1.Differential Analysis of Oral Microbiota in db/db Mouse Model of Type 2 Diabetes Utilizing 16S rRNA Sequencing
Qianjia PAN ; Junyi GE ; Nan HU ; Fei HUA ; Min GU
Laboratory Animal and Comparative Medicine 2025;45(2):147-157
Objective To investigate the changes in oral microbiota of db/db mice and provide an experimental basis for exploring the relationship between type 2 diabetes mellitus and oral microecology.Methods Eight 10-week-old male db/db mice were designated as the diabetes experimental group(db/db group),while eight 10-week-old male db/m mice were assigned as the normal control group(db/m group).After a 5-day adaptive feeding period,tail venous blood samples were collected on the 6th and 37th days,and fasting blood glucose(FBG)levels and oral glucose tolerance test(OGTT)were performed for both groups to verify the reliability of the diabetes model.On the 15th day of feeding with the same diet,oral microbiota samples were collected from the buccal mucosa,dorsal and ventral tongue surfaces,oral floor mucosa,hard palate mucosa,and the gingival areas of both the upper and lower jaws of the two groups.Genomic DNA from the oral microbiota was extracted,and the V3-V4 regions of the 16S ribosomal RNA(16S rRNA)gene were amplified using a GeneAmp 9700 thermocycler.The composition of the oral microbiota was evaluated through double-labelled amplification and sequencing on the Illumina MiSeq platform,followed by bioinformatics analysis using QI I ME software(version 1.6.0).Results The FBG levels and OGTT results on the 6th and 37th days after the start of the experiment indicated that db/db mice exhibited more pronounced symptoms of type 2 diabetes compared to db/m mice.Alpha diversity(αdiversity)analysis showed no significant difference in the diversity of oral microbiota between the two groups(P>0.05);however,there was a significant difference in richness(P<0.05).Principal coordinate analysis(PCoA)revealed differences in the oral microbiota composition between the db/db group and db/m group(P<0.05).Species composition analysis and LEfSe analysis demonstrated that the relative abundance of oral microbiota in db/db group mice,predominantly composed of p_Proteobacteria,increased significantly at the phylum level(P<0.05).At the genus level,the relative abundances of g_Proteus and g_Enterococcus showed a significant increase(P<0.001).Conclusion The composition and diversity of oral microbiota in db/db mice with type 2 diabetes mellitus significantly differed from those without the disease.
2.Differential Analysis of Oral Microbiota in db/db Mouse Model of Type 2 Diabetes Utilizing 16S rRNA Sequencing
Qianjia PAN ; Junyi GE ; Nan HU ; Fei HUA ; Min GU
Laboratory Animal and Comparative Medicine 2025;45(2):147-157
Objective To investigate the changes in oral microbiota of db/db mice and provide an experimental basis for exploring the relationship between type 2 diabetes mellitus and oral microecology.Methods Eight 10-week-old male db/db mice were designated as the diabetes experimental group(db/db group),while eight 10-week-old male db/m mice were assigned as the normal control group(db/m group).After a 5-day adaptive feeding period,tail venous blood samples were collected on the 6th and 37th days,and fasting blood glucose(FBG)levels and oral glucose tolerance test(OGTT)were performed for both groups to verify the reliability of the diabetes model.On the 15th day of feeding with the same diet,oral microbiota samples were collected from the buccal mucosa,dorsal and ventral tongue surfaces,oral floor mucosa,hard palate mucosa,and the gingival areas of both the upper and lower jaws of the two groups.Genomic DNA from the oral microbiota was extracted,and the V3-V4 regions of the 16S ribosomal RNA(16S rRNA)gene were amplified using a GeneAmp 9700 thermocycler.The composition of the oral microbiota was evaluated through double-labelled amplification and sequencing on the Illumina MiSeq platform,followed by bioinformatics analysis using QI I ME software(version 1.6.0).Results The FBG levels and OGTT results on the 6th and 37th days after the start of the experiment indicated that db/db mice exhibited more pronounced symptoms of type 2 diabetes compared to db/m mice.Alpha diversity(αdiversity)analysis showed no significant difference in the diversity of oral microbiota between the two groups(P>0.05);however,there was a significant difference in richness(P<0.05).Principal coordinate analysis(PCoA)revealed differences in the oral microbiota composition between the db/db group and db/m group(P<0.05).Species composition analysis and LEfSe analysis demonstrated that the relative abundance of oral microbiota in db/db group mice,predominantly composed of p_Proteobacteria,increased significantly at the phylum level(P<0.05).At the genus level,the relative abundances of g_Proteus and g_Enterococcus showed a significant increase(P<0.001).Conclusion The composition and diversity of oral microbiota in db/db mice with type 2 diabetes mellitus significantly differed from those without the disease.
3.Application of deep learning to identify recurrent laryngeal nerve in endoscopic thyroidectomy via breast approach
Surong HUA ; Zhihong WANG ; Jiayi LI ; Junyi GAO ; Jing WANG ; Guanglin HE ; Palashate YEERKENBIEKE ; Xianlin HAN ; Ge CHEN ; Quan LIAO
Chinese Journal of Endocrine Surgery 2022;16(3):287-292
Objective:To explore whether deep learning could apply to recognize the recurrent laryngeal nerve (RLN) in videos of endoscopic thyroidectomy (ETE) via breast approach.Methods:Videos of ETE via breast approach in Peking Union Medical College Hospital from Feb. 2020 to Aug. 2021 were collected. Videos containing RLN were selected, and the outline of RLN was marked by two thyroid surgeons. Then data were divided into a training set and a test set in a ratio of 5:1 and classified into the high and low difficulty group according to a senior thyroid surgeon’s opinion. Those pictures were input to D-LinkNet model. Precision, sensitivity and mean dice index was calculated.Results:A total of 46 videos including 153, 520 frames of pictures were included in this study. 131,039 frames of 39 videos were in the training set and 22,481 frames of 7 videos were in the test set. When the intersection over union threshold was 0.1, the sensitivity and precision was 92.9%/72.8% and 47.6%/54.9% in high and low recognition group, respectively. When the intersection over union threshold was 0.5, the sensitivity and precision turned to 85.8%/67.2% and 37.6%/43.5% in high and low difficulty group, respectively. Mean Dice index was 0.781 and 0.663 in high and low difficulty group, respectively.Conclusions:RLN recognition based on deep learning is feasible and has potential application value in ETE, which may help surgeons reduce the risk of accidental injury of RLN and improve the safety of thyroidectomy.
4.Application of Endoscopic Parathyroidectomy in the Treatment of Primary Hyperparathyroidism
Surong HUA ; Zhihong WANG ; Junyi GAO ; Mengyi WANG ; Qiaofei LIU ; Wenjing LIU ; Guannan GE ; Yingxin WEI ; Ya HU ; Quan LIAO
Chinese Journal of Endocrine Surgery 2022;16(4):391-395
Objective:To summarize the experience and the clinical data of patients with primary hyperparathyroidism undergoing endoscopic parathyroidectomy.Methods:A total of 24 patients who underwent endoscopic parathyroidectomy for primary hyperparathyroidism in Peking Union Medical College Hospital during Feb. 2021 to May. 2022 were concluded in this study (20 cases of parathyroidectomy via axillary approach and 4 cases of parathyroidectomy via thoracic and breast approach) . The operation time, postoperative drainage, length of stay, level of parathyroid hormone and serum calcium of those patients were collected. Postoperative complications and recurrence of hyperparathyroidism were also observed.Results:The postoperative levels of serum parathyroid hormone and serum calcium were significantly reduced (over 50%) compared with preoperative level ( P<0.05) . The average operation time was (96±22) min (64-157 min) . The mean postoperative drainage volume was (47±16) ml on day 1, (46±11) ml on day 2, and (30±9) ml on day 3, respectively. The average length of postoperative hospital stay was (2.8±1.1) days (2-6 days) . In one case of parathyroidectomy via axillary approach, the operation was converted to open surgery because of the low position of lesion. Other cases completed endoscopic surgery and obtained satisfactory cosmetic results. There were no postoperative complications such as bleeding, permanent hoarseness, coughing while drinking water, or surgical site infection. The mean follow-up time was (7.4±4.2) months (1-16 months) . There was no obvious discomfort and no recurrence during follow-up. Conclusion:Endoscopic parathyroidectomy is safe and effective in the treatment of primary hyperparathyroidism, which can be used as a surgical option for patients with cosmetic requirements.
5.Exploration of deep learning to identify recurrent laryngeal nerve in endoscopic thyroidectomy via unilateral axillary approach
Surong HUA ; Zhihong WANG ; Junyi GAO ; Jing WANG ; Guanglin HE ; Xianlin HAN ; Ge CHEN ; Quan LIAO
Chinese Journal of Endocrine Surgery 2022;16(1):5-11
Objective:To explore whether deep learning could apply to recognize the recurrent laryngeal nerve in the video of unilateral axillary approach endoscopic thyroidectomy.Methods:Videos of endoscopic thyroidectomy via unilateral axillary approach in Peking Union Medical College Hospital from Jul. 1st, 2020 to May. 1st, 2021 were collected. Videos containing the recurrent laryngeal nerve were selected, and the outline of recurrent laryngeal nerve were marked by two senior thyroid surgeons and staffs. Data were divided into training set and test set in a ratio of 5:1, and classified into high, medium and low recognition group according to difficulty of recognizing the outline of the nerve. The neuron network was based on PSPNet combined with Resnet50. All data were analyzed by R (ver. 4.0.2) .Results:A total of 38 videos including 35,501 frames of pictures were included in this study. 29, 704 frames of 32 videos were in our training set and 5797 frames of 6 videos were in the test set. When the intersection over union (IOU) threshold is 0.1, the sensitivity and precision is 100.0%/92.1%, 95.8%/80.2% and 81.0%/80.6% in high, medium and low recognition group respectively. When the IOU threshold is 0.5, the sensitivity and precision is 92.6%/85.3%, 71.7%/60.5% and 38.1%/37.9% in high, medium and low recognition group respectively, indicating that neuron network could located the outline of recurrent laryngeal nerve in high and medium recognition group. False negatives were often due to small targets and unclear boundaries.Conclusion:Recurrent laryngeal nerve recognition based on deep learning is feasible and has potential application value in endoscopic thyroidectomy, which may help surgeons reduce the risk of accidental injury of recurrent laryngeal nerve and improve the safety of thyroidectomy.
6.Beneficial Effects of Celastrol on Immune Balance by Modulating Gut Microbiota in Experimental Ulcerative Colitis Mice
Li MINGYUE ; Guo WEINA ; Dong YALAN ; Wang WENZHU ; Tian CHUNXIA ; Zhang ZILI ; Yu TING ; Zhou HAIFENG ; Gui YANG ; Xue KAMING ; Li JUNYI ; Jiang FENG ; Sarapultsev ALEXEY ; Wang HUAFANG ; Zhang GE ; Luo SHANSHAN ; Fan HENG ; Hu DESHENG
Genomics, Proteomics & Bioinformatics 2022;20(2):288-303
Ulcerative colitis(UC)is a chronic inflammatory bowel disease caused by many factors including colonic inflammation and microbiota dysbiosis.Previous studies have indicated that celastrol(CSR)has strong anti-inflammatory and immune-inhibitory effects.Here,we investigated the effects of CSR on colonic inflammation and mucosal immunity in an experimental colitis model,and addressed the mechanism by which CSR exerts the protective effects.We characterized the ther-apeutic effects and the potential mechanism of CSR on treating UC using histological staining,intestinal permeability assay,cytokine assay,flow cytometry,fecal microbiota transplantation(FMT),16S rRNA sequencing,untargeted metabolomics,and cell differentiation.CSR administra-tion significantly ameliorated the dextran sodium sulfate(DSS)-induced colitis in mice,which was evidenced by the recovered body weight and colon length as well as the decreased disease activity index(DAI)score and intestinal permeability.Meanwhile,CSR down-regulated the production of pro-inflammatory cytokines and up-regulated the amount of anti-inflammatory mediators at both mRNA and protein levels,and improved the balances of Treg/Thl and Treg/Th1 7 to maintain the colonic immune homeostasis.Notably,all the therapeutic effects were exerted in a gut microbiota-dependent manner.Furthermore,CSR treatment increased the gut microbiota diversity and changed the compositions of the gut microbiota and metabolites,which is probably associated with the gut microbiota-mediated protective effects.In conclusion,this study provides the strong evidence that CSR may be a promising therapeutic drug for UC.
7. Proteomic analysis of serum exosomes derived from Crouzon syndrome patients carrying a mutant FGFR2C342Y
Min GE ; Yingzhi WU ; Ronghu KE ; Tianyi CAI ; Junyi YANG ; Xiongzheng MU
Chinese Journal of Plastic Surgery 2018;34(4):299-304
Objective:
Reveal the global expression profile of serum exosomal proteins of Crouzon syndrome patients.
Methods:
We isolated microvesicles from serum of Crouzon children with a C342Y mutation in FGFR2 by ultracentrifugation, which were further characterized by electron microscopy and immunoblotting. The protein profiling in normal subjects and Crouzon patients was systematically compared by iTRAQ proteomic analysis.
Results:
The result demonstrated that microvesicles were between 30—100 nm in diameter, round shape with cup-like concavity and expressed exosomal marker tumor susceptibility gene (TSG) 101 and flotillin (Flot) 1. We identified a total number of 62 proteins, among which 22 proteins overlap with ExoCarta database and were different between the Crouzon patient and the normal subject. The Ingenuity Pathway Analysis showed that the functions of these proteins are mostly involved in Developmental Disorder, Hereditary Disorder, Skeletal and Muscular Disorders, which are all related to the clinical manifestations of Crouzon syndrome. In addition, the proteins were focused on the network of "Organismal Injury and Abnormalities, Hematological System Development and Function, Cell-To-Cell Signaling and Interaction" . The central protein FN1 was presented as the key protein in the network.
Conclusions
Our data demonstrated that serum exosomes harbor informative proteins and FN1 was selected as a potential candidate for its role in promoting osteoblast adhesion, proliferation and mineralization.
8.An analysis of the status quo of nurses' career planning in planning two-child nurses and the countermeasures
Xiaoling FENG ; Junyi CHEN ; Jun GE
Modern Clinical Nursing 2017;16(7):62-66
Objective To explore the status quo of the career planning of the second child nursery under the two-child fertility policy, and to provide reference for carrying out the targeted management intervention. Method From April to May of 2017, 592 nurses of the second child were surveyed using the general data questionnaire and the career planning scale. Results The nurses' career planning score (34.81 ± 3.59) was different, and the career planning of the two nurses with different age, job title, title, establishment, department and income and fertility was different and statistically significant. Nurses with plan to a second child are with less desire to further study and promotion than those without plan to a second child (all P<0.001) promotion to open more than two children before the low policy (P <0.05). Conclusion Nurses' career planning for births is on a middle level. Nursing managers should pay attention to the management of nurses' career planning , create a career development environment , provide humanistic care , strengthen self-awareness and career self-development of nurses who have two births , so as to improve the level of their career planning to promote the nursing team development.
9.Simultaneous Determination of 5 Main Components in the Fruits and Root of Wild Acanthopanax senticosus by UPLC
Huimin YAO ; Yingli GUAN ; Junyi ZHU ; Yanjie GE ; Yuan HUANG ; Meng ZHANG
China Pharmacy 2016;27(12):1668-1671
OBJECTIVE:To establish a method for the 5 main components (original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside) in the fruits and roots of wild Acanthopanax senticosus. METHODS:UPLC was per-formed on the column of Waters ACQUITY UPLC HSS T3 with mobile phase of acetonitrile-0.3% phosphoric acid (gradient elu-tion)at a flow rate of 0.2 ml/min. Detection wavelength was 300 nm,column temperature was 30 ℃,and injection volume was 10μl. RESULTS:The linear range was 24.56-184.2 μg/ml for syringin(r=0.9993),18.454-138.405 μg/ml for chlorogenic acid(r=0.9993),8.416-63.12 μg/ml for eleutheroside E (r=0.9997),3.286-24.645 μg/ml for isofraxidin (r=0.9993) and 2.522-18.915μg/ml for quercetin-3-rhamnoside(r=0.9998);RSDs of precision,stability and reproducibility tests were lower than 1%;recover-ies were 99.14%-100.50%(RSD=0.48%,n=6)for syringing in the fruits of A. senticosus、99.03%-100.45%(RSD=0.50%,n=6) for chlorogenic acid in the fruits of A. senticosus、99.22%-100.44%(RSD=0.44%,n=6)for eleutheroside E in the fruits of A. sen-ticosus、99.80%-100.80%(RDS=0.44%,n=6)for isofraxidin in the fruits of A. senticosus、99.76%-101.10%(RSD=0.51%,n=6) for quercetin-3-rhamnoside in the fruits of A. senticosus;99.21%-101.20%(RSD=0.73%,n=6)for syringing in the root of A. senti-cosus、99.81%-101.20%(RSD=0.52%,n=6)for chlorogenic acid in the root of A. senticosus、100.00%-101.50%(RSD=0.62%, n=6)for eleutheroside E in the root of A. senticosus、99.22%-100.40%(RSD=0.47%,n=6)for isofraxidin in the root of A. senti-cosus. CONCLUSIONS:The method is simple and stable with good reproducibility,and can be used for the simultaneous determi-nation of original syringin,chlorogenic acid,eleutheroside E,isofraxidin and quercetin-3-rhamnoside in the fruits and root of wild A. senticosus.
10.Clinical Study of the Effect of Electroacupuncture on Rehabilitation and TCD in Convalescent Stroke Patients
Runjia ZHU ; Mingshu XU ; Linbao GE ; Chunyan CHEN ; Xiaolan SU ; Ling ZHANG ; Yanli ZHOU ; Junyi WU
Shanghai Journal of Acupuncture and Moxibustion 2015;(5):389-392
Objective To investigate the clinical therapeutic effect of electroacupuncture on ischemic stroke by recording the therapeutic effect score and performing transcranial Doppler (TCD) detection in electroacupuncture-treated convalescent stroke patients before and after treatment.Methods Sixty convalescent patients with ischemic stroke were randomly allocated to treatment and control groups, 30 cases each. The control group received routine medication and the treatment group, electroacupuncture in addition. In the two groups, pre- and post-treatment neurological defect scores were recorded and the clinical therapeutic effects were observed after four weeks of treatment. Pre- and post-treatment middle cerebral artery (MCA) pulsation indices (RI), resistance indices (RI) and ratios of systolic to end-diastolic (S/D) blood flow velocity detected by TCD were compared between the two groups. Results The total efficacy rate was 80.0% in the treatment group and 50.0% in the control group; there was a statistically significant difference between the two groups (P<0.01). There was a statistically significant pre-/post-treatment difference in the MESSS score in the two groups (P<0.01). There was a statistically significant post-treatment difference in the MESSS score between the treatment and control groups (P<0.05). There were statistically significant pre-/post-treatment differences in TCD indices (PI, RI and S/D) on the affected side in the two groups (P<0.05). There were statistically significant post-treatment differences in TCD indices on the affected side between the treatment and control groups (P<0.05).Conclusion Electro- acupuncture can promote rehabilitation in convalescent stroke patients.

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