1.Construction of nursing teaching rounds model in dental clinics based on constructivism learning theory
Siyuan MOU ; Lin LIU ; Shouhui CHEN ; Yingqian QIU
Chinese Journal of Medical Education Research 2024;23(5):702-707
Objective:To construct a model of nursing teaching rounds in dental clinics, and to provide a reference for the implementation of nursing teaching activities in dental clinics.Methods:Using the constructivism learning theory framework, we developed a preliminary draft of a teaching rounds model for dental outpatient nursing according to literature review, cross-sectional survey, and brainstorming results. Two rounds of Delphi consultation with 21 experts from across China were conducted to determine the teaching rounds model for dental outpatient nursing. Data entry and statistical analysis were performed with Excel 2016 and SPSS 26.0 software.Results:In the two rounds of expert consultation, the questionnaire response rates were both 100%; the expert authority coefficients were 0.865 and 0.873, respectively; and the Kendall's coefficients of concordance were 0.116 and 0.164, respectively. The established dental outpatient nursing teaching rounds model included 7 first-level items and 22 second-level items.Conclusions:The dental outpatient nursing teaching rounds model constructed in this study is scientific, reliable, and practical, which can provide guidance for conducting nursing teaching rounds in dental clinics.
2.Effects of gelatin methacrylate anhydride hydrogel loaded with small extracellular vesicles derived from human umbilical cord mesenchymal stem cells in the treatment of full-thickness skin defect wounds in mice
Yiqi CHEN ; Yingqian ZHOU ; Qian WEI ; Xiaoye XIE ; Xinzhu LIU ; Dawei LI ; Chuan'an SHEN
Chinese Journal of Burns 2024;40(4):323-332
Objective:To investigate the effects of gelatin methacrylate anhydride (GelMA) hydrogel loaded with small extracellular vesicles derived from human umbilical cord mesenchymal stem cells (hUCMSCs-sEVs) in the treatment of full-thickness skin defect wounds in mice.Methods:This study was an experimental study. hUCMSCs-sEVs were extracted by ultracentrifugation, their morphology was observed through transmission electron microscope, and the expression of CD9, CD63, tumor susceptibility gene 101 (TSG101), and calnexin was detected by Western blotting. The human umbilical vein endothelial cells (HUVECs), the 3 rd and 4 th passages of human epidermal keratinocytes (HEKs) and human dermal fibroblasts (HDFs) were all divided into blank control group (routinely cultured) and hUCMSC-sEV group (cultured with the cell supernatant containing hUCMSCs-sEVs). The cell scratch test was performed and the cell migration rates at 6, 12, and 24 h after scratching were calculated, the cell Transwell assay was performed and the number of migration cells at 12 h after culture was calculated, and the proportion of proliferating cells was detected by 5-acetylidene-2'-deoxyuridine and Hoechst staining at 24 h after culture, with sample numbers being all 3. The simple GelMA hydrogel and the GelMA hydrogel loaded with hUCMSCs-sEVs (hereinafter referred to as hUCMSC-sEV/GelMA hydrogel) were prepared. Then the micromorphology of 2 kinds of hydrogels was observed under scanning electron microscope, the distribution of hUCMSCs-sEVs was observed by laser scanning confocal microscope, and the cumulative release rates of hUCMSCs-sEVs at 0 (immediately), 2, 4, 6, 8, 10, and 12 d after soaking hUCMSC-sEV/GelMA hydrogel in phosphate buffer solution (PBS) were measured and calculated by protein colorimetric quantification ( n=3). Twenty-four 6-week-old male C57BL/6J mice were divided into PBS group, hUCMSC-sEV alone group, GelMA hydrogel alone group, and hUCMSC-sEV/GelMA hydrogel group according to the random number table, with 6 mice in each group, and after the full-thickness skin defect wounds on the back of mice in each group were produced, the wounds were performed with PBS injection, hUCMSC-sEV suspenson injection, simple GelMA coverage, and hUCMSC-sEV/GelMA hydrogel coverage, respectively. Wound healing was observed on post injury day (PID) 0 (immediately), 4, 8, and 12, and the wound healing rates on PID 4, 8, and 12 were calculated, and the wound tissue was collected on PID 12 for hematoxylin-eosin staining to observe the structure of new tissue, with sample numbers being both 6. Results:The extracted hUCMSCs-sEVs showed a cup-shaped structure and expressed CD9, CD63, and TSG101, but barely expressed calnexin. At 6, 12, and 24 h after scratching, the migration rates of HEKs (with t values of 25.94, 20.98, and 20.04, respectively), HDFs (with t values of 3.18, 5.68, and 4.28, respectively), and HUVECs (with t values of 4.32, 19.33, and 4.00, respectively) in hUCMSC-sEV group were significantly higher than those in blank control group ( P<0.05). At 12 h after culture, the numbers of migrated HEKs, HDFs, and HUVECs in hUCMSC-sEV group were 550 ±23, 235 ±9, and 856 ±35, respectively, which were significantly higher than 188 ±14, 97 ±6, and 370 ±32 in blank control group (with t values of 22.95, 23.13, and 17.84, respectively , P<0.05). At 24 h after culture, the proportions of proliferating cells of HEKs, HDFs, and HUVECs in hUCMSC-sEV group were significantly higher than those in blank control group (with t values of 22.00, 13.82, and 32.32, respectively, P<0.05). The inside of simple GelMA hydrogel showed a loose and porous sponge-like structure, and hUCMSCs-sEVs was not observed in it. The hUCMSC-sEV/GelMA hydrogel had the same sponge-like structure, and hUCMSCs-sEVs were uniformly distributed in clumps. The cumulative release rate curve of hUCMSCs-sEVs from hUCMSC-sEV/GelMA hydrogel tended to plateau at 2 d after soaking, and the cumulative release rate of hUCMSCs-sEVs was (59.2±1.8)% at 12 d after soaking. From PID 0 to 12, the wound areas of mice in the 4 groups gradually decreased. On PID 4, 8, and 12, the wound healing rates of mice in hUCMSC-sEV/GelMA hydrogel group were significantly higher than those in the other 3 groups ( P<0.05); the wound healing rates of mice in GelMA hydrogel alone group and hUCMSC-sEV alone group were significantly higher than those in PBS group ( P<0.05). On PID 8 and 12, the wound healing rates of mice in hUCMSC-sEV alone group were significantly higher than those in GelMA hydrogel alone group ( P<0.05). On PID 12, the wounds of mice in hUCMSC-sEV/GelMA hydrogel group showed the best wound epithelization, loose and orderly arrangement of dermal collagen, and the least number of inflammatory cells, while the dense arrangement of dermal collagen and varying degrees of inflammatory cell infiltration were observed in the wounds of mice in the other 3 groups. Conclusions:hUCMSCs-sEVs can promote the migration and proliferation of HEKs, HDFs, and HUVECs which are related to skin wound healing, and slowly release in GelMA hydrogel. The hUCMSC-sEV/GelMA hydrogel as a wound dressing can significantly improve the healing speed of full-thickness skin defect wounds in mice.
3.Artificial intelligence for brain disease diagnosis using electroencephalogram signals
SHANG SHUNUO ; SHI YINGQIAN ; ZHANG YAJIE ; LIU MENGXUE ; ZHANG HONG ; WANG PING ; ZHUANG LIUJING
Journal of Zhejiang University. Science. B 2024;25(10):914-940
Brain signals refer to electrical signals or metabolic changes that occur as a consequence of brain cell activity.Among the various non-invasive measurement methods,electroencephalogram(EEG)stands out as a widely employed technique,providing valuable insights into brain patterns.The deviations observed in EEG reading serve as indicators of abnormal brain activity,which is associated with neurological diseases.Brain?computer interface(BCI)systems enable the direct extraction and transmission of information from the human brain,facilitating interaction with external devices.Notably,the emergence of artificial intelligence(AI)has had a profound impact on the enhancement of precision and accuracy in BCI technology,thereby broadening the scope of research in this field.AI techniques,encompassing machine learning(ML)and deep learning(DL)models,have demonstrated remarkable success in classifying and predicting various brain diseases.This comprehensive review investigates the application of AI in EEG-based brain disease diagnosis,highlighting advancements in AI algorithms.
4.Genetic safety evaluation of allogeneic bone marrow mesenchymal stem cells in hosts following traumatic brain injury
Sixian HUANG ; Zhiming FENG ; Yu XIE ; Xiaoxiong ZOU ; Kunlin LIU ; Shiting HUA ; Cong LI ; Yuxi ZOU ; Yingqian CAI ; Yanping TANG ; Xiaodan JIANG
Chinese Journal of Neuromedicine 2023;22(6):575-584
Objective:To investigate the genetic safety of allogeneic bone marrow mesenchymal stem cells (BMSCs) transplantation in traumatic brain injury (TBI).Methods:(1) In vivo experiment: BMSCs from male SD rats were isolated and cultured. Moderate TBI models were prepared by implanting and fixing micro-drug injection cannula into the left ventricle of 12 female SD rats, and 3 d after that, striking the right cerebral cortex of the rats with pneumatic precision percussion device was performed. Four h, and 3, 6, 9, and 12 d after modeling, TBI rats were given a single/multiple BMSCs infusion (2.5×10 5/time, total volume 10 μL) by cannula; 48 and 72 h, and 10 and 14 d after modeling, brain tissues of TBI rats (3 at each time point) were prepared into paraffin specimens. Immunofluorescent staining was used to detect the microglia activation, and RNAscope ? technology was used to detect the co-localization of astrocytes, neurons, microglia and transplanted BMSCs to observe whether the allogeneic BMSCs were integrated with the host brain cells after transplantation into TBI host. (2) In vitro experiment: the frozen and revived microglial cell line BV2 was transfected with green fluorescent protein (GFP)-positive lentiviral particles, and then, BMSCs prelabeled with pHrodo RED probe and BV2 cells pretreated with lipopolysaccharide were co-cultured in a certain ratio (BV2:BMSCs=1:1, 1:2, 2:1); after 36 and 72 h of co-culture, the phagocytosis between the 2 kinds of cells was observed under confocal fluorescence inverted microscope to observe the specific action forms of microglia on BMSCs. Results:(1) In vivo experiment: 48 and 72 h, and 10 and 14 d after modeling, no colocalization of transplanted BMSCs with astrocytes or neurons was found in paraffin sections of brain tissue in TBI rats; however, 10 and 14 d after modeling, microglia in TBI rats were obviously activated and migrated to the left lateral ventricle and choroid plexus, and co-localization of microglia with transplanted BMSCs was observed. (2) In vitro experiment: phagocytosis occurred after co-culture of BV2 cells at different proportions with BMSCs for 36 and 72 h. Conclusion:After transplantation, allogeneic BMSCs do not integrate with astrocytes or neurons of the TBI host, but they could be phagocytosed by microglia, indicating that allogeneic BMSCs transplantation for TBI is genetically safe.
5.Association between perfluorinated alkyl substances and depression
Jing LIU ; Lulu ZHANG ; Yingqian HE ; Yanan ZHAO ; Shan ZHENG ; Minzhen WANG
Journal of Environmental and Occupational Medicine 2023;40(6):688-694
Background Perfluorinated alkyl substances (PFAS) are a class of synthetic organic fluorides, which have adverse health effects on brain function, and limited research has been conducted on their effects on depression. Objective To assess potential correlation between serum PFAS and depression. Methods Using the 2015—2016 and 2017—2018 National Health and Nutrition Examination Survey (NHANES) datasets, 2626 subjects with complete relevant information in people ≥20 years old were selected. Logistic regression and restricted cubic splines were used to analyze the association and dose-response relationship between serum PFAS concentration and depression. Subgroup analysis was performed on sex, age, race, education level, marital status, family income to poverty ratio, moderate exercise, body mass index, and drinking status. Results Among the 2626 subjects, there were 666 patients (25.4%) with mild or above depression. After adjusting for race, education level, marital status, body mass index, moderate exercise, drinking history, cotinine, and other types of PFAS, serum perfluorooctane sulfonate (PFOS) was positively associated with the risk of depression (OR=1.85, 95%CI: 1.14, 3.02), and showed a nonlinear dose-response relationship (χ2=6.37, Pnonlinear=0.012). Perfluorononanoic acid (PFNA) was inversely associated with the risk of depression (OR=0.23, 95%CI: 0.14, 0.39), and showed a linear dose-response relationship (Ptrend<0.001, χ2=35.13, Poverall<0.001). After subgroup analysis, it was found that males, 20-39 year-olds and 40-64 year-olds were more sensitive to PFNA exposure (OR=0.15, 95%CI: 0.06, 0.37; OR=0.16, 95%CI: 0.06, 0.40; OR=0.18, 95%CI: 0.08, 0.39). PFOS only showed a statistically significant health effect in people aged 20-39 years (OR=3.00, 95%CI: 1.14, 7.94). In addition, among subgroups of non-Hispanic blacks, cohabitants, current drinkers, high school graduates, and obese patients, exposure to PFAS was significantly associated with the risk of depression. Conclusion PFOS exposure may be associated with increased levels of depression, whereas PFNA exposure may be protective.
6.Research progress on oral microbiota application in forensic medicine
Zhiyu GU ; Yunkun LIU ; Yijie CHEN ; Jiashuang LI ; Yingqian DUAN ; Xueqin SUN ; Yang LI
Chinese Journal of Forensic Medicine 2023;38(6):687-691
Forensic science is looking for clues at a crime scene in order to reconstruct the crime scene.Classic clues include DNA and fingerprints.Forensic microbiology is a branch of forensic medicine that uses microbes as clues,providing us information about lifestyle,circadian rhythms,geographic locations,postmortem intervals,cancers,and oral or systemic diseases.Oral cavity,as the place with the second largest number of microorganisms,can provide researchers with microbial information of each ecological niche,and assist in the prediction,diagnosis and monitoring of oral or systemic diseases.This paper reviews the composition of oral microbiome,the application in oral diseases,systemic diseases and forensic medicine,with the aim of providing some references for the development of forensic microbiology based on oral microbiome.
7.Improvement effect of acupuncture on locomotor function in Parkinson disease via regulating gut microbiota and inhibiting inflammatory factor release
Qiang WANG ; Yuan WANG ; Zhibin LIU ; Jie GUO ; Jie LI ; Yingqian ZHAO
Journal of Acupuncture and Tuina Science 2022;20(5):339-353
Objective: To observe the effect of acupuncture on the intestinal flora in Parkinson disease (PD) model mice and explore the mechanism of acupuncture in improving the locomotor function in PD. Methods: Thirty-two C57BL/6 mice were randomly divided into a control group, a 1-methyl-4-phenyl-1,2,3,6- tetrahydropyridine (MPTP) group, a MPTP + acupuncture group (MPTP+A), and a MPTP + madopar group (MPTP+M), with 8 mice in each group. Except for the control group, the other groups were intraperitoneally injected [25 mg/(kg·bw)] with MPTP to establish PD mouse models. After successful modeling, the MPTP group received no intervention, the MPTP+A received acupuncture at Tianshu (ST25), Guanyuan (CV4), and Zusanli (ST36), and the MPTP+M was given madopar [125 mg/(kg·bw)] by intragastric gavage. After consecutive 10-day interventions, the intestinal function and behaviors of the mice were detected. The 16S rRNA gene sequence was used to analyze the composition of fecal intestinal flora in each group. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and enzyme-linked immunosorbent assay were used to detect the expression levels of inflammatory cytokines in the brain and serum. The expression levels of tyrosine hydroxylase (TH) and α-synuclein in the substantia nigra (SN) were detected by immunohistochemical staining. Toll-like receptor (TLR) 2 and lipopolysaccharide receptor CD14 (CD14) in the SN were determined by RT-qPCR. Myeloid differentiation factor (MyD) 88, nuclear factor kappa-B (NF-κB) and Akt1 in the SN were detected by Western blotting. Results: After the intervention, compared with the control group, the intestinal motility, fecal water content, and the expression of TH in the SN were significantly decreased in the MPTP group (P<0.05), along with an increased α-synuclein expression (P<0.05). Additionally, the results of the fecal microflora test showed that the alpha diversity of the MPTP decreased, and the levels of inflammatory cytokines [tumor necrosis factor (TNF)-α, inducible nitric oxide synthase (iNOS), interleukin (IL)-1β, and IL-6] in the serum and SN, and the expression of NF-κB in the SN were significantly increased (P<0.05). Compared with the MPTP group, acupuncture intervention significantly enhanced the autonomous horizontal movement and coordination ability of PD mice (P<0.05); acupuncture and madopar interventions significantly reduced the levels of α-synuclein, inflammatory cytokines (TNF-α, iNOS, IL-1β, and IL-6) in the serum and SN, and the NF-κB expression in the SN, along with significantly increased alpha diversity richness index (P<0.05). In addition, the relative abundance of Bacteroides increased significantly in the MPTP+A (P<0.05), while the relative abundance of Firmicutes and Cyanobacteria decreased significantly (P<0.05). Conclusion: Acupuncture intervention can improve locomotor function, reduce α-synuclein aggregation and inflammatory factors expression, and increase the Akt signaling pathway in PD mice. In addition, acupuncture intervention can benignly regulate the intestinal flora of PD mice. Therefore, it suggests that acupuncture intervention can protect PD model mice probably by regulating intestinal flora and activating Akt signaling pathway.
8.Glutamine protects against oxidative stress injury through inhibiting the activation of PI3K/Akt signaling pathway in parkinsonian cell model.
Yingqian ZHAO ; Qiang WANG ; Yuan WANG ; Jie LI ; Gang LU ; Zhibin LIU
Environmental Health and Preventive Medicine 2019;24(1):4-4
BACKGROUND:
Parkinson's disease is a neurodegenerative disorder, and recent studies suggested that oxidative stress contributes to the degeneration of dopamine cell in Parkinson's disease. Glutamine also has a positive role in reducing oxidative stress damage. In this study, we hypothesized that glutamine offers protection against oxidative stress injury in 1-methyl-4-phenylpyridinium (MPP)-induced Parkinson's disease cell model.
METHODS:
MPP was used to induce PD models in PC12 cells and classified into control, M0 (MPP), G0 (glutamine), and M0+G0 groups. CCK-8 and AO/EB staining assays were used to examine cell proliferation and apoptosis, respectively. Western blotting was applied to examine the protein expression of PI3K, P-Akt, Akt, P-mTOR, and mTOR.
RESULTS:
We showed that glutamine suppressed cytotoxicity induced by MPP in PC12 cells. MPP decreased the superoxide dismutase and glutathione peroxidase activity and increased the malondialdehyde content, which were restored by glutamine. Moreover, MPP increased the expression of PI3K, P-Akt, Akt, P-mTOR, and mTOR, which were inhibited by glutamine. And the antioxidant capacity of glutamine on PC12 cells could be improved by LY294002 and inhibited by IGF-1.
CONCLUSION
These results suggest that glutamine strengthens the antioxidant capacity in PC12 cells induced by MPP through inhibiting the activation of the PI3K/Akt signaling pathway. The effects of glutamine should be investigated and the protective mechanism of glutamine in PD must be explored in future studies.
1-Methyl-4-phenylpyridinium
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administration & dosage
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Analysis of Variance
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Animals
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Cell Culture Techniques
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Disease Models, Animal
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Glutamine
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pharmacology
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Oxidative Stress
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drug effects
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Parkinson Disease
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Phosphatidylinositol 3-Kinases
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metabolism
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Protective Agents
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pharmacology
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Proto-Oncogene Proteins c-akt
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metabolism
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Rats
9.PCR detection of the Nocardia gene SecA1 using molecular beacon probe
Yanyan WANG ; Maoning XIA ; Chunyan MING ; Jing HUANG ; Taohua LIU ; Bing ZHOU ; Yingqian KANG
Chinese Journal of Zoonoses 2017;33(6):508-512
Molecular beacon probe was designed based on a specific DNA sequence of Nocardia to PCR detection of thisbacterium.The strains of Nocardia、Gordina and Rhodococcus were inoculated in Brain Heart Infusion Agar medium separately,then the growth condition was observed,DNA was extracted as a template;the molecular beacon probe was designed based on the partial secA 1 gene sequences of Nocardia strains,and the probe was added into the reaction system of real time fluorescence quantitative PCR (RT-PCR),and the fluorescence signal was tested at the end of PCR.Showed that the amplified secA1 gene of Nocardia could produce positive fluorescence signal in RT-PCR,but those of Gordonia and Rhodococcus with control groups showed negative results because of no fluorescence signal.In conclusion as a housekeeping gene,secA1 is an ideal target molecule to identify the actinomycetes strains on the species level in the systematic evolution research,and the technique of fluorescence molecular beacon probe is accurate,rapid and sensitive for detecting the Nocardia strains with secA1 gene.
10.Genomic sequencing analysis of Cryptococcus neoformans var grubii strains of two genotypes with dif-ferent virulence and selection of virulence-associated genes
Taohua LIU ; Yanyan WANG ; Yuru CHEN ; Liang ZHAO ; Qian LYU ; Lili MOU ; Yingqian KANG
Chinese Journal of Microbiology and Immunology 2016;36(2):103-109
Objective To analyze the genomic sequences of Cryptococcus neoformans var grubii strains of two genotypes with different virulence and to screen out the virulence-associated genes. Methods A clinical strain (IFM56800) with the strongest virulence and an environmental strain (IFM56731) with the weakest virulence were screened out for whole genome sequencing analysis. The results of sequencing analy-sis were comprehensively analyzed by using the method of comparative genomics. Genetic variations were ex-tensively screened by using the strategies of non-synonymous single nucleotide polymorphisms ( nsSNPs), nonsense SNPs and the insertions or deletions ( InDels) causing frameshift mutations. The filtered genes were sequenced in 20 experimental strains. The whole RNAs were extracted and then the full-length cDNAs were sequenced by using the rapid amplification of 5′ and 3′ cDNA ends (RACE) method. Results By whole genome sequencing, valid data with high coverage (127 times and 111 times) was obtained in both the environmental strain IFM56731 and the clinical strain IFM56800. The data of InDels and SNPs were statisti-cally analyzed, respectively. Six genes were chosen for further analysis based on the strategies of nonsense SNPs and the InDels causing frameshift mutations. The six genes were amplified and sequenced in all of the experimental strains, three of which were further analyzed with cDNA sequencing. Ultimately, the location and structure of CNAG_01032 gene were determined. The predicted nonsense mutation locus was verified to present in the actual mRNA. Conclusion The strategies of nonsense SNPs and the InDels causing frame-shift mutations showed high-efficiency in screening potential virulence-associated genes. The CNAG_01032 gene was screened out as a novel virulence-associated gene.

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