Abstract:Objective：This stuly aims to explore strategies for effectively studying the interaction between membrane receptors and extracellular ligands using fluorescence resonance energy transfer(FRET). Methods：Based on the model of interaction between LRP6 and WNT3A established by PDB and Zdock database，LRP6 was truncated without affecting the interaction between LRP6 membrane receptor and WNT3A；the signal peptides of LRP6 and WNT3A were reconstituted at the amino terminus of the fluorescent tag to avoid loss of the fluorescent tag due to signal peptide cleavage；flexible linker peptides was used to connect signal peptides，fluorescent tags， and mature proteins to attenuate the effects of fluorescent tags on subcellular localization and function of membrane receptors. Results：The above improvement strategy significantly enhanced the membrane localization signal of LRP6 fusion protein. The interaction between LRP6 and WNT3A in the cell membrane region could be clearly detected by FRET technology，and the average FRET efficiency reached 26.51%. Conclusion：This study provides new ideas and strategies for effectively using FRET technology to study the interaction between membrane receptors and extracellular ligands in living cells.

Abstract:Objective：To explore the neuroprotective effects and the underlying mechanisms of shikonin(SK)after traumatic brain injury(TBI). Methods：Forty C57BL/6 mice were randomly assigned to 4 groups as follows：sham operation group(Sham group)，TBI group，TBI + 1 mg/kg shikonin group(TBI+SK 1 mg/kg)，TBI + 5 mg/kg shikonin group(TBI+SK 5 mg/kg). The modified neurological severity scores(mNSS)and the apoptosis of neurons after TBI(Neuron/TUNEL)was observed，the integrity of the blood brain barrier (BBB)was detected by Evans blue staining. Moreover，we used Western blot and RT⁃PCR to determine the expression of NLRP3/ASC/ IL⁃1β/Caspase1 and detected the changes in the levels of oxidative stress markers including ROS，LPO，MDA and antioxidant enzymes which includes SOD and GPx in the edema zone around the cortical injury. The Neuro⁃2a cell line and BV2 cell line were cultured in vitro，stimulated by LPS to establish the inflammatory environment. We used Western blot to observe the inflammatory response of BV2cells and observed that the changes of neuronal apoptosis by flow cytometry. Results：Shikonin treatment improved mNSS after TBI and exerted neuroprotective effects. Shikonin could inhibit the level of ROS，LPO，MDA，and promoted the increase of SOD，GPx， alleviating the oxidative damage after TBI. Moreover，shikonin significantly inhibited the inflammatory activation of NF ⁃ κB/NLRP3 after TBI，and inhibited the activation of microglia and astrocytes in the penumbra around the injury. Conclusion：Shikonin may play a neuroprotective effect after TBI via inhibiting the activation of NF⁃κB/NLRP3 and alleviating oxidative stress.

Abstract:Objective：A novel multidrug resistant(MDR)Escherichia coli(E.coli)strain J2_5 was isolated from a patient’s mid ⁃ stream urine sample. According to the genome sequence of J2_5，we analyzed its evolutionary classification，drug resistance genes and virulence genes，providing theoretical basis for clinical diagnosis and drug selection for drug resistant bacterial infection. Methods： Genome DNA of J2_5 was isolated and sequenced by high ⁃ throughput sequencing technology，after analyzed the high ⁃ throughput sequencing data and compared it with known databases by a variety of bioinformatics tools，a high integrity genome of J2_5 was obtained. Results：The total length of the whole genome of J2_5 was 4.7Mb and GC content was 50.78%. Phylogenetic tree analysis showed that J2_ 5’genome was closely related to that of wild type E. coli strain MG1655. But Multilocus sequence typing(MLST)analysis revealed ST type of J2_5 was ST2491，which was different from that of MG1655(ST10). Furthermore，CARD database analysis showed that the genome of J2_5 encodes 65 drug resistance genes，which make the strain obtain drug⁃resistant ability through four different mechanisms (antibiotic efflux pump，antibiotic inactivation，antibiotic target change，reduced permeability to antibiotics). VFDB database analysis showed that there were 40 virulence genes in 7 categories，encoding adhesive proteins，invasion proteins，autonomic transporters and other toxin proteins. After analyzing the whole genome of J2_5 through the Prophage Hunter online tool，we found that there were three prophages in the genome of J2_5，and a virulence gene was carried by one of the prophages. Conclusion：The whole genome sequencing results showed that J2_5 carries some virulence genes and drug resistance genes，which makes the strain obtain strong pathogenicity and resistance ability to a variety of antibiotics.

Abstract:Objective：To determine the nuclear localization signal of vitamin D receptor(VDR)and verify its function. Methods： The nuclear localization signal of VDR was predicted by software analysis and literature investigation. A series of point mutants of VDR were constructed by a point mutation kit. The distribution of VDR in the cytoplasm and nucleus was obsened by Western blot and immunofluoresceme to determine the effect of mutation points in the entry of VDR into the nucleus. Finally，quantitatve real⁃time PCR (qPCR)was used to detect the expression of genes downstream of VDR to determine the activity of VDR nuclear localization mutants. Results：The nuclear localization signal sequence of VDR was determined to be49RRSMKRKALFLT61. The expression of a series of NLS (nuclear localization sequence)point mutants of VDR was mainly distributed in the cytoplasm. Vitamin D could neither induce these point mutants into the nucleus nor promote the expression of downstream genes of vitamin D. Conclusion：VDR nuclear localization signal can effectively control the entry of VDR into the nucleus，play the role of transcription factors，and then affect the expression of downstream genes，which provides a new platform and idea for exploring the treatment of vitamin D and VDR related diseases。

Abstract:Objective：To explore the effect and mechanism of TIR/BB⁃loop mimetic AS⁃1 on renal ischemia/reperfusion injury in mice. Methods：Male C57BL/6 mice were randomly and equally divided into four groups：sham operation group(Sham group)，renal ischemia/reperfusion injury group(RIR group)，AS ⁃ 1+ renal ischemia/reperfusion injury group(AS ⁃ 1+RIR group)，vehilce + renal ischemia/reperfusion injury group(vehilce + RIR group). The renal ischemia/reperfusion injury model was constructed by clipping bilateral renal veins for 45 min and reperfusion for 24 h. The AS⁃1+RIR group and vehilce+RIR group were intraperitoneally injected with 50 mg/kg AS ⁃ 1 and vehicle 30 minutes before operation. After 24 h reperfusion，renal functional parameters，serum mediator concentrations，markers of apoptosis indexes and expression of phosphorylated NF⁃κB p65 were determined. Results：Serum creatinine (Scr)、blood ureanitrogen(BUN)、TNF⁃α、IL⁃6、Cleaved caspase3、Bax and p⁃NF⁃κB p65 were significantly increased after renal IR compared with the sham group(P＜0.01). The expression of Bcl⁃2 and the ratio of Bcl⁃2/Bax were significantly decreased(P ＜ 0.01). After treatment with AS⁃1，the levels of Scr、BUN、TNF⁃α、IL⁃6、cleaved caspase 3、Bax and P⁃p65 were significantly decreased(P ＜ 0.01)，the expression of Bcl ⁃2 was increased(P ＜ 0.01)，and the ratio of Bcl ⁃2/Bax was also increased. Conclusion：TIR/BB loop analogue AS ⁃ 1 has protective effect on renal ischemia ⁃ reperfusion injury in mice. The mechanism may be related to its anti ⁃ inflammatory and anti apoptotic effects by inhibiting the activation of NF ⁃ κ B signaling pathway.

Abstract:Objective：To investigate the potential influence of thrombospondin⁃4(TSP⁃4)on the proliferation，apoptosis，oxidative stress and inflammatory response in podocytes stimulated by high glucose(HG)，thus revealing the function of TSP ⁃ 4 in the development of diabetic kidney disease(DKD). Methods：① Relative levels of TSP ⁃ 4 and signal transducer and activator of transcription3(STAT3)in HG⁃induced podocytes were examined by real⁃time quantitative PCR(qRT⁃PCR)and Western blot.②After intervention of TSP⁃4 in HG⁃induced podocytes，proliferative ability was examined by cell counting kit⁃8(CCK⁃8)and 5⁃Ethynyl⁃2’⁃ deoxyuridine(EdU)assay. The apoptosis and cell cycle distribution of podocytes were determined by flow cytometry. Relative contents of reactive oxygen species(ROS)，malondialdehyde(MDA)，superoxide dismutase(SOD)and catalase(CAT)were detected by commercial kits. In addition，the expression levels of inflammatory factors interleukin(IL)⁃1β，tumor necrosis factor(TNF)⁃α and IL⁃6 were measured by enzyme ⁃linked immunosorbent assay. ③The interaction between the transcription factor STAT3 and the promoter region of TSP ⁃ 4 was validated by chromatin immunoprecipitation(ChIP)and further confirmed by dual ⁃ luciferase reporter assay. Results：①TSP⁃4 and p⁃STAT3 were timE-dependently upregulated in HG⁃induced podocytes(P ＜ 0.05). ②Knockdown of TSP⁃4 could reverse the inhibited proliferation，apoptosis，oxidative stress and inflammatory response in HG⁃induced podocytes(P ＜ 0.05). ③ STAT3 could bind the promoter region of TSP⁃4，thus inducing the transcription，and potitively regulating the TSP⁃4 promoter activity.Conclusion：Knockdown of TSP ⁃ 4 alleviates HG ⁃induced apoptosis，oxidative stress and inflammatory response in podocytes，thus showing a protective effect on podocytes. Transcription factor STAT3 has positive function to the promoter activity of TSP⁃4.

Abstract:Objective：To investigate the proteomics of primary neurons exposed to methamphetamine(METH)and the corresponding cellular functions，unraveling the potential neurotoxicity of METH exposure. Methods：The primary cultured neurons were treated with METH(0，900 μmol/L)and then hydrolyzed by protease. LC⁃MS/MS was used to detect and quantify the proteins， then the database of Uniprot RattusNorvegicus_ 36080_ 20180123 was used to identify the target protein，gene ontology(GO)was used to annotate the protein function，and Kyoto Encyclopedia of Genes and Genomes(KEGG)database was used to classify the target protein sequence by KO analysis to obtain the target protein associated signal pathways. Based on the above pathways，Western blot was used to verify the synaptic damage induced by METH，and immunofluorescence was performed to examine the expression and spatial localization of synapses. Results：The total number of proteins identified was 6619. Compared with the control group，the protein with expression difference more than 1.2 times and P ＜ 0.05 was classified as differential protein. Through screening，51 different proteins were screened，of which 40 proteins were up ⁃ regulated while 11 proteins were down ⁃ regulated. For protein expression，METH exposure significantly decreased the expression of post synaptic density protein 95(PSD95)and Drebrin，while presynaptic marker synapsin 1 was significantly up ⁃ regulated；immunofluorescence results showed that Drebrin expression was significantly decreased accompanied by reduced co ⁃ localization with F ⁃ actin，meanwhile，the spatial localization of PSD95 and synapsin 1 was significantly decreased. Conclusion：METH exposure causes abnormal expression of numerous proteins in primary neurons as well as the disorderof a spectrum of cellular functions，which finalizes the destruction of synaptic structure and neuronal damage.

Abstract:Objective：To investigate the role of differential genes and lncRNA RAD51⁃AS1 in primary osteoporosis(OP)，which provides new insights for the diagnosis and treatment of primary osteoporosis. Methods：The GSE35956 microarray data from the GEO database was used for analysis. Differentially expressed genes(DEG)were obtained by using the DEseq2，and the ClusterProfiler was used to find the enrichment of DEGs. The microarrays were then reannotated to discover the differential long ⁃ non coding RNA (lncRNA)in OP. The expression of RAD51⁃AS1 in hBMSC was verified by qRT⁃PCR. Finally，the effects of RAD51⁃AS1 on hBMSC proliferation and osteogenic differentiation were examined by MTT，colony formation，and ALP staining. Results：A total of 1 440 DEG was obtained about mRNAs in this study，whose functions were mainly enriched in processes such as receptor ⁃ ligand activity and monocyte differentiation. A total of 105 DEG regarding lncRNAs were also acquired. We found that RAD51 ⁃ AS1 expression was significantly downregulated in osteoporosis. The proliferation and ALP activity of hBMSC was reduced after silencing RAD51⁃AS1. Conclusions：The PI3K ⁃Akt signaling pathway and monocyte differentiation plays important roles in osteoporosis by bioinformatics. LncRNA RAD51 ⁃ AS1 regulates the proliferation and differentiation of bone marrow mesenchymal stem cells to ameliorate the progression of osteoporosisby cytological verification.

Abstract:Objective：This srudy aims to establish a patient ⁃derived xenograft(PDX)model of NOD background immunodeficient mice for human cancer preclinical and biological research. Methods：NOD ⁃ PrkdcscidIl2rgnull mice were obtained by CRISPR/cas9 technology，and homozygous offspring with stable inheritance were obtained through normal breeding. The proportion of immune cells in peripheral blood of mice was monitored by flow cytometry. Fresh tumor tissues were transplanted and inoculated. The tumor growth status and the morphological characteristics of passaged PDX tumors were observed and recorded. Results：There was no expression of mature T，B and NK cells in peripheral blood of NYG mice. There was almost no rejection to human cells and tissues，and there was no obvious abnormality in histopathological phenotype and immune system. Conclusion：NYG mice and immunodeficient mice were successfully constructed，which provided a new PDX animal model for the basic theory and application of oncology.

Abstract:Objective：To explore the potential of the circular RNA hsa_circ_0005579 as a new biomarker for the development of pre ⁃eclampsia，and to observe its expression in the placenta of patients with prE-eclampsia and its effect on the proliferation，migration and invasion of trophoblast cells. Methods：The expression of hsa_circ_0005579 was detected in placental tissues obtained from 20 women with pre ⁃ eclampsia and 20 healthy pregnancy controls using real ⁃time quantitative polymerase chain reaction(qRT ⁃ PCR). Clinical information of participants was collected. Ribonuclease R(RNase R)digestion analysis，agarose gel electrophoresis and Sanger sequencing were used to verify hsa_circ_0005579. HTR⁃8/SVneo cells were transfected with hsa_circ_0005179 siRNA，cell counting kit⁃8(CCK⁃8)assay，colony formation assay，EdU test were performed to detect the effect of hsa_circ_0005579 on the proliferation of trophoblast cells. Transwell assay and wound healing assay were used to detect the effect of hsa_circ_0005579 on the migration and invasion of trophoblast cells. Meanwhile，Western blot analysis was used to detect the protein level of migration markers. Results： Hsa_circ_0005579 had a significant high expression in the placenta of PE patients. In addition，compared with the si ⁃NC group，the cell viability was significantly increased，the number of clone formation grew，the cell proliferation rate increased，and the number of migration and invasion did rise，the cell migration rate was upregulated，and the protein levels of migration markers matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9)was enhanced in the si⁃circ_0005579 group. Conclusion：CircularRNA hsa_circ_0005579 is highly expressed in the placenta of patients with pre ⁃ eclampsia，and the proliferation，migration and invasion ability of HTR ⁃ 8/SVneo is enhanced after silencing hsa_circ_0005579. It is expected to provide new targets for early diagnosis and intervention of prE-eclampsia.

Abstract:Objective：To establish a CRISPR/Cas13a based method for detecting TP53 R248W mutant molecules. Methods：TP53 R248W mutant plasmid was constructed by Over⁃lap PCR using TP53 wild⁃type plasmid as template. The CRISPR/Cas13a method for the detection of TP53 R248W variant was initially established by optimizing the amplification product size，amplification technology， the length and concentration of crRNA. The sensitivity of CRISPR/Cas13a method was evaluated by TP53 R248W variants with different mutation rates and simulated plasma ctDNA. Results：The size of the amplified product detected by CRISPR/Cas13a was about 368 bp. The concentration of crRNA had influence upon the detection intensity of CRISPR/Cas13a. In the range of 0.05~0.25μmol/L， the higher the concentration of crRNA，the higher the detection intensity of CRISPR/Cas13a was detected. The sensitivity of CRISPR/ Cas13a in detecting TP53 R248W variants was 104 copies/μL，and the minimum mutation rate was 0.01% . Conclusion：The established CRISPR/Cas13a method has the advantages of rapid，simple，sensitive and specific，and provides a new technology for the detection of TP53 R248W variant in tissues and plasma.

Abstract:Objective：The study aims to provide new targets for the prevention and treatment of newly diagnosed T2DM with HUA by analyzing the clinical features and the gut microbiome.Methods：A total of 103 patients with newly diagnosed T2DM and 98 patients with T2DM with HUA were included. According to the admission conditions，agE- and gender⁃matched newly diagnosed T2DM patients with HUA(DMUA group) and newly diagnosed T2DM patients(T2DM group) were selected，and age ⁃ and gender ⁃ matched hyperuricemia patients(HUA group)and healthy volunteers(Control group)were included at the same time. The clinical data and biochemical test results were recorded. The data were analyzed by chi⁃square test，analysis of variance，Tukey⁃HSD Test and Wilcoxon rank sum test method. Stool samples of 4 groups were collected and sequenced by 16S rRNA high⁃throughput sequencing method and all the data were finally analyzed by bioinformatics. Results：①Comparison of clinical features：CHO，TG，AI were higher in DMUA than T2DM group，LDL in DMUA was only higher than the T2DM group with a history of more than 5 years. DMUA group were youngerand had a greater prevalence of fatty liver than T2DM group，but HbA1c in DMUA was lower than T2DM group. Compared with the T2DM group，HUA group and the Control group，the DMUA group had a significant decrease in high ⁃ density lipoprotein and a significant increase in triglycerides. And compared with the T2DM group and HUA group，the arteriosclerosis index of DMUA groups was significantly increased.②The gut microbiome analysis：The α diversity index of the DMUA group was significantly lower than that of the Control group. There was a significant difference in β diversity between the DMUA group，Control group and the HUA group. In the analysis of bacteria abundance，compared with the HUA group，f_Prevotellaceae and g_Megamonas were significantly increased in the DMUA group，while p_Bacteroidetes，c_Bacteroidia，o_Bacteroidales，f_Bacteroidaceae，g_Bacteroides，f_Tannerellaceae and g_Parabacteroides were significantly decreased. Compared with the Control group， c_clostridia， o_clostridiales， f_Peptostreptococcaceae and g_Romboutsia were significantly decreased in the DMUA group. Conclusion：Compared with long⁃course T2DM with HUA and newly diagnosed T2DM alone，the group with newly diagnosed T2DM with HUA is more likely to have abnormal lipid metabolism. The gut microbiome of the DMUA group is different from that of the T2DM group，HUA group or Control group，which may be related to the occurrence and development of T2DM and HUA.

Abstract:Objective：To study the differential proteomics in exosomes of peritoneal dialysis effluent(PDE)from patients with different dialysis ages. Methods：Ten stable peritoneal dialysis(PD)patients were selected and divided into newly enrolled patients group(NEPs group，n=5)and maintenance peritoneal dialysis patients group(MPDs group，n=5)according to the dialysis age. PDE was collected from patients left overnight. Exosomes were extracted by ultracentrifugation and identified by transmission electron microscopy(TEM)，nanoparticle tracking analysis(NTA)，and Western blotting. Label ⁃free quantitative proteomics technology was used to identify and screen the exosomes. Functional enrichment and signal pathway bioinformatics analysis were performed for significantly differential proteins in the exosomes. Results：The exosomes in PDE showed saucer ⁃like vesicles under TEM，and the diameters were between 30 and 150 nm measured by NTA. The specific markers of exosomes，CD63 and TSG101 were detected by Western blotting. A total of 499 proteins were detected by proteomics analysis. After significant difference screening，17 were up ⁃ regulated and 15 were down ⁃ regulated. Gene ontology(GO)analysis showed that these differentially expressed proteins were mainly distributed in chloride channel complexes，cell surface and nuclear matrix，playing a role mainly in binding and catalytic activities，and were involved in biological processes such as positive regulation of binding，oxidative stress response and peroxidase response. Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways were mainly enriched in immunE-related signaling pathways，including IL⁃17signaling pathway and Th17 cell differentiation signaling pathway，which play a key role in peritoneal injury. Conclusion：The differentially expressed proteins screened by label ⁃ free quantitative proteomics technology could be used as candidate markers of peritoneal injury in PD patients and provids molecular clues for revealing the peritoneal fibrosis mechanism of pathogenesis.

Abstract:Objective：This study aims to investigate the safety and efficacy of transjugular intrahepatic portosystemic stent shunt (TIPS)combined with percutaneous transhepatic variceal embolization(PTVE)in the treatment of esophagogastric variceal bleeding in liver cirrhosis caused by autoimmune liver diseases(AILD).Methods：From August 2015 to September 2021，the clinical data of 23 patients(2 males and 21 females)with esophagogastric variceal bleeding in liver cirrhosis caused by AILD were retrospectively analyzed. All patients received TIPS and PTVE treatment after conservative and/or endoscopic treatment failed to control bleeding. The immediate bleeding control rate，rebleeding rate，rebleeding⁃free survival，overall survival and complications were evaluated. Results： Patients showed a high immediate bleeding control rate(100%). The 1⁃，2⁃，and 3⁃year rebleeding rate were 9.6%，17.1%，and 17.1%， respectively. The median rebleeding⁃free survival time was 455 days(range：70⁃1 308 days). The 1⁃，2⁃，3⁃year survival rate was 100%， 91.7%，and 91.7%，respectively. The median overall survival time was 611 days(range：85 ⁃ 1 308 days). The severe short ⁃ term complications included acute cardiac insufficiency(1 case)and intra ⁃abdominal hemorrhage(1 case). Long ⁃term complications were hepatic encephalopathy(4 cases)，with West Haven grade Ⅲ in 1 case and West Haven Grade Ⅱ in other 3 cases. Conclusion：TIPS combined with PTVE may be safe and effective in the management of esophagogastric variceal bleedingin liver cirrhosis caused by AILD.

Abstract:Objective：To investigate the maternal and infant outcomes and clinical characteristics of uterine rupture. Methods：The clinical characteristics and maternal and infant outcomes of 70 patients with uterine rupture who delivered in Nanjing Medical University Obstetrics and Gynecology Hospital from 2009 to 2020 were retrospectively analyzed according to whether there was scar uterus or not and the time point when uterine rupture was found. Results：① The incidence of uterine rupture in this study was 0.054%. ② Clinical manifestations：23 cases(32.86%)had abdominal pain，13 cases(18.57%)had vaginal bleeding，9 cases(12.86%) had fetal heart abnormalities，7 cases(10%)had hemodynamic instability，11 cases(15.71%)had bloody amniotic fluid，pathological contraction of loops and other manifestations，and 29 cases(41.43%)had no clinical manifestations. ③ Maternal and infant outcomes： There were 24 cases(34.29%)with complete uterine rupture，4 cases(5.71%)with hysterectomy，30 cases(42.86%)with serious postpartum hemorrhage，21 cases(30%)with massive blood transfusion，and no maternal death. Among the 62 newborns，3 cases (4.84%)died intrauterine，1 case(1.61%)was given up rescue by family members，5 cases(8.06%)had severe neonatal asphyxia，7 cases(11.29%)had mild neonatal asphyxia. ④ Compared with scar uterine rupture，non ⁃ scar uterine rupture was more thanpostpartum rupture(66.67% vs. 16.36%，P ＜ 0.05). Postpartum bleeding rate(80.00% vs. 32.73%)，massive transfusion rate(66.67% vs. 20.00%)and hospitalization cost(26 348.68 yuan vs. 13 859.53 yuan)were significantly increased(P ＜ 0.05)，but there was no significant difference in neonatal outcomes between the two groups(P ＞ 0.05). ⑤ In scarred uterus，the rate of complete uterine rupture (55%)，postpartum hemorrhage(60%)and massive transfusion(45%)in TOLAC group were significantly higher than those in ERCS group(20.00%，17.14%，5.71%，P ＜ 0.05). ⑥ There was no significant difference in maternal and infant outcomes of patients with rupture before and during labor(P ＞ 0.05). However，the rates of complete uterine rupture(26.19% ，11.11%)，massive blood transfusion(16.67%，11.11%)and hospitalization costs(11 576.33 RMB，14 846.30 RMB)were lower than those of postpartum group (63.16%，68.42%，25 310.57 RMB，P ＜ 0.05). Conclusion：In this study，the clinical manifestations of uterine rupture were mainly characterized as abdominal pain，abnormal fetal heart rate or vaginal bleeding，and some patients had no specific clinical manifestations. Non⁃scar uterine rupture，especially if detected late，may result in a more severe adverse outcome.

Abstract:Objective：To compare the advantages and disadvantages of pulmonary ultrasound and fiberoptic bronchoscopy in the localization of endobronchial blocker. Methods：90 patients undergoing elective left thoracotomy for esophageal cancer were randomly divided into two groups：with 45 in each. After anesthesia induction，7.5 cm reinforced tracheal tube was inserted orally，and then the endobronchial blocker was placed blindly to the unilateral bronchus. The position of blocker was located and the effect of pulmonary isolation was judged respectively by pulmonary ultrasonography(group L)and fiberoptic bronchoscopy(group B). Patients were placed in right lateral position and then the blocker position was judged again. The total time needed to judge the pulmonary isolation in group L and group B，the times of intraoperative blocker adjustment，the satisfaction score of intraoperative pulmonary collapse，and the levels of MAP，HR and peak airway pressure(PAW)before and during the two positioning were recorded. Results：There was no significant difference in lung isolation time，lung collapse satisfaction score and intraoperative cuff adjustment times between group L and group B(P ＞ 0.05). Compared with those before positioning，HR，MAP and PAW in group L during the blocker localization had no significant changes(P ＞ 0.05). The levels of HR，MAP and PAW in group B during twice blocker positioning were significantly higher than those before fiberoptic bronchoscope implantation(P ＜ 0.05). Conclusion：The pulmonary isolation effect of endobronchial blocker judged by pulmonary ultrasound is similar to that of fiberoptic bronchoscope，but the effect of pulmonary ultrasound on hemodynamics is less than that of fiberoptic bronchoscopy.

Abstract:Objective：To explore the application value of SonoVue and Sonazoid contrast ⁃enhanced ultrasonography in focal liver lesions. Methods：The clinical and contrast ⁃enhanced ultrasound imaging data of the patients with focal liver lesions in our hospital from January 2020 to December 2021 were analyzed retrospectively. The perfusion pattern，enhancement characteristics of arterial phase，portal phase，delayed phase and late vascular phase(Kupffer phase)of the contrast medium were analysed. Taking the pathological examination results as the“gold standard”，the two examination methods were compared and analyzed. Results：Taking the pathological examination results as the“gold standard”，the accuracy，sensitivity，specificity，positive predictive value and negative predictive value of SonoVue and Sonazoid contrast ⁃enhanced ultrasonography was 88.33%，90.48%，83.33%，92.68%，78.95% and 93.33%，95.24%，88.89%，95.24%，88.89% respectivelv. The kappa value of the consistency test between the two kinds of contrast⁃ enhanced ultrasonography was 0.73. The consistency was at a medium level and the accuracy of SonoVue and Sonazoid contrast ⁃ enhanced ultrasonography in the diagnosis of focal liver lesions was not statistically significant(P=0.373). Conclusion：Sonazoid and SonoVue contrast ⁃ enhanced ultrasonography have no significant difference in the diagnostic accuracy of focal liver lesions，but compared with SonoVue，Sonozoid has a higher quality image，and has the ability to preliminarily judge the differentiation degree of liver tumors，and can also provide a sufficient time for the scanning of other space occupying lesions of the whole liver. Therefore， Sonazoid contrast⁃enhanced ultrasonography has higher clinical application value and broader application prospects.

Abstract:Objective：To establish and validate a prediction model combined machine learning with radiomics features in predicting outcome after mechanical thrombectomy in acute stroke. Methods：Imaging data of acute stroke patients in Nanjing First Hospital were retrospectively collected. These patients were divided into a training set(n=105)and a test set(n=50)according to random number table method. Acute stroke(n=45)in the Second People’s Hospital of Changzhou were collected as the validation set. A.K. software was used to extract radiomics features on diffusion weighted imaging(DWI)and perfusion weighted imaging(PWI). Least absolute shrinkage and selection operator(LASSO)regression model was used to screen the features，and then，the selected features were used to establish the prediction model by support vector machine(SVM)classifier. Receiver operating characteristic(ROC)curve was used to evaluate the predictive efficacy of the model，and the validation set was used to verify the generalization ability of the model. Results： One thousand three hundred and sixteen radiomics features of each patient were extracted from DWI and PWI，and 40 features highly related to outcome after mechanical thrombectomy in acute stroke were screened after dimension reduction. ROC analysis showed that the area under curve(AUC)of DWI+PWI model(training set：0.981；test set：0.891)was higher than those of DWI or PWI model，and the accuracy were 0.943 and 0.900，respectively. The results of validation of the model showed that the prediction model based onDWI + PWI was also better than that of single sequence(DWI or PWI)，the sensitivity and specificity were 0.864 and 0.783 respectively，and the accuracy was 0.822. Conclusion：The prediction model combined machine learning and radiomics can effectively predict outcome after mechanical thrombectomy in acute stroke，and has good generalization ability.

Abstract:Acute intestinal ischemic is a common and critical illness，such as superior mesenteric artery embolism. Ischemia reperfusion injury(IRI)is common after treatment. In the pathological process of intestinal IRI，mitophagy is a“doublE-edged sword”. Its role mainly depends on the degree of IRI. In the pathological changes of mild intestinal IRI，it plays a role in alleviating IRI，while in severe cases，it plays the opposite role. The regulation mechanism of mitophagy in intestinal IRI involves multiple signal pathways， such as PINK1/Parkin signal pathway，BNIP3/NIX signal pathway，FUNDC1 signal pathway and Beclin1 signal pathway. With the deepening of related research，the function and mechanism of mitophagy in intestinal IRI is gradually clear，which provides a reference for accurate intestinal IRI.

Abstract:Traditional oral materials are mostly stable materials which are not affected by oral environment，while intelligent materials can adapt to various changes in the oral cavity and respond favorably，so they have become one of the research hotspots in recent years. As an environmentally sensitive polymer，intelligent hydrogel is an important type of smart materials and has been widely studied in stomatology. Based on the literature at home and abroad，this paper briefly reviews the definition and classification of intelligent hydrogels and the application of intelligent hydrogels in stomatology，and prospect the future development of these materials.