Objective To screen differentially expressed miRNAs(DEMs)by comparing the expression of miRNAs in serum exosomes between Alzheimer's disease(AD)patients and healthy controls.Methods A total of 71 AD patients admitted to Department of Geriatric Neurology of Xiangya Hospital from March 2017 to August 2018 and another 71 healthy individuals who taking physical examination in the hospital during same period were recruited and assigned into AD and HC groups,respectively.Four AD patients and four healthy subjects were selected for high-throughput second-generation sequencing of exosome miRNAs.The results were analyzed to obtain the DEMs between them,and the top 4 DEMs were finally selected.Then real-time quantitative real-time PCR was applied for all the subjects to detect the expression of the 4 DEMs.Results High-throughput second-generation sequencing detected 775 miRNAs,and 44 DEMs were found with statistical difference between the 2 groups(P＜0.05).Compared with the HC group,34 miRNAs were up-regulated and 10 were down-regulated in the AD group.The top 4 DEMs were miRNA-148a-3p,miRNA-16-5p,miRNA-19b-3p and miRNA-483-5p.MiRNA-148a-3p was significantly up-regulated in the AD group than the HC group(P＜0.01),but there were no significant differences in the expression level in the other 3 DEMs between the 2 groups.ROC curve analysis showed that the area under the curve of miRNA-148a-3p was 0.7113(95％CI:0.622～0.801),with a sensitivity of 71.6％and a specificity of 69.7％.Conclusion Serum exosome miRNA-148a-3p can be used as a biomarker for the diagnosis of AD.

Objective:To detect the expression of microRNA (miR)-211-5p, erythropoietin hepatocyte kinase receptor B2 (EphB2) and erythropoietin hepatocyte kinase ligand B2 (ephrin B2) in spinal cord tissues as well as nerve cells after spinal cord injury (SCI), and to explore their mechanisms and effects on neurological recovery in SCI rats.Methods:The study was conducted from May 2020 to June 2021 using Sprague Dawley (SD) rats and PC12 cells. SD rats were divided into sham-operated group and SCI group of 30 rats each, and Basso-Beattie-Bresnahan (BBB) score were performed at different postoperative time points (1, 3, 7, 14, 21 and 28 d), and the relative expression of miR-211-5p and Eph/ephrin B2 mRNA was measured by quantitative real-time polymerase chain reaction (qPCR); the SCI rats were divided into recombinant lentiviral vector LV-miR-211-5p group (group A), empty lentiviral vector LV-eGFP (group B) and saline group (group C), with 15 rats in each group, respectively. The recombinant lentiviral vector, empty lentiviral vector and saline were injected on the cephalic and caudal sides of the spinal cord injury, and the relative expression of miR-211-5p and Eph/ephrin B2 mRNA in the spinal cord tissue was measured at 1, 7 and 14 d after surgery. In addition, a PC12 injury cell line model was established with 150 μmol/L hydrogen peroxide (H 2O 2), and the apoptosis rate and apoptosis-related proteins and contents of different cell lines were detected by flow cytometry and Western blot, respectively. MiR-211-5p was verified to target EphB2 by dual luciferase reporter gene. Results:The results of the animal experiments showed that at different postoperative time points, the miR-211-5p levels in the SCI group were lower than those in the SHAM group: 0.70 ± 0.03 vs. 1.00 ± 0.10, 0.60 ± 0.04 vs. 1.00 ± 0.05, 0.45 ± 0.10 vs. 1.00 ± 0.12, 0.30 ± 0.06 vs. 1.00 ± 0.15, 0.20 ± 0.05 vs. 1.00 ± 0.13, 0.10 ± 0.02 vs. 1.00 ± 0.07. In contrast, levels of Eph/ephrin B2 were higher in the SCI group compared to the SHAM group: 1.10 ± 0.05 vs. 1.00 ± 0.01, 1.80 ± 0.01 vs. 1.00 ± 0.08, 2.30 ± 0.01 vs. 1.00 ± 0.10, 2.60 ± 0.01 vs. 1.00 ± 0.05, 2.80 ± 0.01 vs. 1.00 ± 0.06, 3.00 ± 0.01 vs. 1.00 ± 0.07 and 1.20 ± 0.05 vs. 1.00 ± 0.02, 1.60 ± 0.01 vs. 1.00 ± 0.03, 2.10 ± 0.10 vs. 1.00 ± 0.01, 2.40 ± 0.11 vs. 1.00 ± 0.09, 2.70 ± 0.13 vs. 1.00 ± 0.05, 2.90 ± 0.12 vs. 1.00 ± 0.03 ( P<0.05). At 14 d after surgery, Group A exhibited higher BBB scores than Groups B and C: (14.0 ± 1.1) points vs. (8.0 ± 1.1) and (8.2 ± 1.2) points, while miR-211-5p levels were higher than those in Groups B and C: 1.90 ± 0.10 vs. 0.40 ± 0.01 and 0.50 ± 0.02, and Eph/ephrin B2 levels were lower than those in Groups B and C: 0.70 ± 0.10 vs. 1.80 ± 0.04 and 1.90 ± 0.06, 0.60 ± 0.03 vs. 2.00 ± 0.04 and 2.10 ± 0.05 ( P<0.05). Immunofluorescence staining showed that the levels of GAP-43 and synaptophysin in group A were higher than those in groups B and C at 14 d after surgery ( P<0.05). Cellular assays showed that overexpression of miR-211-5p inhibited the apoptosis rate of H 2O 2-induced PC12 cells and the expression of the apoptosis-related gene Cleaved-caspase3 ( P<0.05). Knockdown of miR-211-5p increased the apoptosis rate of H 2O 2-induced PC12 cells and the expression of the apoptosis-related gene Cleaved-caspase3 ( P<0.05). Dual luciferase reporter gene assay confirmed that EphB2 was a target gene of miR-211-5p and overexpression of EphB2 antagonized the inhibitory apoptosis effect of miR-211-5p on H 2O 2-induced PC12 cells. Conclusions:This study showed that miR-211-5p could promote neurological repair in SCI by inhibiting the expression of Eph/ephrin B2 signaling pathway, suggesting that using miR-211-5p as a target to inhibit Eph/ephrin B2 signaling pathway may have a protective effect on SCI.

Methods: (i) Animal experiments. This study conducted experiments using specific pathogen-free (SPF) grade male C57BL/6J wild-type (WT) mice and APP/PS1 double transgenic mice. The animals were divided into three groups: WT group (WT mice, n = 5, receiving distilled water daily), APP/PS1 group (APP/PS1 double transgenic mice, n = 5, receiving distilled water daily), and BSTSD group [APP/PS1 double transgenic mice, n = 5, treated with BSTSD suspension at a dosage of 27 g/(kg·d) for 90 d]. Cognitive function was assessed using the Morris water maze (MWM). Post-experiment, hippocampal tissues were collected for analysis of pyramidal cell and synaptic morphology through hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM). (ii) Cell experiments. The HT-22 cells were divided into control group (untreated), Aβ25-35 group (treated with 20 μmol/L Aβ25-35 for 24 h), icariin group (pre-treated with 20 μmol/L icariin for 60 min, followed by 20 μmol/L Aβ25-35 for an additional 24 h), and icariin + LY294002 group [treated with 20 μmol/L icariin and 20 μmol/L LY294002 (an inhibitor of the phosphoinostitide 3-kinases (PI3K) signaling pathway) for 60 min, then exposed to 20 μmol/L Aβ25-35 for 24 h], and cell viability was measured. Western blot was used to detect the expression levels of synapse-associated proteins [synaptophysin (SYP) and postsynaptic density-95 (PSD-95)] and PI3K signaling pathway associated proteins [phosphorylated (p)-PI3K/PI3K, p-protein kinase B (Akt)/Akt, and p-mechanistic target of rapamycin (mTOR)/mTOR]. Results: (i) Animal experiments. Compared with APP/PS1 group, BSTSD group showed that escape latency was significantly shortened (P < 0.01) and the frequency of crossing the original platform was significantly increased (P < 0.01). Morphological observation showed that pyramidal cells in the hippocampal CA1 region were arranged more regularly, nuclear staining was uniform, and vacuole-like changes were reduced after BSTSD treatment. TEM showed that the length of synaptic active zone in BSTSD treatment group was increased compared with APP/PS1 group (P < 0.01), and the width of synaptic gap was decreased (P < 0.01). (ii) Cell experiments. Icariin had no obvious toxicity to HT-22 cells when the concentration was not more than 20 μmol/L (P > 0.05), and alleviated the cell viability decline induced by Aβ25-35 (P < 0.01). Western blot results showed that compared with Aβ25-35 group, the ratios of p-PI3K/PI3K, p-Akt/Akt and p-mTOR/mTOR in icariin group were significantly increased (P < 0.01), while the protein expression levels of SYP and PSD-95 were increased (P < 0.01). These effects were blocked by LY294002 (P < 0.01). Conclusion BSTSD and icariin enhance cognitive function and synaptic integrity in AD models and provide potential therapeutic strategies through activation of the PI3K/Akt/mTOR pathway.

Objective:To investigate the correlation between thyroid hormone resistance and hyperuricemia in euthyroid population.Methods:It was a cross-sectional study. A total of 548 euthyroid subjects who were hospitalized or underwent physical examination in the Heart Center and Health Management Center of the First Affiliated Hospital of Tsinghua University from January 2021 to December 2022 were selected. Thyroid function, uric acid, blood lipid and other indicators were collected in the subjects, and thyroid function parameters were calculated: thyroid-stimulating hormone index (TSHI), thyrotropin T4 resistance index (TT4RI), parametric thyroid feedback quantile-based index (PTFQI), free triiodothyronine/free thyroxine (FT3/FT4), the sum activity of peripheral deiodinases (SPINA-GD) and the secretory capacity of the thyroid gland (SPINA-GT). According to uric acid level, the subjects were divided into high uric acid group and normal group, the clinical characteristics and thyroid function parameters of the two groups were compared, the correlation between thyroid hormone resistance and hyperuricemia was further explored.Results:Compared with the normal group, male proportion (94.4% and 52.5%), smoking rate (5.2% and 21.3%), body mass index [(27.10±3.96) and (24.26±3.42) kg/m 2], waist-to-hip ratio [0.90(0.86, 0.94) and 0.86(0.80, 0.91)], serum creatinine [(85.50±12.27) and (73.77±28.79) μmol/L], total cholesterol [(5.08±0.99) and (4.72±0.86) mmol/L], triglyceride [2.10(4.40, 5.59) and 1.14(0.79, 1.67)mmol/L], low-density lipoprotein [(3.19±0.97) and (2.85±0.84) mmol/L] and homocystine [(15.07±9.13) and (12.50±10.85) μmol/L] were all higher in the high uric acid group, and the level of high-density lipoprotein [1.15(1.43, 2.88) and 1.39(1.16, 1.64) mmol/L] was lower (all P<0.05). In the aspect of thyroid hormone and thyroid function parameters, the FT4[16.90(5.40,17.95) and 16.00(14.30,17.80) pmol/L], FT3[5.56(5.25, 5.94) and 5.22(4.81, 5.63) pmol/L], FT3/FT4 [0.34(0.31, 0.37) and 0.32(0.29, 0.36)], TSHI (2.70±0.50 and 2.58±0.60), PTFQI (0.406±0.332 and 0.335±0.353) and SPINA-GD [3.72(3.41, 4.05) and 3.52(3.18, 4.00) ]were all higher in high uric acid group than those in normal group(all P<0.05). Spearman correlation analysis showed that blood uric acid level was positively correlated with FT4 ( r=0.185), FT3 ( r=0.422), FT3/FT4 ( r=0.16), TSHI ( r=0.134), TT4RI ( r=0.09), PTFQI ( r=0.121) and SPINA-GD ( r=0.157) (all P<0.05). Conclusion:In people with euthyroid function, central resistance to thyroid hormone is correlated with hyperuricemia.

Objective To compare the early effect of arthroscopic modified Mason-Allen(mMA)and suture-bridge technique for small and medium-size rotator cuff tear.Methods 60 patients with small and medium-size rotator cuff tear were divided into mMA group and suture bridge group,30 cases each.The mMA group was treated with mMA technique,and the suture bridge group was treated with suture-bridge technique.The operative time,the number of anchors used,blood loss,shoulder mobility,pain visual analogue scale(VAS),American Shoulder and Elbow Surgeons(ASES)scale,the Constant-Murley score and the rate of re-tear were compared between the two groups.Results The operative time and the number of anchors used in mMA group were less than those in suture bridge group(P<0.05),and there was no difference of blood loss between the two groups(P>0.05).After operation,the shoulder joint motion of two groups was increased compared with before operation(P<0.05),and there was no statistical difference between the two groups(P>0.05).After surgery,VAS of the two groups was lower than that before surgery,ASES scale and Constant-Murley scores were higher than those before surgery(P<0.05),there was no difference in VAS,ASES and Constant-Murley scores between the two groups(P>0.05).There was no difference in re-tear rate between the two groups(P>0.05).Conclusion Arthroscopic mMA is similar to suture-bridge technique in the treatment of small and medium-size rotator cuff tear,but operative time of mMA is shorter and it requires less anchors.

Objective To investigate the clinical efficacy of endoscopic pterygoid nerve combined with anterior ethmoidal nerve transection in treatment of allergic rhinitis with nasal polyps.Method A clinical practice study was conducted on 108 patients with allergic rhinitis complicated with nasal polyps.After enrollment,they were randomly divided into two groups,54 cases were included in the control group and treated with conventional endoscopic sinus surgery,while 54 cases were included in the observation group and treated with conventional endoscopic sinus surgery combined with pterygoid nerve and ethmoidal nerve transection.The nasal pain visual analogue scale(VAS),rhinoconjunctivitis quality of life questionnaire(RQLQ),asthma quality of life questionnaire(AQLQ),total effective rate,and complication rate of the two groups were compared before treatment and 1 week,2 weeks,3 weeks,3 months,6 months,and 1 year after treatment.Result Compared with the control group,the total effective rate of treatment was 98.15%in the observation group,which was higher than 85.19%in the control group,the difference was statistically significant(P<0.05).Before treatment,there were no significant differences in nasal resistance(NR),mucociliary transport rate(MTR),mucociliary transport time(MTT),and Lund-Kennedy scores between the two groups,the differences were no statistically significant(P>0.05).After treatment,MTR in two groups increased compared with before treatment,and the observation group was higher than the control group,the difference was statistically significant(P<0.05);NR,MTT,and Lund-Kennedy scores in two groups decreased,and the observation group was lower than the control group,the differences were statistically significant(P<0.05).Before treatment,there were no significant differences in nasal pain VAS,RQLQ,and AQLQ between the two groups,the differences were no statistically significant(P>0.05).After treatment,at 1 week,2 weeks,3 weeks,and 3 months,6 months,and 1 year,the observation group had lower nasal pain VAS compared to the control group,while the observation group had lower RQLQ and higher AQLQ compared to the control group,the differences were statistically significant(P<0.05).The incidence of complications in the observation group was lower than the control group,the difference was statistically significant(P<0.05).Conclusion The clinical efficacy of endoscopic pterygoid nerve combined with ethmoidal nerve transection for the treatment of allergic rhinitis complicated with nasal polyps is significant.It can not only effectively alleviate nasal pain in patients,but also improve their quality of life and reduce postoperative complications,which has clinical application value.

Objectives:The present study evaluated the impact of ablation pain during pulmonary vein isolation(PVI)on catheter-tissue contact at different regions. Methods:Forty consecutive patients with atrial fibrillation(AF)referred to Central China Fuwai Hospital for catheter radiofrequency ablation from February to May 2023 were enrolled.The pulmonary veins on each side were divided into 8 regions.The catheter-tissue contact force(CF)and the number of ablation contact stability(>50%catheter attach time CF≥10 g)of each ablation lesion were analyzed.Pain scores during the ablation were assessed using the Faces Pain Scale-Revised and the maximum score was taken for each ablation region.Based on the pain scores,in each region,20 cases with higher pain scores were categorized into the pain group and 20 cases with lower pain scores were categorized into the normal group.The CF characteristics of each region and the relationship with ablation induced pain were analyzed. Results:A total of 3 832 lesions were recorded in 40 patients with AF,with a mean CF of(12.2±7.8)g.Among them,the CF in the pain group was significantly lower than that in the normal group([11.1±5.1]g vs.[13.4±4.8]g,P<0.05).The top region of the right pulmonary vein was the region with the largest CF(16.5±5.8)g,and the upper part of the left anterior wall(at the ridge between the left atrial appendage)was the region with the smallest CF(7.5±3.7)g.At the bottom of right pulmonary vein,right lower posterior wall,left pulmonary vein,and left posterior wall,as well as the middle region of left posterior wall,and upper region of left posterior wall,the CF was significantly smaller in the pain group than that in the normal group(all P<0.05).Of the 3 832 lesions,2 193(57.2%)were stable lesions,and the proportion of stable lesions in the pain group was significantly lower than that in the normal group(55.2%vs.59.5%,P<0.05).In the right pulmonary vein bottom,right lower posterior wall,left lower anterior wall,left pulmonary vein bottom,and left lower posterior wall,the proportion of stable lesions was significantly lower in the pain group than in the normal group(all P<0.05).In addition,the ratio of stable lesions in left pulmonary vein regions was lower than in the right(54.2%vs.60.5%,P<0.05).In the upper part of the left anterior wall(at the ridge between the left atrial appendage),only 88(39.3%)of the 224 lesions in 40 patients were stable lesions. Conclusions:Pain during ablation significantly affects the stability of the catheter to tissue.Monitoring real-time CF during PVI may have important implications for improving ablation efficacy,especially in regions with more intense pain.

DNMT3A encodes a DNA methyltransferase involved in development, cell differentiation, and gene transcription, which is mutated and aberrant-expressed in cancers. Here, we revealed that loss of DNMT3A promotes malignant phenotypes in lung cancer. Based on the epigenetic inhibitor library synthetic lethal screening, we found that small-molecule HDAC6 inhibitors selectively killed DNMT3A-defective NSCLC cells. Knockdown of HDAC6 by siRNAs reduced cell growth and induced apoptosis in DNMT3A-defective NSCLC cells. However, sensitive cells became resistant when DNMT3A was rescued. Furthermore, the selectivity to HDAC6 inhibition was recapitulated in mice, where an HDAC6 inhibitor retarded tumor growth established from DNMT3A-defective but not DNMT3A parental NSCLC cells. Mechanistically, DNMT3A loss resulted in the upregulation of HDAC6 through decreasing its promoter CpG methylation and enhancing transcription factor RUNX1 binding. Notably, our results indicated that HIF-1 pathway was activated in DNMT3A-defective cells whereas inactivated by HDAC6 inhibition. Knockout of HIF-1 contributed to the elimination of synthetic lethality between DNMT3A and HDAC6. Interestingly, HIF-1 pathway inhibitors could mimic the selective efficacy of HDAC6 inhibition in DNMT3A-defective cells. These results demonstrated HDAC6 as a HIF-1-dependent vulnerability of DNMT3A-defective cancers. Together, our findings identify HDAC6 as a potential HIF-1-dependent therapeutic target for the treatment of DNMT3A-defective cancers like NSCLC.

ObjectiveTo validate the alleviating effect of Gegen Qinliantang （GGQLT） on insulin resistance in db/db diabetic mice by regulating the silent information regulator 1 （SIRT1）/forkhead transcription factor O1 （FoxO1） autophagy pathway. MethodSeventy-five SPF-grade spontaneous type 2 diabetic db/db mice and 15 control db/m mice were selected and maintained on regular feed for one week before measuring blood glucose. They were randomly divided into six groups， with 15 mice in each group. The groups included a normal group （physiological saline， 0.2 g·kg^-1）， a metformin group （0.2 g·kg^-1）， high-， medium-， and low-dose GGQLT groups （31.9， 19.1， 6.9 g·kg^-1）， and a model group （physiological saline， 0.2 g·kg^-1）. They were orally treated with corresponding drugs for eight weeks， once daily. Fasting blood glucose （FBG） was measured using a Roche glucometer. Serum levels of high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， triglyceride （TG）， and total cholesterol （TC） were measured using an automated biochemical analyzer. Fasting serum insulin （INS） levels were determined using enzyme-linked immunosorbent assay （ELISA）， and the homeostasis model assessment of insulin resistance （HOMA-IR） was calculated. Western blot was used to detect the expression of Beclin-1， microtubule-associated protein 1 light chain 3 （LC3）， and SIRT1/FoxO1 autophagy pathway-related proteins in liver tissues. Immunohistochemistry was performed to assess the expression of SIRT1， FoxO1， Beclin-1， and LC3B proteins in liver tissues. Transmission electron microscopy was used to observe the formation of autophagosomes in the liver. ResultCompared with the normal group， the model group showed significant increases in FBG， FINS， HOMA-IR， TC， TG， LDL-C， and HDL-C levels （P<0.01）， and significant increases in the expression of SIRT1， Beclin-1， LC3， and FoxO1 proteins in liver tissues （P<0.01）. Transmission electron microscopy revealed the highest number of autophagosomes in the model group. Compared with the model group， the metformin group and the low-， medium-， and high-dose GGQLT groups showed significant decreases in serum FBG， FINS， HOMA-IR， TC， TG， LDL-C， and HDL-C levels （P<0.05， P<0.01）， significant decreases in the expression of SIRT1， Beclin-1， LC3 （P<0.05， P<0.01）， and up-regulated FoxO1 protein （P<0.01）. Transmission electron microscopy showed a reduction in the degree of autophagy in the treatment groups. Compared with the metformin group， the medium- and high-dose GGQLT groups showed significant decreases in FBG， FINS， and TG levels （P<0.01）， significant decreases in the expression of SIRT1， Beclin-1， and LC3 in liver tissues （P<0.05， P<0.01）， and reduced FoxO1 protein （P<0.01）. The high-dose GGQLT group showed reduced HOMA-IR， TC， LDL-C， and HDL-C levels （P<0.05， P<0.01）. Transmission electron microscopy revealed a significant reduction in autophagosomes in the medium- and high-dose GGQLT groups. ConclusionGGQLT can significantly improve glucose and lipid metabolism disorders， alleviate insulin resistance in db/db mice， and prevent and treat type 2 diabetes by activating the SIRT1/FoxO1 autophagy pathway.

Objective:To analyze the significance and factors influencing of CT scan under the modified Valsalva maneuver. Methods:Clinical data of 52 patients with hypopharyngeal carcinoma diagnosed from August 2021 to December 2022 were collected, all patients had calm breathing CT scan and modified Valsalva maneuver CT scan. Compare the exposure effect of the aryepiglottic fold, interarytenoid fold, postcricoid area, piriform fossa apex, posterior hypopharyngeal wall, and glottis with each CT scanning method. The effects of age, neck circumference, neck length, BMI, tumor site, and T stage on the exposure effect were analyzed. Results:In 52 patients, 50 patients（96.15%） completed CT scan at once time. The exposure effect of the CT scan under modified Valsalva maneuver in the aryepiglottic fold, interarytenoid fold, postcricoid area, piriform fossa apex, posterior hypopharyngeal wall was significantly better than CT scan under calm breathing（Z=-4.002, -8.026, -8.349, -7.781, -8.608, all P<0.01）, while CT scan under modified Valsalva maneuver was significantly worse in glottis than CT scan under calm breathing（Z=-3.625, P<0.01）. In the modified Valsalva CT scan, age had no obvious effect on the exposure effect. The exposure effect was better with long neck length, smaller neck circumference, smaller BMI and smaller T stage. The exposure of postcricoid carcinoma was better than pyriform sinus carcinoma and posterior hypopharyngeal wall carcinoma. But differences were not all statistically significant. Conclusion:The anatomical structure of the hypopharynx was clearly under CT scan with modified Valsalva maneuver, which clinical application is simple, but the effect of glottis was worse. The influence of age, neck circumference, neck length, BMI, and tumor T stage on the exposure effect still needs further investigation.
Humans ; Hypopharynx/diagnostic imaging* ; Valsalva Maneuver ; Hypopharyngeal Neoplasms/surgery* ; Tomography, X-Ray Computed ; Carcinoma