1.Identification of Dalbergia odorifera and Its Counterfeits by HS-GC-MS
Li ZHAO ; Xiaowei MENG ; Jiarong LI ; Qing ZHU ; Xianwen WEI ; Ronghua LIU ; Lanying CHEN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(2):156-163
ObjectiveTo screen the differential markers by analyzing volatile components in Dalbergia odorifera and its counterfeits, in order to provide reference for authentication of D. odorifera. MethodThe volatile components in D. odorifera and its counterfeits were detected by headspace gas chromatography-mass spectrometry(HS-GC-MS), and the GC conditions were heated by procedure(the initial temperature of the column was 50 ℃, the retention time was 1 min, and then the temperature was raised to 300 ℃ at 10 ℃ for 10 min), the carrier gas was helium, and the flow rate was 1.0 mL·min-1, the split ratio was 10∶1, and the injection volume was 1 mL. The MS conditions used electron bombardment ionization(EI) with the scanning range of m/z 35-550. The compound species were identified by database matching, the relative content of each component was calculated by the peak area normalization method, and principal component analysis(PCA), orthogonal partial least squares-discrimination analysis(OPLS-DA) and cluster analysis were performed on the detection results by SIMCA 14.1 software, and the differential components of D. odorifera and its counterfeits were screened out according to the variable importance in the projection(VIP) value>2 and P<0.05. ResultA total of 26, 17, 8, 22, 24 and 7 volatile components were identified from D. odorifera, D. bariensis, D. latifolia, D. benthamii, D. pinnata and D. cochinchinensis, respectively. Among them, there were 11 unique volatile components of D. odorifera, 6 unique volatile components of D. bariensis, 3 unique volatile components of D. latifolia, 6 unique volatile components of D. benthamii, 8 unique volatile components of D. pinnata, 4 unique volatile components of D. cochinchinensis. The PCA results showed that, except for D. latifolia and D. cochinchinensis, which could not be clearly distinguished, D. odorifera and other counterfeits could be distributed in a certain area, respectively. The OPLS-DA results showed that D. odorifera and its five counterfeits were clustered into one group each, indicating significant differences in volatile components between D. odorifera and its counterfeits. Finally, a total of 31 differential markers of volatile components between D. odoriferae and its counterfeits were screened. ConclusionHS-GC-MS combined with SIMCA 14.1 software can systematically elucidate the volatile differential components between D. odorifera and its counterfeits, which is suitable for rapid identification of them.
2.Improved discharge survival in pre-hospital cardiac arrest patients: the Shenzhen Bao'an experience
Wenwu ZHANG ; Jinfeng LIANG ; Qingli DOU ; Jun XU ; Jinle LIN ; Conghua WANG ; Wuyuan TAO ; Xianwen HUANG ; Wenhua LIU ; Yujie LI ; Xiaoming ZHANG ; Cuimei XING ; Huadong ZHU ; Xuezhong YU
Chinese Journal of Emergency Medicine 2024;33(11):1518-1523
Objective:Cardiac arrest (CA) represents a significant public health challenge, posing a substantial threat to individual health and survival. To enhance the survival rates of patients experiencing out-of-hospital cardiac arrest (OHCA), Baoan District in Shenzhen City has undertaken exploratory initiatives and practical interventions, yielding promising preliminary outcomes.Methods:1.Innovate emergency medical services by developing a "four-circle integration" system that connects to the hospital. This system encompasses the social emergency medical system, the out-of-hospital emergency medical system, the in-hospital emergency medical service system, and the intensive care treatment system. 2.Develop a comprehensive model for the construction of a social emergency medical training system, characterized by party leadership, government oversight, departmental coordination, professional guidance, technological support, and community involvement, termed the "Baonan Model." Additionally, establish evaluation criteria to assess the effectiveness of the social emergency medical training system in Baonan District; 3. Develop a cardiac arrest registration system and a social emergency medical training management system for Baonan District; 4. Enhance the proficiency in treatment techniques and the quality of cardiopulmonary resuscitation among emergency medical professionals; 5. Strengthen and advance the development of a "five-minute social rescue network" to address the critical "emergency window period." .Result:In Baonan District, 9.18% of the public is trained in emergency medical skills. The bystander CPR rate for OHCA is 26.11%, AED use is at 4.78%, the 30-day survival rate is 6.31%, and the discharge survival rate is 4.44%.Conclusion:The implementation of the aforementioned measures can substantially enhance the survival rate of patients experiencing OHCA at the time of discharge.
3.The interaction between polyphyllin I and SQLE protein induces hepatotoxicity through SREBP-2/HMGCR/SQLE/LSS pathway
Zhiqi LI ; Qiqi FAN ; Meilin CHEN ; Ying DONG ; Farong LI ; Mingshuang WANG ; Yulin GU ; Simin GUO ; Xianwen YE ; Jiarui WU ; Shengyun DAI ; Ruichao LIN ; Chongjun ZHAO
Journal of Pharmaceutical Analysis 2023;13(1):39-54
Polyphyllin Ⅰ(PPⅠ)and polyphyllin Ⅱ(PⅡ)are the main active substances in the Paris polyphylla.However,liver toxicity of these compounds has impeded their clinical application and the potential hepatotoxicity mechanisms remain to be elucidated.In this work,we found that PPⅠ and PⅡ exposure could induce significant hepatotoxicity in human liver cell line L-02 and zebrafish in a dose-dependent manner.The results of the proteomic analysis in L-02 cells and transcriptome in zebrafish indicated that the hepa-totoxicity of PPⅡ and PⅡwas associated with the cholesterol biosynthetic pathway disorders,which were alleviated by the cholesterol biosynthesis inhibitor lovastatin.Additionally,3-hydroxy-3-methy-lglutaryl CoA reductase(HMGCR)and squalene epoxidase(SQLE),the two rate-limiting enzymes in the choles-terol synthesis,selected as the potential targets,were confirmed by the molecular docking,the over-expression,and knockdown of HMGCR or SQLE with siRNA.Finally,the pull-down and surface plasmon resonance technology revealed that PPⅠ could directly bind with SQLE but not with HMGCR.Collectively,these data demonstrated that PPⅠ-induced hepatotoxicity resulted from the direct binding with SQLE protein and impaired the sterol-regulatory element binding protein 2/HMGCR/SQLE/lanosterol synthase pathways,thus disturbing the cholesterol biosynthesis pathway.The findings of this research can contribute to a better understanding of the key role of SQLE as a potential target in drug-induced hepatotoxicity and provide a therapeutic strategy for the prevention of drug toxic effects with similar structures in the future.
4.Correlation between voluntary blood donation and autonomous motivation
Journal of Preventive Medicine 2023;35(9):786-789
Objective:
To explore the relationship between voluntary blood donation and autonomous motivation based on self-determination theory, so as to provide insights into optimizing voluntary blood donation recruitment.
Methods:
Participants meeting the blood donation criteria were selected from blood stations and medical institutions in Hangzhou City, Jiaxing City and Quzhou City, Zhejiang Province. A self-designed questionnaire based on self-determination theory was used to collect basic information, awareness and source of blood donation knowledge, autonomous motivation and controlled motivation. The correction between autonomous motivation and voluntary blood donation was analyzed using a multivariable logistic regression model.
Results:
A total of 986 participants at ages of 18-55 years were surveyed, including 730 females (74.04%) and 256 males (25.96%). Among them, 705 participants (71.50%) had experience of voluntary blood donation. The mean score for blood donation knowledge was (5.76±1.17) points, and the total motivation score for blood donation was (58.21±8.17) points, with autonomous motivation scoring (40.86±5.46) points and controlled motivation scoring (17.35±3.26) points. Multivariable logistic regression analysis identified gender (female, OR=0.419, 95%CI: 0.273-0.644), occupation (medical profession, OR=4.027, 95%CI: 2.440-6.646), blood donation knowledge (OR=1.307, 95%CI: 1.159-1.475) and autonomous motivation (OR=1.074, 95%CI: 1.033-1.116) as factors affecting voluntary blood donation.
Conclusions
Autonomous motivation may promote voluntary blood donation. In blood donation recruitment efforts, it is vital to enhance autonomous motivation and make appropriate use of controlled motivation.
5.Assessment of conventional magnetic resonance morphological measurements in diagnosis and differential diagnosis of progressive supranuclear paralysis
Tieyu WU ; Chuan LI ; Yue JIAO ; Juan LI ; Tong WU ; Shangpei WANG ; Xianwen CHEN
Chinese Journal of Geriatrics 2023;42(2):188-195
Objective:To study the features as well as the diagnosis and differential diagnosis values by conventional MRI morphometrics in different clinical subtypes of progressive supranuclear palsy(PSP).Methods:Forty five patients with PSP were included, comprising three PSP subtypes: 15 cases of Richardson's syndrome(PSP-RS), 15 cases of Parkinson's syndrome(PSP-P)and 15 cases of progressive frozen gait(PSP-PFG). In addition, three control groups were established: 15 cases of multiple system atrophy-Parkinson's syndrome(MSA-P), 30 cases of primary Parkinson's disease(PD)and 40 healthy controls(HC). Midbrain area-to-Pons area ratio(M/P), Magnetic Resonance Parkinsonism Index(MRPI, MRPI2.0), width ratio of middle cerebellar peduncle to superior cerebellar peduncle(MCP/SCP), Midbrain-to-Pons ratio(MTPR), Angle of cerebral peduncle(A cp), third ventricle width/frontal horns width ratio(V 3rd/FH), and Humming bird sign rating scale(HBS-RS)scores were calculated.Diagnostic sensitivity and specificity were performed by ROC curve to assess the accuracy of these imaging indicators in the diagnosis and differential diagnosis of PSP and its subtypes. Results:The MRPI, MRPI2.0, MCP/SCP and HBS-RS scores were significantly higher in PSP group than in other control groups( H=69.351, 66.776, 33.926 and 84.694, all P<0.05), while M/P and MTPR were significantly lower in PSP group than in other control groups(H=60.101 and 77.276, all P<0.05). PSP group also had higher V 3rd/FH compared with PD or HC group( F=17.168, P<0.05), but not with MSA-P group( Z=-1.602, P>0.05). The above differences also existed between each PSP subgroup and control groups.Among PSP subgroups, PSP-PFG subgroup had a larger A cp than did PSP-RS( Z=-2.510, P<0.05), and had higher HBS-RS score than did PSP-P group( Z=-2.380, P<0.05). No significant differences in other MRI morphometric indexes were identified among PSP subtypes.The M/P, MRPI, MTPR, MRPI2.0, HBS-RS score showed good accuracy in diagnosing PSP and its each subgroup, with HBS-RS score being the most accurate indicator, when the cutoff value was 2, the AUC values were all higher than 0.99, and the sensitivity and specificity were all above 90%.PSP and its subtypes were best distinguished from MSA-P by MRPI, when the cutoff value was 9.94, the AUC values were all higher than 0.90, with the sensitivity of 100% and specificity of 86.67%.PSP and its subtypes were best distinguished from PD by MTPR, AUC values were all above 0.95, with slightly different cutoff values.Almost all the morphological measurement parameters failed to show significant sensitivity and specificity in discriminating subtypes of PSP.The sensitivity and specificity of almost all MRI morphometry indicators in differentiating different subtypes of PSP are not high. Conclusions:MRI morphometrics have a high value both in the diagnosis of PSP and its subtypes, and also in specific application fields.MRI morphometrics have a limited value in discriminating PSP subtypes.
6.Role of SIRT1/NLRP3 signaling pathway in sevoflurane postconditioning-induced attenuation of oxygen-glucose deprivation and restoration injury in HT22 cells
Xiaojing WAN ; Li ZHANG ; Su HU ; Yujie WU ; Zhilun NIU ; Yiming ZHANG ; Xianwen HU
Chinese Journal of Anesthesiology 2023;43(6):741-745
Objective:To evaluate the role of silent information regulator-1 (SIRT1)/nucleotide-binding domain (NOD)-like receptor protein-3 (NLRP3) signaling pathway in sevoflurane postconditioning-induced attenuation of oxygen-glucose deprivation and restoration (OGD/R) injury in mouse hippocampal neuronal cell line (HT22) cells.Methods:The HT22 cells were seeded in a culture plate (96-well plate, 100 μl/well; 6-well plate, 2 ml/well) at the density of 5×10 4 cells/ml or in a culture dish (6 cm in diameter) and then divided into 4 groups ( n=24 each) using a random number table method: control group (Control group), OGD/R group, sevoflurane postconditioning group (SPC group), and SIRT1 small interfering RNA group (si-SIRT 1 group). In Control group, cells were cultured at 37 ℃ in normal culture atmosphere. In OGD/R group, the culture medium was replaced with glucose-free serum-free culture medium, and cells were exposed to 95% N 2+ 5% CO 2 for 4 h in an incubator at 37 ℃, and then the glucose-free serum-free culture medium was replaced with the primary culture medium, and cells were cultured for 24 h at 37 ℃ in normal culture atmosphere. In SPC group, the glucose-free serum-free culture medium was replaced with the primary cell culture medium after 4-h oxygen and glucose deprivation, the cells were put into the hypoxia incubator chamber which was filled with 2% sevoflurane immediately after start of reoxygenation, then the chamber was placed in an incubator and the cells were cultured for 1 h at 37 ℃ in normal culture atmosphere, and finally the cells were removed from the chamber and cultured for 23 h at 37 ℃ in normal culture atmosphere. In si-SIRT1 group, SIRT1 small interfering RNA 150 pmol was added at 24 h before surgery, cells were then incubated, and the other procedures were the same as those previously described in group SPC. The cell survival rate was determined using MTT assay. TUNEL assay was used to detect cell apoptosis, and the apoptosis rate was calculated. The expression of SIRT1, NLRP3, IL-1β and IL-18 mRNA was determined using polymerase chain reaction. The expression of SIRT1, NLRP3, interleukin-1beta (IL-1β) and IL-18 was detected using Western blot. Results:Compared with Control group, the cell survival rate was significantly decreased, the apoptosis rate was increased, the expression of SIRT1 protein and mRNA was down-regulated, and the expression of NLRP3, IL-1β and IL-18 protein and mRNA was up-regulated in OGD/R group ( P<0.05). Compared with OGD/R group, the cell survival rate was significantly increased, the apoptosis rate was decreased, the expression of SIRT1 protein and mRNA was up-regulated, and the expression of NLRP3, IL-1β and IL-18 protein and mRNA was down-regulated in SPC group ( P<0.05). Compared with SPC group, the cell survival rate was significantly decreased, the apoptosis rate was increased, the expression of SIRT1 protein and mRNA was down-regulated, and the expression of NLRP3, IL-1β and IL-18 protein and mRNA was up-regulated in si-SIRT1 group ( P<0.05). Conclusions:Activation of SIRT1-NLRP3 signaling pathway is involved in sevoflurane postconditioning-induced attenuation of OGD/R injury in HT22 cells.
7.The role of GSK-3 β in Zebrafish cerebral hypoxia / reoxygenation injury and its effect on microtubule-associated protein 2
Mengsi Yang ; Li Zhang ; Xianwen Hu
Acta Universitatis Medicinalis Anhui 2023;58(2):202-208
Objective:
To investigate the effect of glycogen synthase kinase 3 β ( GSK-3 β) and its correlation with microtubule-associated protein 2 (MAP2) during cerebral hypoxia / reoxygenation (H / R) in zebrafish.
Methods:
The cerebral hypoxia / reoxygenation model of zebrafish was established.Healthy adult zebrafishes of the same size were divided into control group ( Control) ,hypoxia / reoxygenation group ( H / R) and hypoxia / reoxygenation + GSK-3 β inhibitor group (H / R + TDZD-8) for experiment.The brain tissues of zebrafish in each group were selected to determine the mRNA expressions of hypoxia inducible factor 1 αa and 1 αb (HiF-1αa and HIF-1 αb) at different reoxygenation time points by qRT-PCR , and the protein expression levels of HIF-1α , GSK-3 β , p-GSK-3 β (Ser 9) and MAP2 were detected by Western blot,TTC staining and TUNEL staining were used to detect cerebral infarction area and cell apoptosis ,and immunofluorescence was used to detect the distribution and expression of MAP2 in brain.
Results:
Compared with Control group,the mRNA levels of Hif-1αa and Hif-1 αb(P<0. 01) and protein expression of Hif-1 α(P<0. 01) increased in H / R group,the area of cerebral infarction (P <0. 01) and apoptotic cells(P <0. 01) increased,p-GSK-3 β ( Ser 9) / GSK-3 β ratio,MAP2 protein expression (P <0. 05) and immunofluorescence expression of MAP2 (P <0. 01 ) reduced ; Furthermore,TDZD-8 pretreatment could relieve the brain injury of H / R zebrafish by decreasing the infarct size and cell apoptosis,improving the ratio of p-GSK-3 β ( Ser 9 ) / GSK-3 β , and increasing the expression of MAP2.
Conclusion
Hypoxia / reoxygenation can cause brain neuron damage in zebrafish,and its mechanism may be related to inhibition of GSK-3 β phosphorylation and MAP2 expression.GSK-3 β specific inhibitor TDZD-8 can reverse the damage of brain neurons caused by hypoxia / reoxygenation by promoting the expression of P-GSK-3 β (Ser 9) and reducing MAP2 degradation.
8.Relationship between METTL3-mediated m6A methylation modification and SIRT1 during sevoflurane post-conditioning-induced mitigation of cognitive impairments in a mouse model of hemorrhagic shock and resuscitation
Yujie WU ; Li ZHANG ; Hui TAO ; Su HU ; Zhilun NIU ; Xiaojing WAN ; Xianwen HU
Chinese Journal of Anesthesiology 2023;43(11):1386-1391
Objective:To evaluate the relationship between methyltransferase-like 3(METTL3)-mediated RNA N6-Methyladenosine (m6A) methylation modification and silent information regulator factor 1 (SIRT1) during sevoflurane post-conditioning-induced mitigation of cognitive impairments in a mouse model of hemorrhagic shock and resuscitation(HSR).Methods:Forty clean-grade healthy male C57BL/6 mice, aged 8-10 weeks, with a body weight ranging from 22-26 g, were assigned into 5 groups ( n=8 each) using a random number table method: sham operation group, HSR group, sevoflurane post-conditioning + HSR group (SP+ HSR group), over-expression of METTL3 gene rAAV + sevoflurane post-conditioning + HSR group (METTL3+ SP+ HSR group), and over-expression of METTL3 gene rAAV negative control + sevoflurane post-conditioning + HSR group (NC+ SP+ HSR group). The HSR model was established by withdrawing 40% of the total blood volume from mice through the right carotid artery within 30 min, followed by reinfusion of the withdrawn blood over 30 min 1 h later. The SP+ HSR group underwent HSR modeling first and then inhaled sevoflurane (end-tidal concentration 2.4%) for 30 min starting from the time point immediately after blood transfusion. The Sham group and HSR group inhaled a mixture of 70% O 2 and 30% CO 2 for 30 min at the corresponding time points. In METTL3+ SP+ HSR group and NC+ SP+ HSR group, the corresponding virus 450 nl was injected into bilateral hippocampus at 4 weeks before establishing the model.Morris water maze and novel object recognition tests were conducted at 72 h after developing the model to assess the learning and memory abilities. After the end of behavioral tests, the expression of METTL3 and SIRT1 in hippocampal tissues was detected using Western blot, the expression of SIRT1 mRNA was measured using qRT-PCR, and the methylation of RNA m6A was detected using Dot blot. Results:Compared to Sham group, the escape latency was significantly prolonged at 1-6 days, the time spent in the target quadrant was shortened, the number of crossing the original platform was decreased, the novel object recognition index was decreased, the expression of METTL3 was up-regulated, the expression of SIRT1 protein and mRNA was down-regulated, and the methylation of RNA m6A was increased in HSR group( P<0.05). Compared to HSR group, the escape latency was significantly shortened at 1-6 days, the time spent in the target quadrant was prolonged, the number of crossing the original platform was increased, the novel object recognition index was increased, the expression of METTL3 was up-regulated, the expression of SIRT1 protein and mRNA was down-regulated, and the methylation of RNA m6A was increased, the novel object recognition index was increased, the expression of METTL3 was down-regulated, the expression of SIRT1 protein and mRNA was up-regulated, and the methylation of RNA m6A was decreased in SP+ HSR group( P<0.05). Compared to SP+ HSR group, the escape latency was significantly prolonged at 2-6 days, the time spent in the target quadrant was shortened, the number of crossing the original platform was decreased, the novel object recognition index was decreased, the expression of METTL3 was up-regulated, the expression of SIRT1 protein and mRNA was down-regulated, and the methylation of RNA m6A was increased in METTL3+ SP+ HSR group( P<0.05), and no significant change was found in the aforementioned indicators in NC+ SP+ HSR group ( P>0.05). Conclusions:The mechanism by which sevoflurane post-conditioning alleviates cognitive dysfunction is associated with down-regulation of METTL3 expression, reduction of RNA m6A methylation, and up-regulation of SIRT1 expression in HSR mice.
9.Validity and reliability of the Chinese version of children’s trust in general nurses scale
Qingqing CHONG ; Dan CHEN ; Zhiheng ZHAN ; Jing CHEN ; Enfang SHAN ; Xianwen LI
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(11):1032-1037
Objective:To translate the Children's trust in general nurses scale (CTGNS) into Chinese, and conduct item response theory (IRT) analysis and reliability and validity evaluation.Methods:Following Brislin's translation model, the CTGNS was translated, back-translated, and cross-culturally adapted to form the Chinese version of the CTGNS.A total of 502 pediatric inpatients from a pediatric hospital in Nanjing city were surveyed from July to August 2022.The Samejima graded response model in IRT was used to calculate the difficulty and discrimination parameters of each item, while item characteristic curves were plotted.The reliability and validity of the Chinese version CTGNS were evaluated.SPSS 26.0, Mplus 8.0 and MULTILOG 7.03 softwares were used for data analysis.Results:The item discrimination of the Chinese version CTGNS ranged from 0.088 to 2.730.The difficulty parameters ranged from -4.667 to 26.094 and increased with increasing difficulty levels without any reverse thresholds.The item characteristic curves exhibited sound shapes for all items except for item 9.Based on the selection criteria of the corresponding parameters, 8 items were retained.The revised Chinese version CTGNS (8 items) consisted of 3 dimensions.The content validity index (CVI) of each item ranged from 0.833 to 1.000, and the total scale CVI was 0.960.The Cronbach's α coefficient for the overall scale was 0.822, and the Cronbach's α coefficients for each dimension ranged from 0.752 to 0.852.The test-retest reliability of the scale was 0.87.Exploratory factor analysis yielded 3 common factors with a cumulative variance contribution rate of 73.468%.Confirmatory factor analysis indicated a good model fit( χ2/ df=2.674, RMSEA=0.082, SRMR=0.046, CFI=0.964). Conclusion:The revised Chinese version CTGNS demonstrates sound reliability and validity and can be used as an assessment tool for measuring children's trust in nurses.
10.Role of frontal lobe and its related circuits involved in cognitive flexibility impairment in autism
Chengming XU ; Yalei FAN ; Zhe ZHANG ; Jiwei ZHANG ; Liguo LI ; Xianwen DONG
Chinese Journal of Behavioral Medicine and Brain Science 2023;32(11):1051-1056
Autism spectrum disorder (ASD) is a multifactorial disease, with social difficulties and repetitive behaviors as its core symptoms. With the improvement of diagnostic methods, the detection rate of ASD is increasing year by year.Cognitive flexibility impairment is very obvious in most autistic patients.More and more studies have shown that cognitive flexibility impairment is related to the occurrence and development of core symptoms. However, the mechanism of cognitive flexibility impairment in autism remains unclear. The frontal lobe plays an important role in advanced cognition, and its complete development is related to cognitive function. Recent studies have shown that frontal lobe dysfunction is closely related to cognitive flexibility deficits in autistic patients, and the abnormal changes in the frontal lobe, the associated default mode network dysfunction and frontal striatal circuit defects may be the important mechanisms of cognitive flexibility impairment. Based on the recent clinical and basic studies on cognitive flexibility in autism, this article reviews the mechanisms of frontal lobe and related circuits involved in the impairment of cognitive flexibility in autism.


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