenxu LIU ; Chun WANG ; Li LI ; Weiting TAO ; Jingru XUN ; Honghui TANG ; Li HUANG
Journal of Southern Medical University 2024;44(6):1079-1087
		                        		
		                        			
		                        			Objective To investigate the protective effect of exogenous leptin against focal cerebral ischemia-reperfusion(I/R)injury in mice and explore the underlying mechanism.Methods A total of 100 C57BL/6 mice were randomly divided into 5 groups,including a sham-operated group,cerebral I/R model group,and 3 leptin treatment groups with intraperitoneal injections of 0.5,1.0 or 2.0 leptin immediately after occlusion of the internal carotid artery.At 24 h after reperfusion,neurological function scores of the mice were assessed,and TTC staining was used to determine the area of cerebral infarction.The pathological changes in the cortical brain tissue of the mice were observed using HE staining,and degenerative damage of the cortical neurons were assessed with Fluoro-Jade C staining.The expression of glial fibrillary acidic protein in cortical brain tissues was detected using immunohistochemistry and Western blotting.In another 45 C57BL/6 mice with sham operation,I/R modeling,or leptin(1 mg/kg)treatment,glutamic acid in the cortical brain tissue was detected using glutamate assay,and cortical glutamate-aspartate transporter(GLAST)and glutamate transporter-1(GLT-1)protein expressions were detected using immunohistochemistry.Results Compared with the I/R model mice,the leptin-treated mice had significantly lower neurological deficit scores,smaller cerebral infarct area,milder pathologies in the cortical brain tissue,and lessened cortical neuronal damage with normal morphology and less excessive proliferation of the astrocytes.Leptin treatment significantly up-regulated the expressions of GLT-1 and GLAST and lowered the content of glutamic acid in the brain tissue of the I/R mice.Conclusion Exogenous leptin has obvious neuroprotective effect against cerebral I/R injury in mice,mediated probably by controlling excessive astrocyte proliferation and up-regulating cortical GLT-1 and GLAST expressions to reduce glutamate-mediated excitotoxic injury of the astrocytes.
		                        		
		                        		
		                        		
		                        	
10.Mechanism of Dendrobium huoshanense Polysaccharide Against Neuroinflammation in Parkinson's Disease Model: Based on NLRP3 Inflammasome
Xiaolong SHANG ; Chenchen XU ; Jianjian DONG ; Chunling CI ; Pei ZHANG ; Liangjie ZHANG ; Hongyang JIN ; Jie LI ; Xun WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(11):97-105
		                        		
		                        			
		                        			ObjectiveTo explore the mechanism of Dendrobium huoshanense polysaccharide (DHP) against inflammatory damage of neurons in Parkinson's disease (PD) model. MethodSH-SY5Y cells were randomized into blank group, model group, and DHP group. The survival rate of cells was measured by thiazole blue(MTT) assay, and the levels of lactate dehydrogenase (LDH), reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were measured by colorimetric analysis. BV-2 microglia were classified into blank group, model group, DHP group, and MCC950 group (positive control group), and enzyme-linked immunosorbent assay (ELISA) was applied to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-18 (IL-18). The expression of NOD-like receptor protein 3 (NLRP3), adaptor protein apoptosis-associated dot protein (ASC), cysteine aspartic protease-1 (Caspase-1), and IL-1β was measured by Western blot. A total of 50 C57BL/6 mice were randomized into blank group, model group, DHP low-dose (100 mg·kg-1) group, DHP equivalent-dose (350 mg·kg-1) group, and MCC950 group (positive control group), 10 mice in each group. The motor balance and coordination of C57BL/6 mice were observed by beam walking test, tail suspension test and rotarod test. The levels of Iba-1 and tyrosine hydroxylase (TH) were detected by immunofluorescence staining. The damage of dopaminergic neurons in the substantia nigra was detected by FJB staining. The levels of inflammatory factors such as IL-1β, IL-18, and TNF-α in mouse midbrain tissues were detected by ELISA and the protein levels of NLRP3, ASC, Caspase-1, and IL-1β protein were measured by Western blot. ResultCompared with the blank group, the SH-SY5Y model group showed decreased cell survival, increased levels of LDH, ROS, and MDA (P<0.05), and decreased levels of SOD (P<0.05). Compared with the model group, the DHP group demonstrated increased cell survival, decreased levels of LDH, ROS, and MDA (P<0.01), and increased level of SOD (P<0.01). Compared with the blank group, BV-2 model group had high levels of IL-1β, IL-18, and TNF-α (P<0.05) and high protein expression of NLRP3, Caspase-1, IL-1β, and ASC (P<0.05). Compared with the model group, DHP and MCC950 groups demonstrated low levels of IL-1β, IL-18, and TNF-α (P<0.01) and low protein expression of NLRP3, Caspase-1, IL-1β, and ASC (P<0.01). Compared with the blank group, the C57BL/6 model group displayed long time to pass the balance wood (P<0.05), short time spent on the rod in the rotarod test (P<0.05), high levels of IL-1β, IL-18, and TNF-α (P<0.05) and expression of Iba-1 in the midbrain substantia nigra (P<0.05), low TH expression (P<0.05), more positive neurons in the FJB staining (P<0.05), and high expression of NLRP3, Caspase-1, ASC, and IL-1β proteins (P < 0.05). Compared with the model group, the mice in the DHP and MCC950 groups had short time to pass the balance beam (P<0.01), long time spent on the rod (P<0.01), low levels of IL-1β, IL-18, and TNF-α (P<0.01), low Iba-1 expression in midbrain substantia nigra (P<0.01), high TH expression (P<0.01), and small number of positive neurons in the midbrain substantia nigra (P<0.01). The expression of NLRP3, ASC, and IL-1β proteins was lower in the MCC950 group (P<0.01), and the expression of NLRP3, ASC, Caspase-1 and IL-1β proteins was lower in the DHP equivalent-dose group (P<0.01) than in the model group. ConclusionDHP has anti-oxidative stress effect. It regulates the expression of NLRP3 inflammasome and inhibits the overactivation of microglia, thereby alleviating the neuroinflammatory injury in PD and exerting the neuroprotective effect. 
		                        		
		                        		
		                        		
		                        	
            
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