ABSTRACT:Objective To explore role of U0126,the specific inhibitor of ERK signaling pathway,in early brain injury (EBI)and the autophagy of nerve cells in hippocampus area in subarachnoid hemorrhage (SAH). Methods A total of 48 male adult SD rats were randomly divided into control group,SAH group,DMSO+SAH group,and U0126+SAH group,with 12 in each.We established SAH rat model by the puncture of internal carotid artery.The same amount of saline water,DMSO and U0126 solution of 0.5 mL per rat was injected respectively into the rats of different groups 30 min before modeling.The rats were killed at 24 h.To measure brain water content by Wet and dry method after 24 h,the morphological changes of hippocampus CA1 neural cells were observed by microscopy;the expression levels of ERK,Beclin-1 and LC3 were detected by using immunohistochemical method. Results Compared with that in sham group,brain water content increased obviously in SAH model group.The density of surviving neurons in SAH group was significantly lower than that in control group (P<0 .0 5 ).ERK signaling pathway was activated obviously,the expressions of Beclin 1 and LC3-Ⅱ were significantly higher than those in control group (P<0.05).Compared with SAH model group,in U0126 group brain water content increased obviously.Compared with those in SAH group,the density of surviving neurons was significantly lower (P<0.05), ERK signaling pathway was suppressed,the expressions of Beclin-1 and LC3-Ⅱ were significantly lower (P<0.05). Conclusion The U0126,the ERK signaling pathway inhibitor,can inhibit neuron autophagy and increase EBR of SAH.

Objective To investigate the incidence and related risk factors of delirium in elderly patients with hip frac-ture. Methods From October 2014 to February 2017, 306 patients with femoral neck fracture or intertrochanteric fracture more than 65 years old underwent elective hip arthroplasty (hip arthroplasty, proximal femoral nail fixation) were retrospective-ly analyzed. Patients' general information was recorded: age, sex, body mass index (BMI), history of diabetes mellitus, history of coronary heart disease, cerebral infarction, preoperative biochemical tests, left ventricular ejection fraction, thrombosis of both lower extremities, intraoperative status (ASA score, NYHA heart function score, anesthesia, anesthesia time, etc.), periop-erative blood transfusion, biochemical examination on the first day after surgery, perioperative albumin consumption, etc.; ac-cording to whether delirium happened after surgery, those patients were divided into delirium group and non-delirium Group and analyzed the incidence of postoperative delirium. Using univariate logistic regression analysis screened factors associated with postoperative delirium. The results were entered into the multivariate logistic regression analysis, screening the high risk factors for postoperative delirium.Results The incidence of delirium was 59(19.29%) after perioperative hip fracture, includ-ing 23 males and 36 females, ranging in age from 70 to 93 years with an average of (81.9±5.4) years. 247 cases had no deliri-um,including 81 males and 166 females,aged 65-96 years,with an average(76.4±8.1)years.The factors of age,length of hos-pital stay, days before surgery, preoperative hematocrit, perioperative albumin consumption, perioperative blood transfusion, preoperative leukocyte levels, preoperative albumin levels, preoperative creatinine, urea level, Mallampati classification (Mar-kov grade), cardiac function score (NYHA score), ASA score, intraoperative blood loss, postoperative leukocyte level, left ven-tricular ejection fraction, previous cerebral infarction, history of coronary heart disease, and diabetes were statistically signifi-cant different; univariate logistic regression analysis showed that the factors of age, ASA score, history of diabetes mellitus, length of hospitalization, and perioperative albumin consumption were demonstrated to have relationship with the occurrence of postoperative delirium. Multivariate regression analysis showed that age, history of diabetes and ASA score are the high risk factors of postoperative delirium.Conclusion Age,history of diabetes mellitus and ASA score are risk factors of delirium in el-derly patients with hip fracture.Those patients older than 75 years,with the history of diabetes mellitus or ASA score> 2 points were at higher risk of postoperative delirium,which prolongs hospitalization and increases protein consumption.

Objective:To explore the mechanism of extracorporeal shock wave combined with platelet rich plasma in the treatment of articular cartilage injury in knee osteoarthritis (KOA) rats based on the silent information regulator 1 (Sirt1)/forkhead transcription factor O1 (FoxO1) pathway.Methods:15 SD rats were used for platelet rich plasma extraction and 35 SD rats were randomly divided into blank control group, model group, extracorporeal shock wave group, platelet rich plasma group, and extracorporeal shock wave + platelet rich plasma group. Each group had 7 cases. After the intervention, HE staining of articular cartilage tissue was used to observe changes in articular cartilage morphology, Mankin score was used for pathological evaluation, CCK-8 method was used to detect chondrocyte vitality and proliferation, ELISA method was used to detect inflammatory factor levels in joint fluid, and Western Blot method was used to detect the expression levels of Sirt1 and acely-FoxO1/FoxO1 in five groups of articular cartilage tissue.Results:The HE staining of articular cartilage tissue showed that model group, extracorporeal shock wave group, platelet rich plasma group, and extracorporeal shock wave + platelet rich plasma group had varying degrees of pathological damage, with model group having the most severe pathological damage, while the other three experimental groups had no significant differences. The Mankin score and the level of acely-FoxO1/FoxO1 in articular cartilage tissue showed that blank control group < extracorporeal shock wave + platelet rich plasma group < platelet rich plasma group < extracorporeal shock wave group < model group (all P < 0.05). The results of Sirt1 level in articular cartilage tissue, activity, and proliferation ability of articular chondrocytes showed that model group < extracorporeal shock wave group < platelet rich plasma group < extracorporeal shock wave + platelet rich plasma group < blank control group (all P < 0.05). Comparison of inflammatory factor levels in joint fluid, blank control group < extracorporeal shock wave + platelet rich plasma group < extracorporeal shock wave group < platelet rich plasma group < model group (all P < 0.05). Conclusions:The combination of extracorporeal shock wave and platelet rich plasma can promote the proliferation of osteoarthritis chondrocytes and alleviate joint inflammation and cartilage damage in KOA rats by upregulating Sirt1 expression and downregulating FoxO1 acetylation levels.

Objective:To explore the predictive value of platelet-albumin-bilirubin(PALBI)score for tumor recurrence after liver transplantation(LT)in patients with hepatocellular carcinoma(HCC).Methods:Clinical data were retrospectively reviewed for 102 HCC patients undergoing LT from January 2010 to December 2020.The predictive value of PALBI score for tumor recurrence after LT and the risk factors for tumor recurrence after LT were examined by receiver operating characteristic(ROC)curve, Kaplan-Meier method and univariate/multivariate Cox regression.Results:The optimal cutoff value of preoperative PALBI score for predicting recurrence was -3.82 with ROC curve, Youden's index 0.317 and area under the ROC curve 0.679.Survival analysis was performed using a PALBI cutoff value of -3.82 as boundary group.The results showed that significant differences existed in 1/3/5-year tumor recurrence rates(17.9% vs.50.0%, 26.9% vs.62.5%, 29.5% vs.62.5%)after low PALBI and high PALBI( P<0.05 for all). Univariate analysis indicated that preoperative tumor maximal diameter, tumor number, Milan criteria, alpha fetoprotein(AFP)level, microvascular invasion, portal venous tumor thrombus, and PALBI score were significantly associated with postoperative tumor recurrence( P<0.05 for all). And multivariate analysis revealed that Milan criteria, AFP level and PALBI score were independent risk factors for postoperative tumor recurrence( P<0.05). Conclusions:Preoperative PALBI score offers some predictive value for postoperative tumor recurrence in HCC patients post-LT.When preoperative PALBI score ≥-3.82 in HCC patients, postoperative tumor recurrence rate is relatively high.

Objective: To study the relationship between motor coordination, social communication impairment, and restricted/repetitive behaviors (RRBs) in children with autism spectrum disorder (ASD). Methods: A total of 123 ASD children aged 2-12 were recruited from rehabilitation service institutions for autistic children in Guangzhou, including 105 boys and 18 girls. The Developmental Coordination Disorder Questionnaire, Social Communication Questionnaire, and Restrictive Behavior Scale Revised were collected to measure motor coordination, social communication skills and RRBs, respectively. Multiple linear regression was used to explore the association between motor coordination and core symptoms. Results: In children aged 2-5 years old, those without motor coordination deficits scored lower in social interaction (7.73±4.00) than others (9.76±2.90)( Z=-2.29, P <0.05). In children aged 6-12 years old, those without motor coordination deficits scored lower in restricted, repetitive, and stereotyped patterns of behavior (4.22±1.93), repetitive sensory-motor behaviors (3.91±2.63), and insistence on sameness (13.52±10.59) than those with motor coordination deficits (6.11±1.54, 14.00±8.49, 31.33±15.05)( Z=-2.48, -3.83, -3.28, P <0.01)。Multiple linear regression analysis showed that fine motor/handwriting and control during movement were negatively correlated with the severity of social communication impairment and RRBs respectively among children aged 2-5 and 6-12( β=-0.85--0.03, -2.98--0.17, P <0.05). Conclusion Among autistic children, those with motor deficits suffered from more severe core symptoms. Motor assessment should be included in the ASD screening, diagnosis and management. Motor intervention should also be promoted in the rehabilitation in order to improve the prognosis and life quality.

Objective To investigate the role and mechanism of nico-tinamide adenine dinucleotide phosphate oxidase 4 (NAPHD oxidase 4,Nox4)-mediated reactive oxygen species (ROS) generation on high-dose dexamethasone (DEX) induced apoptosis in osteoblasts.Methods According to culture conditions,3rd passage of murine osteoblastic MC3T3-E 1 cells were divided into control group,Dexamethasone group,Dexamethasone+NAC (N-acetyl-L-cysteine) group,NAC group,Dexamethasone+DPI (Diphenyleneiodonium) group and DPI group.24 hours after culture,the morphology of osteoblasts was observed by inverted phase contrast microscope.Cell viability was determined by MTT assay.The generation of ROS in osteoblasts was measured using a fluorescent probe DCFH-DA.The apoptosis of each group was observed through Hoechst staining.The mRNA level and protein expression of Nox4 were detected by real-time quantitative PCR and Western Blot.In addition,after silence of Nox4 with small interfering RNA (siRNA),the ROS generation was further detected by a fluorescent probe DCFH-DA.Results After treatment with 1000 nmol DEX for 24 hours,compared to control group,the results of inverted phase contrast microscope and MTT showed that osteoblasts in DEX group exhibited more obvious signs of shrinkage and deformation with decreased cell viability.After intervene with NAC and DPI,morphology of osteoblasts was good with increased viability of osteoblasts.Compared to control group (5.86％± 0.28％),the production of ROS in DEX group (45.14％±1.49％) was significantly increased (P=0.000).The apoptotic rates in DEX group (29.60％± 1.52％) was significantly increased compared with control group (4.12％±0.67％) (P=0.000).Compared to DEX group,the production of ROS generation in DEX+NAC group (28.06％±1.61％) and DEX+DPI group (23.70％±1.28％) was significantly decreased (P=0.000).It presented that NAC or DPI significantly decreased the formation of ROS.Compared to DEX group,the apoptotic rate in DEX+NAC group (8.94％± 1.47％) and DEX+DPI group (12.96％±2.03％) was significantly decreased (P=0.000).It presented that NAC or DPI significantly decreased osteoblast apoptosis.In addition,the Nox4 mRNA level in DEX group was 2.67-fold compared with control group (t=-10.301,P=0.009).The difference had statistically significance.The protein expression of Nox4 in DEX group was 2.37-fold compared with control group (t=-15.542,P=0.004).The difference has statistically significance.After silence of Nox4 by siRNA,the generation of ROS in DEX+Nox4 siRNA group (14.53％± 1.00％) was decreased by 16.92％ compared with DEX group 31.45％±0.72％ (P=0.000).The difference had statistically significance.Conclusion Nox4-mediated ROS generation plays an important role in osteoblasts apoptosis induced by high-dose dexamethasone.It provided us the new target in the management of Nox4 to provide possible therapy for steroid-induced avascular necrosis of the femoral head (SANFH).

Objective:To evaluate the outcomes of aortic root repair in acute type A aortic dissection (ATAAD) with aortic sinus involvement.Methods:The clinical data of patients with ATAAD involving the aortic sinus and an aortic root diameter of ≤45 mm who were treated from January 1, 2015, to December 31, 2016, were collected. Patients were divided into group A (involvement of one aortic sinus or part of one aortic sinus) and group B (involvement of more than one aortic sinus). The effectiveness of aortic root repair in ATAAD with sinus involvement was analyzed by comparing the preoperative imaging characteristics and postoperative results between the two groups.Results:The study cohort comprised 155 patients, including 100 patients in group A and 55 in group B. There were no differences between the two groups in baseline characteristics, aortic root diameter, 30-day mortality, and complication rates. During an average follow-up of(49.1±14.6) months, there was no difference between the two groups in the survival rate (92.0% vs. 89.5%, P=0.61). The aortic root diameter at follow-up showed no progressive expansion compared with preoperatively in either of the two groups[(38.1±3.6) mm vs. (37.9±3.5)mm, P=0.92, A; (38.4±4.1) mm vs. (38.3±3.6) mm, P=0.74, B]; furthermore, there was no difference between the two groups in aggravation of aortic regurgitation. Conclusion:Aortic root repair achieves satisfactory medium-term outcomes in ATAAD with more than one aortic sinus involved, but the long-term outcomes need to be evaluated.

Objective: To investigate the association between parenting style and sleep problems among school aged children with autism spectrum disorder (ASD). Methods: A total of 98 children with ASD aged 6-10 years old and 98 age and gender matched typically developing (TD) children from mainstream schools were recruited. Parenting style and sleep problems were measured via Parent Behavior Inventory (PBI) and Children s Sleep Habits Questionnaire(CSHQ), respectively. The symptom severity and intelligence level were also evaluated. Generalized linear model was used to analyze the relationship between parenting style and sleep problems. Results: There was no statistically significant difference in the parenting style of the two groups of children( P > 0.05 ); weekend sleep time of children with ASD was significantly shorter than that of the TD group [(9.1±0.7)(9.5±0.8)h, P < 0.01 ], and the score of sleep onset delay was significantly higher than that of the TD group[(1.8±0.7)(1.5±0.7), P <0.01]. However, there was no statistically significant difference in the incidence of total sleep problems and various problems between the two groups of children( P >0.05). The parental support/engagement of children with ASD was negatively associated with the total score of sleep problems( β=-2.68, 95%CI =-4.88--0.47), bedtime resistance ( β=-1.65, 95%CI =-2.54--0.77) and sleep anxiety( β=-1.01, 95%CI =-1.70--0.32). The parental hostility/coercion was positively correlated with score of daytime sleepiness( β=1.41, 95%CI =0.53-2.29)( P <0.05). Conclusion Parenting style of support/engagement is associated with lower sleep problems in children with ASD, while hostile/coercion is associated with higher sleep problems. It should be improve parental style to reduce the sleep problems in children with ASD.

Objective: To discuss the relationship between lingual tonsil hypertrophy and laryngopharyngeal reflux. Methods: Ninety-two patients who received throat surgery in Nanfang Hospital between October 2015 and October 2016 were enrolled. Twenty-six healthy volunteers were recruited as normal controls. All participants were assessed with the reflux finding score(RFS) and the size of lingual tonsils were evaluated using a clinical grading system proposed by Friedman under electronic laryngoscope. The score of reflux symptom index(RSI), personal history and medical history were gathered. Biopsy specimens of lingual tonsils were taken from all participants for the immunohistochemical stain of pepsin.SPSS 19.0 software was used for statistical analysis. Results: There were 46.2% (12/26) pepsin-positive and 53.8% (14/26) pepsin-negative volunteers in normal controls. There were 87.0% (80/92) pepsin-positive and 13.0% (12/92) pepsin-negative patients in study group. The severity of lingual tonsil hypertrophy and expression intensity of pepsin in patients were significantly higher in volunteers (Z=-3.636, Z=-5.273, P<0.01). The severity of lingual tonsil hypertrophy was positively associated with the pepsin level in patients (r=0.556, P<0.01). The patients with pepsin-positive expression showed significant correlation between lingual tonsil hypertrophy and the positive rate of RSI and RFS (r=0.258, r=0.225, P<0.05). Analysis of correlated factors indicated that lingual tonsil hypertrophy was associated with smoking (χ²=8.502, P<0.05). Conclusions The expression of pepsin can be detected in lingual tonsil tissues. The lingual tonsil hypertrophy is closely related to laryngopharyngeal reflux.

BACKGROUND: Gallbladder cancer (GBC) is the most common malignant tumor of biliary tract. Isoliquiritigenin (ISL) is a natural compound with chalcone structure extracted from the roots of licorice and other plants. Relevant studies have shown that ISL has a strong anti-tumor ability in various types of tumors. However, the research of ISL against GBC has not been reported, which needs to be further investigated. METHODS: The effects of ISL against GBC cells in vitro and in vivo were characterized by cytotoxicity test, RNA-sequencing, quantitative real-time polymerase chain reaction, reactive oxygen species (ROS) detection, lipid peroxidation detection, ferrous ion detection, glutathione disulphide/glutathione (GSSG/GSH) detection, lentivirus transfection, nude mice tumorigenesis experiment and immunohistochemistry. RESULTS: ISL significantly inhibited the proliferation of GBC cells in vitro . The results of transcriptome sequencing and bioinformatics analysis showed that ferroptosis was the main pathway of ISL inhibiting the proliferation of GBC, and HMOX1 and GPX4 were the key molecules of ISL-induced ferroptosis. Knockdown of HMOX1 or overexpression of GPX4 can reduce the sensitivity of GBC cells to ISL-induced ferroptosis and significantly restore the viability of GBC cells. Moreover, ISL significantly reversed the iron content, ROS level, lipid peroxidation level and GSSG/GSH ratio of GBC cells. Finally, ISL significantly inhibited the growth of GBC in vivo and regulated the ferroptosis of GBC by mediating HMOX1 and GPX4 . CONCLUSION ISL induced ferroptosis in GBC mainly by activating p62-Keap1-Nrf2-HMOX1 signaling pathway and down-regulating GPX4 in vitro and in vivo . This evidence may provide a new direction for the treatment of GBC.
Animals ; Mice ; Carcinoma in Situ ; Chalcones/pharmacology* ; Ferroptosis ; Gallbladder Neoplasms/genetics* ; Glutathione Disulfide ; Kelch-Like ECH-Associated Protein 1 ; Mice, Nude ; NF-E2-Related Factor 2/genetics* ; Reactive Oxygen Species ; Humans