1.Therapeutic Effects of Wumei Wan and Its Disassembled Prescriptions on Diabetic Gastroparesis Model Mice and Influence on the Expression of Oxidative Stress Factors
Zi-Miao WANG ; Yi CHEN ; Yue YAN ; Shi-Jian QUAN
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(10):2778-2787
		                        		
		                        			
		                        			Objective To observe the therapeutic effect and compatibility mechanism of Wumei Wan on diabetic gastroparesis(DGP)mice.Methods Sixty-nine C57BL/6J mice were randomly divided into normal group(12 mice)and model group(57 mice).The DGP model was constructed by 60%high-fat diet combined with intraperitoneal injection of streptozotocin(STZ).After successful modeling,the mice in the model group were randomly divided into model group,Wumei Wan whole prescription group,Wumei Wan sour medicine group,Wumei Wan bitter medicine group,Wumei Wan sweet medicine group and Wumei Wan pungent medicine group,with nine mice in each group.After 28 days of treatment,the gastric emptying rate and small intestinal propulsion rate of mice were measured.The pathological changes of gastric antrum were observed by hematoxylin-eosin(HE)staining.The fasting blood glucose(FBS)of mice was detected by blood glucose meter.The levels of triglyceride(TG),total cholesterol(TC)and low density lipoprotein cholesterol(LDL-C)were detected by colorimetry.The contents of gastrin(GAS),motilin(MTL)and vasoactive intestinal peptide(VIP)in gastric antrum were detected by enzyme-linked immunosorbent assay(ELISA).Western Blot was used to detect the expression of Kelch-like ECH-associated protein 1(Keap-1),nuclear factor erythroid 2-related factor 2(Nrf-2),NAD(P)H:quinone oxidoreductase 1(Nqo-1)and superoxide dismutase 1(Sod-1)in gastric antrum tissue.Results(1)Compared with the normal group,the levels of serum FBS,TG,TC and LDL-C in the model group were significantly increased(P<0.05 or P<0.01 or P<0.001);compared with the model group,the blood glucose level of the whole prescription group,the sour medicine group and the bitter medicine group was decreased significantly at the end of the third and fourth week(P<0.05 or P<0.01 or P<0.001),the serum TG,TC and LDL-C levels of the whole prescription group were significantly decreased(P<0.05 or P<0.01),the serum TC level of the sour medicine group,the bitter medicine group,the sweet medicine group and the pungent medicine group was significantly decreased(P<0.05),and only the LDL-C level of the sour medicine group was significantly decreased(P<0.05).(2)Compared with the normal group,the glandular cells in the gastric antrum mucosa and submucosa of the model group were disordered;compared with the model group,the pathological damage of gastric antrum tissue in Wumei Wan whole prescription group was significantly improved,the recovery effect of gastric antrum tissue damage in DGP mice in the sour medicine group and the bitter medicine group was superior to that in the sweet medicine group and the pungent medicine group.(3)Compared with the normal group,the gastric emptying rate and small intestinal propulsion rate of the model group were decreased(P<0.01 or P<0.001);compared with the model group,the gastric emptying rate of the whole prescription group,the sour medicine group and the bitter medicine group was significantly increased(P<0.05 or P<0.001),and the intestinal propulsion rate of the whole prescription group,the sour medicine group,the bitter medicine group,the sweet medicine group and the pungent medicine group was significantly increased(P<0.05 or P<0.01 or P<0.001).(4)Compared with the normal group,the content of VIP in the gastric antrum tissue of the model group was significantly increased(P<0.01),and the contents of GAS and MTL were significantly decreased(P<0.01 or P<0.001).Compared with the model group,the content of GAS in Wumei Wan whole prescription group,the sour medicine group,the bitter medicine group and the sweet medicine group was significantly increased(P<0.01 or P<0.001),the content of MTL in the whole prescription group and the bitter medicine group was significantly increased(P<0.05 or P<0.01),and the content of VIP in gastric antrum tissue of mice in Wumei Wan whole prescription group,sour medicine group,bitter medicine group and sweet medicine group was significantly decreased(P<0.05 or P<0.01 or P<0.001).(5)Compared with the normal group,the expression level of Keap-1 in the gastric antrum tissue of the model group was increased(P<0.05),and the expressions of Nrf-2,Nqo-1 and Sod-1 were decreased(P<0.05 or P<0.001).Compared with the model group,the expression level of Keap-1 protein in the whole prescription group and the bitter medicine group was significantly decreased(P<0.05 or P<0.001),and the expression levels of Nrf-2,Nqo-1 and Sod-1 in the whole prescription group,the sour medicine group and the bitter medicine group were significantly increased(P<0.05 or P<0.001).Conclusion Wumei Wan can effectively regulate glucose and lipid metabolism,improve gastric antrum injury and promote gastrointestinal motility in DGP mice.The efficacy of the whole prescription is superior to that of each disassembled prescription.The mechanism may be related to the synergistic compatibility of sour medicine,bitter medicine,sweet medicine and pungent medicine,and mainly through sour medicine and bitter medicine regulating Nrf-2/Keap-1 pathway further to improve oxidative stress injury.
		                        		
		                        		
		                        		
		                        	
2.Synthesis and Single-cell Analysis Application of Antibody-Element-Tag Based on Peptide Affinity-mediated Site-specific Labeling Strategy
Lei WANG ; Li YI ; Hua-Min LI ; Shi CHEN ; Li-Min YANG ; Xiao-Wen YAN ; Qiu-Quan WANG
Chinese Journal of Analytical Chemistry 2024;52(10):1496-1507
		                        		
		                        			
		                        			Recently,inductively coupled plasma mass spectrometry(ICP-MS)has shown great potential in the quantitative analysis of biomolecules due to its high sensitivity,high accuracy,anti-interference ability and multi-element detection capability.Elemental tags enable specific labeling of target biomolecules with element-reporter groups,playing a critical role in the performance of bioanalysis using ICP-MS.Antibodies are highly specific to antigens on cell surfaces and are one of the most commonly used targeted delivery vehicles.Traditional random chemical coupling methods are difficult to control the coupling sites and loading numbers of element reporters,which lead to heterogeneity of antibody conjugates,affecting the specific recognition and binding ability of antibodies and antigens,and thus reducing the targeting ability and quantitative accuracy.To solve these issues,site-specific coupling of antibodies with element reporter was studied in this work.By using peptide-mediated affinity labeling technology,a short chain DNA was specifically labeled to immunoglobulin G(IgG)antibody K248,and then DNA base complementary pairing was used to realize accurate assembly of monoclonal antibodies and Eu-modified phage MS2 capsid,resulting in the preparation of highly sensitive and selective element tags.This method overcame the heterogeneity problem of element tags caused by random coupling,and ensured the targeting specificity,affinity and signal amplification ability of element tags.The element tag prepared here showed a specificity to bind/label with epidermal growth factor receptor(EGFR)biomarker on the surface of cancer cells,realizing single cancer cell analysis using ICP-MS.
		                        		
		                        		
		                        		
		                        	
3.Restoration-oriented anatomical analysis of alveolar bone at mandibular first molars and implications for immediate implant placement surgery: a CBCT study
Quan SHI ; Yang HUANG ; Na HUO ; Yi JIANG ; Tong ZHANG ; Juncheng WANG
The Journal of Advanced Prosthodontics 2024;16(4):212-220
		                        		
		                        			 PURPOSE:
		                        			This cone-beam computed tomography (CBCT) study aimed to analyze the anatomical characteristics of alveolar bone at mandibular first molar (MFM) and their implications for immediate implant placement surgery. 
		                        		
		                        			MATERIALS AND METHODS:
		                        			100 patients with 140 MFMs were reviewed retrospectively. We first performed a 3D reconstruction of the patient’s CBCT data to determine a reference plane with ideal implant placement and orientation. The following parameters of MFM region were analyzed: mesial-distal socket size (MDSS), buccal-lingual socket size (BL-SS), root furcation fornix to inferior alveolar nerve (IAN) distance (RF-I), interradicular bone thickness (IRB), mesial/distal root apex to the IAN distance (MRA-I/DRA-I), thickness of the buccal/lingual bone of the mesial root (MR-B/MR-L), thickness of the buccal/lingual bone of the distal root (DR-B/DR-L).  
		                        		
		                        			RESULTS:
		                        			The MD-SS of MFM was 8.74 ± 0.76 mm, and the BLSS was 8.26 ± 0.72 mm. The MR-B, DR-B was 1.01 ± 0.40 mm and 1.14 ± 0.50 mm, and the difference was statistically significant (P = .001). The values of the MR-L, DR-L were 2.71 ± 0.78 mm and 3.09 ± 0.73 mm, and the difference was also statistically significant (P < .001). The mean distance of RF-I was 15.68 ± 2.13mm, and the MRA-I was 7.06 ± 2.22 mm, which was greater than that of DRA-I (6.48 ± 2.30 mm, P < .001). The IRB at 2 mm, 4 mm apical from the furcation fornix, and at apex level was 2.81 ± 0.50 mm, 3.30 ± 0.62 mm, and 4.44 ± 1.02 mm, respectively.  
		                        		
		                        			CONCLUSION
		                        			There is relatively sufficient bone mass in interradicular bone in height, but an adequate width is lacking for the bone between the mesial and distal root after the extraction of the MFM for immediate implantation. The thickness of the MFM buccal bone is relative thin, especially for the mesial root. 
		                        		
		                        		
		                        		
		                        	
4.Risk factors for neonatal asphyxia and establishment of a nomogram model for predicting neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture: a multicenter study.
Fang JIN ; Yu CHEN ; Yi-Xun LIU ; Su-Ying WU ; Chao-Ce FANG ; Yong-Fang ZHANG ; Lu ZHENG ; Li-Fang ZHANG ; Xiao-Dong SONG ; Hong XIA ; Er-Ming CHEN ; Xiao-Qin RAO ; Guang-Quan CHEN ; Qiong YI ; Yan HU ; Lang JIANG ; Jing LI ; Qing-Wei PANG ; Chong YOU ; Bi-Xia CHENG ; Zhang-Hua TAN ; Ya-Juan TAN ; Ding ZHANG ; Tie-Sheng YU ; Jian RAO ; Yi-Dan LIANG ; Shi-Wen XIA
Chinese Journal of Contemporary Pediatrics 2023;25(7):697-704
		                        		
		                        			OBJECTIVES:
		                        			To investigate the risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture and establish a nomogram model for predicting the risk of neonatal asphyxia.
		                        		
		                        			METHODS:
		                        			A retrospective study was conducted with 613 cases of neonatal asphyxia treated in 20 cooperative hospitals in Enshi Tujia and Miao Autonomous Prefecture from January to December 2019 as the asphyxia group, and 988 randomly selected non-asphyxia neonates born and admitted to the neonatology department of these hospitals during the same period as the control group. Univariate and multivariate analyses were used to identify risk factors for neonatal asphyxia. R software (4.2.2) was used to establish a nomogram model. Receiver operator characteristic curve, calibration curve, and decision curve analysis were used to assess the discrimination, calibration, and clinical usefulness of the model for predicting the risk of neonatal asphyxia, respectively.
		                        		
		                        			RESULTS:
		                        			Multivariate logistic regression analysis showed that minority (Tujia), male sex, premature birth, congenital malformations, abnormal fetal position, intrauterine distress, maternal occupation as a farmer, education level below high school, fewer than 9 prenatal check-ups, threatened abortion, abnormal umbilical cord, abnormal amniotic fluid, placenta previa, abruptio placentae, emergency caesarean section, and assisted delivery were independent risk factors for neonatal asphyxia (P<0.05). The area under the curve of the model for predicting the risk of neonatal asphyxia based on these risk factors was 0.748 (95%CI: 0.723-0.772). The calibration curve indicated high accuracy of the model for predicting the risk of neonatal asphyxia. The decision curve analysis showed that the model could provide a higher net benefit for neonates at risk of asphyxia.
		                        		
		                        			CONCLUSIONS
		                        			The risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture are multifactorial, and the nomogram model based on these factors has good value in predicting the risk of neonatal asphyxia, which can help clinicians identify neonates at high risk of asphyxia early, and reduce the incidence of neonatal asphyxia.
		                        		
		                        		
		                        		
		                        			Infant, Newborn
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Pregnancy
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Nomograms
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Cesarean Section
		                        			;
		                        		
		                        			Risk Factors
		                        			;
		                        		
		                        			Asphyxia Neonatorum/etiology*
		                        			
		                        		
		                        	
5.A novel method for electroencephalography background analysis in neonates with hypoxic-ischemic encephalopathy.
Xiu-Ying FANG ; Yi-Li TIAN ; Shu-Yuan CHEN ; Quan SHI ; Duo ZHENG ; Ying-Jie WANG ; Jian MAO
Chinese Journal of Contemporary Pediatrics 2023;25(2):128-134
		                        		
		                        			OBJECTIVES:
		                        			To explore a new method for electroencephalography (EEG) background analysis in neonates with hypoxic-ischemic encephalopathy (HIE) and its relationship with clinical grading and head magnetic resonance imaging (MRI) grading.
		                        		
		                        			METHODS:
		                        			A retrospective analysis was performed for the video electroencephalography (vEEG) and amplitude-integrated electroencephalography (aEEG) monitoring data within 24 hours after birth of neonates diagnosed with HIE from January 2016 to August 2022. All items of EEG background analysis were enrolled into an assessment system and were scored according to severity to obtain the total EEG score. The correlations of total EEG score with total MRI score and total Sarnat score (TSS, used to evaluate clinical gradings) were analyzed by Spearman correlation analysis. The total EEG score was compared among the neonates with different clinical gradings and among the neonates with different head MRI gradings. The receiver operating characteristic (ROC) curve and the area under thecurve (AUC) were used to evaluate the value of total EEG score in diagnosing moderate/severe head MRI abnormalities and clinical moderate/severe HIE, which was then compared with the aEEG grading method.
		                        		
		                        			RESULTS:
		                        			A total of 50 neonates with HIE were included. The total EEG score was positively correlated with the total head MRI score and TSS (rs=0.840 and 0.611 respectively, P<0.001). There were significant differences in the total EEG score between different clinical grading groups and different head MRI grading groups (P<0.05). The total EEG score and the aEEG grading method had an AUC of 0.936 and 0.617 respectively in judging moderate/severe head MRI abnormalities (P<0.01) and an AUC of 0.887 and 0.796 respectively in judging clinical moderate/severe HIE (P>0.05). The total EEG scores of ≤6 points, 7-13 points, and ≥14 points were defined as mild, moderate, and severe EEG abnormalities respectively, which had the best consistency with clinical grading and head MRI grading (P<0.05).
		                        		
		                        			CONCLUSIONS
		                        			The new EEG background scoring method can quantitatively reflect the severity of brain injury and can be used for the judgment of brain function in neonates with HIE.
		                        		
		                        		
		                        		
		                        			Infant, Newborn
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Hypoxia-Ischemia, Brain/diagnostic imaging*
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Brain Injuries
		                        			;
		                        		
		                        			Electroencephalography
		                        			;
		                        		
		                        			ROC Curve
		                        			
		                        		
		                        	
6.Systematic review and Meta-analysis of efficacy and safety of Shufeng Jiedu Capsules in treatment of influenza.
Feng ZHOU ; Guo-Zhen ZHAO ; Bo LI ; Xiao-Long XU ; Yi-Fan SHI ; Yi-Yi MAO ; Jin-Hao TIAN ; Qing-Quan LIU
China Journal of Chinese Materia Medica 2023;48(22):6216-6224
		                        		
		                        			
		                        			This study aims to systematically review the efficacy and safety of Shufeng Jiedu Capsules in the treatment of influenza. The randomized controlled trial(RCT) of Shufeng Jiedu Capsules alone or in combination with conventional western medicine for treating influenza were retrieved from PubMed, EMbase, Cochrane Library, Web of Science, SinoMed, CNKI, VIP, Wanfang, and ClinicalTrails.gov. The data analysis was performed in RevMan 5.4.1. The Cochrane risk of bias assessment tool was used to evaluate the quality of the involved RCT, and GRADEpro GDT to assess the quality of the evidence. A total of 11 RCTs involving 1 836 patients were included in this study. Compared with conventional western medicine, Shufeng Jiedu Capsules/Shufeng Jiedu Capsules + conventional western medicine improved the response rate(RR=1.09, 95%CI[1.03, 1.15], P=0.002), shortened the time to relief of cough, and increased the 3-day sore throat relief rate, whereas there was no significant difference in the time to fever abatement, the time to relief of sore throat, 3-day cough relief rate, or 3-day runny nose relief rate. Subgroup-analysis showed that Shufeng Jiedu Capsules + conventional western medicine improved the response rate(RR=1.11, 95%CI[1.08, 1.15], P<0.000 01), shortened the time to relief of cough, and increased the 3-day relief rate of symptoms(cough, sore throat, and runny nose) compared with conventional western medicine alone, while there was no significant difference in the time to fever abatement or the time to relief of sore throat. Shufeng Jiedu Capsules alone could not improve the response rate(RR=0.97, 95%CI[0.93, 1.02], P=0.19). In addition, Shufeng Jiedu Capsules/Shufeng Jiedu Capsules + conventional western medicine vs conventional western medicine were no significant difference in adverse reactions(RR=0.98, 95%CI[0.57, 1.69], P=0.95). The available evidence suggests that Shufeng Jiedu Capsules is effective and safe in the treatment of influenza, and the combination of Shufeng Jiedu Capsules with conventional western medicine can accelerate the relief of symptoms. However, since the number and quality of the included studies were low, the above findings remained to be further verified by multicenter RCT with large sample sizes.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Influenza, Human/drug therapy*
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/adverse effects*
		                        			;
		                        		
		                        			Capsules
		                        			;
		                        		
		                        			Cough/chemically induced*
		                        			;
		                        		
		                        			Pharyngitis
		                        			;
		                        		
		                        			Rhinorrhea
		                        			;
		                        		
		                        			Multicenter Studies as Topic
		                        			
		                        		
		                        	
7.Schisandrin B Protects against Ischemic Brain Damage by Regulating PI3K/AKT Signaling in Rats.
Quan-Long HONG ; Yi-Hang DING ; Jing-Yi CHEN ; Song-Sheng SHI ; Ri-Sheng LIANG ; Xian-Kun TU
Chinese journal of integrative medicine 2023;29(10):885-894
		                        		
		                        			OBJECTIVE:
		                        			To explore the effect and mechanism of schisandrin B (Sch B) in the treatment of cerebral ischemia in rats.
		                        		
		                        			METHODS:
		                        			The cerebral ischemia models were induced by middle cerebral artery occlusion (MCAO) and reperfusion. Sprague-Dawley rats were divided into 6 groups using a random number table, including sham, MCAO, MCAO+Sch B (50 mg/kg), MCAO+Sch B (100 mg/kg), MCAO+Sch B (100 mg/kg)+LY294002, and MCAO+Sch B (100 mg/kg)+wortmannin groups. The effects of Sch B on pathological indicators, including neurological deficit scores, cerebral infarct volume, and brain edema, were subsequently studied. Tissue apoptosis was identified by terminal transferase-mediated dUTP nick end-labeling (TUNEL) staining. The protein expressions involved in apoptosis, inflammation response and oxidative stress were examined by immunofluorescent staining, biochemical analysis and Western blot analysis, respectively. The effect of Sch B on phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling was also explored.
		                        		
		                        			RESULTS:
		                        			Sch B treatment decreased neurological deficit scores, cerebral water content, and infarct volume in MCAO rats (P<0.05 or P<0.01). Neuronal nuclei and TUNEL staining indicated that Sch B also reduced apoptosis in brain tissues, as well as the Bax/Bcl-2 ratio and caspase-3 expression (P<0.01). Sch B regulated the production of myeloperoxidase, malondialdehyde, nitric oxide and superoxide dismutase, as well as the release of cytokine interleukin (IL)-1 β and IL-18, in MCAO rats (P<0.05 or P<0.01). Sch B promoted the phosphorylation of PI3K and AKT. Blocking the PI3K/AKT signaling pathway with LY294002 or wortmannin reduced the protective effect of Sch B against cerebral ischemia (P<0.05 or P<0.01).
		                        		
		                        			CONCLUSIONS
		                        			Sch B reduced apoptosis, inflammatory response, and oxidative stress of MCAO rats by modulating the PI3K/AKT pathway. Sch B had a potential for treating cerebral ischemia.
		                        		
		                        		
		                        		
		                        	
8.Polypeptide from Moschus Suppresses Lipopolysaccharide-Induced Inflammation by Inhibiting NF-κ B-ROS/NLRP3 Pathway.
Jing YI ; Li LI ; Zhu-Jun YIN ; Yun-Yun QUAN ; Rui-Rong TAN ; Shi-Long CHEN ; Ji-Rui LANG ; Jiao LI ; Jin ZENG ; Yong LI ; Zi-Jian SUN ; Jun-Ning ZHAO
Chinese journal of integrative medicine 2023;29(10):895-904
		                        		
		                        			OBJECTIVE:
		                        			To examine the anti-inflammatory effects and potential mechanisms of polypeptide from Moschus (PPM) in lipopolysaccharide (LPS)-induced THP-1 macrophages and BALB/c mice.
		                        		
		                        			METHODS:
		                        			The polypeptide was extracted from Moschus and analyzed by high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Subsequently, LPS was used to induce inflammation in THP-1 macrophages and BALB/c mice. In LPS-treated or untreated THP-1 macrophages, cell viability was observed by cell counting kit 8 and lactate dehydrogenase release assays; the proinflammatory cytokines and reactive oxygen species (ROS) were measured by enzyme-linked immunosorbent assay and flow cytometry, respectively; and protein and mRNA levels were measured by Western blot and real-time quantitative polymerase chain reaction (qRT-PCR), respectively. In LPS-induced BALB/c mice, the proinflammatory cytokines were measured, and lung histology and cytokines were observed by hematoxylin and eosin (HE) and immunohistochemical (IHC) staining, respectively.
		                        		
		                        			RESULTS:
		                        			The SDS-PAGE results suggested that the molecular weight of purified PPM was in the range of 10-26 kD. In vitro, PPM reduced the production of interleukin 1β (IL-1β), IL-18, tumor necrosis factor α (TNF-α), IL-6 and ROS in LPS-induced THP-1 macrophages (P<0.01). Western blot analysis demonstrated that PPM inhibited LPS-induced nuclear factor κB (NF-κB) pathway and thioredoxin interacting protein (TXNIP)/nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain containing 3 (NLRP3) inflammasome pathway by reducing protein expression of phospho-NF-κB p65, phospho-inhibitors of NF-κB (Iκ Bs) kinase α/β (IKKα/β), TXNIP, NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC), and pro-caspase-1 (P<0.05 or P<0.01). In addition, qRT-PCR revealed the inhibitory effects of PPM on the mRNA levels of TXNIP, NLRP3, ASC, and caspase-1 (P<0.05 or P<0.01). Furthermore, in LPS-induced BALB/c mice, PPM reduced TNF-α and IL-6 levels in serum (P<0.05 or P<0.01), decreased IL-1β and IL-18 levels in the lungs (P<0.01) and alleviated pathological injury to the lungs.
		                        		
		                        			CONCLUSION
		                        			PPM could attenuate LPS-induced inflammation by inhibiting the NF-κB-ROS/NLRP3 pathway, and may be a novel potential candidate drug for treating inflammation and inflammation-related diseases.
		                        		
		                        		
		                        		
		                        	
9.Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification.
Sun-Xiang WEI ; Hui-Xiang CHEN ; Sheng HU ; Yi-Xia ZHAO ; Hui-Xia SHI ; Zhe WANG ; Wen LI ; An-Quan JI ; Qi-Fan SUN
Journal of Forensic Medicine 2022;38(6):719-725
		                        		
		                        			OBJECTIVES:
		                        			To establish a system for simultaneous detection of miR-888 and miR-891a by droplet digital PCR (ddPCR), and to evaluate its application value in semen identification.
		                        		
		                        			METHODS:
		                        			The hydrolysis probes with different fluorescence modified reporter groups were designed to realize the detection of miR-888 and miR-891a by duplex ddPCR. A total of 75 samples of 5 body fluids (including peripheral blood, menstrual blood, semen, saliva and vaginal secretion) were detected. The difference analysis was conducted by Mann-Whitney U test. The semen differentiation ability of miR-888 and miR-891a was evaluated by ROC curve analysis and the optimal cut-off value was obtained.
		                        		
		                        			RESULTS:
		                        			There was no significant difference between the dual-plex assay and the single assay in this system. The detection sensitivity was up to 0.1 ng total RNA, and the intra- and inter-batch coefficients of variation were less than 15%. The expression levels of miR-888 and miR-891a detected by duplex ddPCR in semen were both higher than those in other body fluids. ROC curve analysis showed that the AUC of miR-888 was 0.976, the optimal cut-off value was 2.250 copies/μL, and the discrimination accuracy was 97.33%; the AUC of miR-891a was 1.000, the optimal cut-off value was 1.100 copies/μL, and the discrimination accuracy was 100%.
		                        		
		                        			CONCLUSIONS
		                        			In this study, a method for detection of miR-888 and miR-891a by duplex ddPCR was successfully established. The system has good stability and repeatability and can be used for semen identification. Both miR-888 and miR-891a have high ability to identify semen, and the discrimination accuracy of miR-891a is higher.
		                        		
		                        		
		                        		
		                        			Female
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Body Fluids/chemistry*
		                        			;
		                        		
		                        			MicroRNAs/analysis*
		                        			;
		                        		
		                        			Real-Time Polymerase Chain Reaction/methods*
		                        			;
		                        		
		                        			Saliva/chemistry*
		                        			;
		                        		
		                        			Semen/chemistry*
		                        			;
		                        		
		                        			Male
		                        			
		                        		
		                        	
10.One-year follow-up results of atrial fibrillation patients who undergoing transcatheter aortic valve implantation.
Shi Chen ZHOU ; Kai XU ; Bin WANG ; Geng WANG ; Zhen Yang LIANG ; Yang LI ; Yi FANG ; Ling Fei ZHENG ; Yan Qiu WANG ; Wei Wei ZHOU ; Quan Min JING ; Ya Ling HAN
Chinese Journal of Cardiology 2022;50(2):132-136
		                        		
		                        			
		                        			Objective: To investigate whether atrial fibrillation (AF) before transcatheter aortic valve implantation (TAVI) will affect the prognosis of patients post TAVI. Methods: This is a single center retrospective study. A total of 115 patients with severe aortic stenosis (AS) who were admitted to General Hospital of Northern Theater Command from May 2016 to November 2020 and successfully received TAVI treatment were included. According to absence or accompaniment of AF pre-TAVI, they were divided into AF group (21 cases) and non-AF group (94 cases). The patients were followed up for postoperative antithrombotic treatment and the occurrence of the net adverse clinical and cerebrovascular events (NACCE) at 12 months post TAVI, including cardiogenic death, readmission to hospital for heart failure, nonfatal myocardial infarction, ischemic stroke and severe bleeding (BARC levels 3-5). Univariate logistic regression was used to analyze the related factors of NACCE. Results: Among the 115 selected patients, age was (73.8±6.9) years, there were 63 males. And 21 cases (18.2%) were diagnosed as AFbefore TAVI. In terms of postoperative antithrombotic therapy, 48.9% (46/94) of the patients in the non-AF group received monotherapy and 47.9% (45/94) received dual antiplatelet therapy. In the AF group, 47.6% (10/21) received anticoagulants and 33.3% (7/21) received dual antiplatelet therapy. The proportion of patients in the AF group taking non-vitamin K antagonist oral anticoagulants (NOAC) was higher than that in the non-AF group (38.1% (8/21) vs. 2.1% (2/94), P<0.001). Patients in both groups were followed up to 12 months after TAVI. During the 12 months follow-up, the incidence of NACCE after TAVI was 14.3% (3/21) in the AF group, which was numerically higher than that in the non-AF group (6.4% (6/94)), but the difference was not statistically significant (P=0.441). The incidence of severe bleeding was significantly higher in the AF group than in the non-AF group (9.5% (2/21) vs. 0, P=0.032). Univariate logistic regression analysis showed that hypertension was associated with the risk of NACCE (OR=8.308, P=0.050), while AF was not associated with the risk of NACCE (P=0.235). Conclusion: The incidence of severe bleeding after TAVI is higher in patients with AF than in patients without AF prior TAVI, and there is a trend of increased risk of NACCE post TAVI in AF patients.
		                        		
		                        		
		                        		
		                        			Aged
		                        			;
		                        		
		                        			Aged, 80 and over
		                        			;
		                        		
		                        			Anticoagulants
		                        			;
		                        		
		                        			Aortic Valve
		                        			;
		                        		
		                        			Aortic Valve Stenosis/surgery*
		                        			;
		                        		
		                        			Atrial Fibrillation/drug therapy*
		                        			;
		                        		
		                        			Follow-Up Studies
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Risk Factors
		                        			;
		                        		
		                        			Transcatheter Aortic Valve Replacement
		                        			;
		                        		
		                        			Treatment Outcome
		                        			
		                        		
		                        	
            
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