Objective : To investigate the effects of sinapine on lung tissue inflammation and mucus hypersecretion in asthmatic mice. Methods: Eight-week-old female C57BL/6J mice were randomly divided into Control group, ovalbumin(OVA) group, Sinapine group, and Sinapine+OVA group. The asthmatic mice model were established by intraperitoneal injection of OVA combined with aluminum hydroxide [Al(OH)3] suspension and OVA nasal stimulation. One hour before OVA nasal stimulation, the mice in Sinapine+OVA group and Sinapine group were intraperitoneally injected with sinapine solution, and the mice in OVA group and Control group were treated with the same dose of 0.9% sodium chloride solution. 24 hours after the last OVA stimulation, the inflammation of lung tissue of mice were observed by HE staining; the mucus secretion were evaluated by PAS staining; the mRNA expression levels of Interleukin-4(IL-4), Interleukin-5(IL-5), Interleukin-13(IL-13), tumor necrosis factor-alpha(TNF-α), Mucin 5ac(Muc5ac), and the mRNA of the key genes of Notch pathway such as Notch receptor 1(Notch1), Notch receptor 2(Notch2), Notch receptor 3(Notch3), and hes family bHLH transcription factor 1(Hes1) in lung tissues were detected by real-time fluorescent quantitative PCR(RT-qPCR); the expression levels of Notch1, Notch2, Notch3 and Hes1 proteins were determined by Western blot. Results : Compared with Control group, the inflammation score and PAS score of lung tissues of mice in OVA group increased(P<0.001); the mRNA expression levels of IL-4, IL-5, IL-13, TNF-α, and Muc5ac of mice in OVA group were enhanced(P<0.05); the mRNA and protein expression levels of Notch2, Notch3, and Hes1 of mice in OVA group significantly increased(P<0.001), while there was no significant difference in the mRNA and protein expression levels of Notch1. Compared with OVA group, the inflammation score and PAS score of lung tissues of mice in Sinapine+OVA group decreased(P<0.001); the mRNA expression levels of IL-4, IL-5, IL-13, TNF-α, and Muc5ac of mice in Sinapine+OVA group were reduced(P<0.05); the mRNA and protein expression levels of Notch2, Notch3, and Hes1 of mice in Sinapine+OVA group were downregulated(P<0.05), while there was no significant difference in the mRNA and protein expression levels of Notch1. Conclusion Sinapine can alleviate the lung tissue inflammation and mucus hypersecretion in asthmatic mice, and its mechanism may be related to the inhibition of Notch2/Notch3-Hes1 signal pathway.

Objective To investigate the mechanism of autophagy induced by House dust mites(HDM)on airway epithelial tight junction through β-catenin-Snail signaling pathway.Methods Human bronchial epithelial cells(16HBE)were stimulated with HDM at different time points(0,3,6,12,24,48 h)and different concen-trations(0,40,100,200 μg/mL)to screen the appropriate stimulation concentration and stimulation time.16HBE cells were treated with oxidative stress inhibitor N-acetylcysteine(NAC),autophagy inhibitor 3-methylad-enine(3-MA),HDM,and their combinations.Cells were transfected with mCherry-EGFP-LC3B,Beclin-1-siRNA,and ATG14-siRNA lentivirus and then stimulated with NAC and HDM.Immunofluorescence was used to detect the expression levels of autophagy-related protein LC3B,tight junction-related proteins Occludin,and ZO-1 in airway epithelial cells.The level of reactive oxygen species(ROS)was detected by using DCFH-DA in each group.The protein expression levels of Occludin,ZO-1,LC3B,Beclin-1,ATG5,ATG14,P62,Snail,β-catenin and p-β-catenin were detected by Western blot method.Results Immunofluorescence results showed that compared with the control group,200 μg/mL HDM stimulation induced cellular autophagy,increased the expression level of LC3B protein,and promoted the level of ROS,all with statistical significances(all P<0.05).Compared with the HDM group,the HDM+3-MA,HDM+ATG14-si,and HDM+Beclin-1-si groupsall showed significantincreases in the expression levels of tight junction-related proteins Occludin and ZO-1(P<0.05).The HDM+NAC group demonstrated significant decreases both in the level of ROS andin the expression level of LC3B protein.Western blot results revealed that compared with HDM,3-MA and autophagy protein low-expression beads(Beclin-1-si,ATG14-si)attenuated HDM-induced cellular autophagy(P<0.05),inhibited HDM-induced upregulation of Snail and p-β-catenin expression,and improved HDM-induced decreases in Occludin and ZO-1(P<0.05).Moreover,compared with the HDM group,the NAC+HDM group exhibited significant decreases both in the conversion of LC3BⅠ to LC3BⅡ(P<0.001)in the protein levels of Snail,p-β-catenin,Beclin-1 and ATG14(P<0.01),but significant increases in the protein levels of Occludin and ZO-1(P<0.05).Conclusion HDM affects the tight connections between airway epithelial cells by inducing autophagy,which may be attributed to the β-catenin-Snail signaling pathway.

Objective To investigate the mechanism of autophagy induced by House dust mites(HDM)on airway epithelial tight junction through β-catenin-Snail signaling pathway.Methods Human bronchial epithelial cells(16HBE)were stimulated with HDM at different time points(0,3,6,12,24,48 h)and different concen-trations(0,40,100,200 μg/mL)to screen the appropriate stimulation concentration and stimulation time.16HBE cells were treated with oxidative stress inhibitor N-acetylcysteine(NAC),autophagy inhibitor 3-methylad-enine(3-MA),HDM,and their combinations.Cells were transfected with mCherry-EGFP-LC3B,Beclin-1-siRNA,and ATG14-siRNA lentivirus and then stimulated with NAC and HDM.Immunofluorescence was used to detect the expression levels of autophagy-related protein LC3B,tight junction-related proteins Occludin,and ZO-1 in airway epithelial cells.The level of reactive oxygen species(ROS)was detected by using DCFH-DA in each group.The protein expression levels of Occludin,ZO-1,LC3B,Beclin-1,ATG5,ATG14,P62,Snail,β-catenin and p-β-catenin were detected by Western blot method.Results Immunofluorescence results showed that compared with the control group,200 μg/mL HDM stimulation induced cellular autophagy,increased the expression level of LC3B protein,and promoted the level of ROS,all with statistical significances(all P<0.05).Compared with the HDM group,the HDM+3-MA,HDM+ATG14-si,and HDM+Beclin-1-si groupsall showed significantincreases in the expression levels of tight junction-related proteins Occludin and ZO-1(P<0.05).The HDM+NAC group demonstrated significant decreases both in the level of ROS andin the expression level of LC3B protein.Western blot results revealed that compared with HDM,3-MA and autophagy protein low-expression beads(Beclin-1-si,ATG14-si)attenuated HDM-induced cellular autophagy(P<0.05),inhibited HDM-induced upregulation of Snail and p-β-catenin expression,and improved HDM-induced decreases in Occludin and ZO-1(P<0.05).Moreover,compared with the HDM group,the NAC+HDM group exhibited significant decreases both in the conversion of LC3BⅠ to LC3BⅡ(P<0.001)in the protein levels of Snail,p-β-catenin,Beclin-1 and ATG14(P<0.01),but significant increases in the protein levels of Occludin and ZO-1(P<0.05).Conclusion HDM affects the tight connections between airway epithelial cells by inducing autophagy,which may be attributed to the β-catenin-Snail signaling pathway.

Objective:To discuss the effect of Aspergillus fumigatus(Af)on DNA damage and interleukin(IL)-33 expression in the human bronchial epithelial cells,and to clarify its related mechanism.Methods:Different concentrations(1,5,and 10 mg·L-1)of Af were used to stimulate the bronchial epithelial BEAS-2B cells to select the appropriate stimulation concentration.When the BEAS-2B cells were treated with N-acetylcysteine(NAC)and Af,the cells were divided into control group,Af group,NAC group,and Af+NAC group.When the BEAS-2B cells were treated with DNA double-strand break repair inhibitor NU7441 and Af,the cells were divided into control group,Af group,NU7441 group,and Af+NU7441 group.The comet assay was used to detect the percentages of comet tail DNA of cells in various groups;immunofluorescence method was used to detect the expression levels of DNA damage-related protein phosphorylated H2AX(yH2AX)in the cells in various groups;2,7-dichlorofluorescein diacetate(DCFH-DA)fluorescence probe was used to detect the levels of reactive oxygen species(ROS)in the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of interleukih-33(IL-33),thymic stromal lymphopoietin(TSLP),and interleukih-25(IL-25)mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of phosphorylated nuclear factor κB(p-NF-κB),phosphorylated ataxia telangiectasia mutated(p-ATM),and γH2AX proteins in the cells in various groups.Results:Compared with control group,the percentage of comet tail DNA and the expression level of γH2AX in the cells in 1 mg·L-1 Af group showed no significant difference(P＞0.05),while the percentage of comet tail DNA and the expression level of γH2AX in the cells in 5 mg·L-1 Af group were significantly increased(P＜0.01);compared with 5 mg·L-1 Af group,the percentage of comet tail DNA and the expression level of γH2AX in the cells in 10 mg·L-1 Af group were significantly increased(P＜0.01).Compared with control group,the ROS levels in the bronchial epithelial cells in 1 mg·L-1 Af group was significantly increased(P＜0.05);compared with 1 mg·L-1 Af group,the ROS level in the cells in 5 mg·L-1 Af group was significantly increased(P＜0.01);compared with 5 mg·L-1 Af group,the ROS level in the cells in 10 mg·L-1 Af group was significantly increased(P＜0.05).After treatment of NAC,compared with Af group,the percentage of comet tail DNA(P＜0.01),the expression level of γH2AX(P＜0.05),and the ROS level(P＜0.01)in the cells in Af+NAC group were significantly decreased;after treatment of NU7441,compared with Af group,the percentage of comet tail DNA and the expression level of yH2AX in the cells in Af+NU7441 group were significantly increased(P＜0.01).The RT-qPCR results showed that after treatment of NAC,compared with control group,the expression level of IL-33 mRNA in the cells in Af group was significantly increased(P＜0.05);compared with Af group,the expression level of IL-33 mRNA in the cells in Af+NAC group was significantly decreased(P＜0.05);after treatment of NU7441,compared with Af group,the expression level of IL-33 mRNA in the cells in Af+NU7441 group was significantly increased(P＜0.05).The Western blotting results showed that after treatment of NAC,compared with control group,the expression levels of p-NF-κB,p-ATM,and γH2AX proteins in the cells in Af group were significantly increased(P＜0.05);after treatment of NU7441,compared with Af group,the expression levels of p-NF-κB,p-ATM,and γH2AX proteins in the cells in Af+NAC group were significantly decreased(P＜0.05);After treat ment of NU7441,compared with Af group,the expression levels of p-NF-κB,p-ATM,and γH2AX proteins in the cells in Af+NU7441 group were significantly increased(P＜0.05).Conclusion:Af promotes the IL-33 expression in the human bronchial epithelial cells by causing DNA damage,and its mechanism may be related to the activation of ATM/NF-κB signaling pathway.

Objective:To investigate the predictive value of Graeb score for the outcome of high-grade aneurysmal subarachnoid hemorrhage (aSAH) patients with intraventricular hemorrhage (IVH).Methods:Consecutive high-grade aSAH patients with IVH admitted to the No. 1 People's Hospital of Jiujiang from January 2012 to March 2020 were enrolled retrospectively. High-grade aSAH was defined as grade Ⅳ to Ⅴ according to the World Federation of Neurological Surgeons (WFNS) scale. The outcome of patients was evaluated by the modified Rankin Scale (mRS) at 3 months after discharge. A score of ≤2 was defined as a good outcome and a score of >2 were defined as a poor outcome. Multivariate logistic regression model was used to evaluate the correlation between Graeb score and clinical outcome, and the receiver operating characteristic (ROC) curve was used to determine the predictive value of Graeb score for clinical outcome. Results:A total of 86 high-grade aSAH patients with IVH were enrolled. Aneurysm treatment: craniotomy clipping in 42 patients (48.8%), intravascular embolization in 21 (24.4%), and conservative treatment in 23 (26.7%). Twenty-nine patients (33.7%) had a good outcome and 57 (66.3%) had a poor outcome. Multivariate logistic regression analysis showed that the Graeb score >6 (odds ratio [ OR] 26.360, 95% confidence interval [ CI] 4.106-169.235; P<0.001), the modified Fisher grade 3-4 ( OR 11.674, 95% CI 1.540-88.512; P=0.017) and complicated with chronic hydrocephalus ( OR 21.236, 95% CI 2.883-156.431; P=0.003) were the independent risk factors for the poor outcome. ROC curve analysis showed that the area under the curve of the Graeb score predicting for poor outcome was 0.843 (95% CI 0.760-0.926; P<0.001), the best cut-off value was 6.5, and the corresponding sensitivity and specificity were 71.9% and 86.2%, respectively. Conclusion:The Graeb score is an independent influencing factor affecting the clinical outcome of high-grade aSAH patients with IVH. Graeb score >6.5 had higher predictive value for the poor outcome in such patients.

Objective:To investigate the influencing factors for prognoses of patients with high-grade aneurysmal subarachnoid hemorrhage (aSAH).Methods:A retrospective analysis was performed on clinical data of 104 patients with aSAH, admitted to our hospital from January 2012 to March 2010. According to the modified Rankin scale (mRS) scores 3 months after discharge, these patients were divided into good prognosis group (mRS scores≤2) and poor prognosis group (mRS scores >2). Clinical data of patients from the 2 groups were compared. Multivariate Logistic regression analysis was used to analyze the influencing factors for prognoses.Results:Forty patients (38.5%) were with good prognosis and 64 (61.5%) were with poor prognosis. As compared with patients in the poor prognosis group, patients in the good prognosis group had significantly lower proportion of patients>60 years old, and significantly lower Graeb scores, cast fourth ventricle (CFV) incidence, Fisher grading, blood glucose content, diastolic blood pressure, systolic blood pressure and mean arterial pressure, and statistically higher serum potassium content ( P<0.05). There were significant differences in treatment methods and incidence of complications between the two groups ( P<0.05). Multivariate Logistic regression analysis showed that age>60 years ( OR=13.470, 95%CI: 2.177-83.347, P=0.005), Fisher grading ( OR=6.649, 95%CI: 1.141-38.736, P=0.035), conservative treatment ( OR=6.348, 95%CI: 1.048-38.742, P=0.044), mean arterial pressure at admission ( OR=29.721, 95%CI: 3.177-278.012, P=0.003), serum potassium ( OR=5.296, 95%CI: 1.138-24.653, P=0.034), CFV ( OR=9.855, 95%CI: 1.785-51.456, P=0.008), and chronic hydrocephalus ( OR=19.298, 95%CI: 3.294-113.069, P=0.001) were influencing factors for prognoses of high-grade aSAH patients. Conclusion:Severe aSAH patients with advanced age, high Fisher grading, high mean arterial pressure, low serum potassium content, fourth ventricle casting and chronic hydrocephalus under conservative treatment are more likely to have poor prognosis.

Objective:To explore the influence of prognostic nutritional index (PNI) and controlling nutritional status (CONUT) on the prognosis of patients with multiple myeloma.Methods:Data of 157 patients with multiple myeloma (MM) at the affiliated hospital of Xuzhou medical university from January 2014 to December 2018 were retrospectively evaluated. The operating characteristic (ROC) curve analysis was adopted as the optimal cut-off point. PNI and CONUT were grouped based on the cut-off points of 44.45 and 3.5, respectively, and the differences between age, gender, serum calcium, β 2-microglobulin, serum creatinine, lactate dehydrogenase, and hemoglobin were analyzed. The prognostic factors were analyzed via univariate and Cox multivariate regression analyses. Results:The level of PNI and CONUT is the influencing factor of OS time. The univariate analysis revealed that age, LDH, plasma cell ratio, β 2-microglobulin, ISS stage, PNI, and CONUT were the risk factors for the prognosis of patients with MM. The multivariate analysis revealed that age ( HR=1.636, 95% CI 1.014-2.640) , plasma cell ratio ( HR=1.953, 95% CI 1.232-3.096) , and PNI ( HR=0.513, 95% CI 0.287-0.917) were the independent prognostic risk factors of patients with MM. Conclusion:Low PNI in patients with MM indicates a poor prognosis, which is an independent prognosis risk factor.

Objective To investigate the effects of minimally invasive intracranial hematoma clearance on the perihematomal glutamate(Glu) level,permeability of blood-brain barrier(BBB) and brain edema.Methods Thirty rabbits with body weight of 2.80-3.40 kg were used to established the model of spontaneous intracerebral hemorrhage(ICH) and randomly divided into the minimally invasive group(MI) and control group(MC) after the model was prepared successfully.The MI group underwent minimally invasive procedures for removing intracranial hematoma by stereotactic instrument within 6 h after establishing the ICH model.The brain tissue was extracted on postoperative 1,3,7 d,and the perihematomal brain tissues were taken to detect the Glu level,BBB permeability and water content of brain tissue,which were compared with those in the control group.Results The Glu level,BBB permeability and brain water content on 1,3,7 d in the MI group were lower than those in the MC group,and the differences were statistically significant(P＜0.05).Conclusion The minimally invasive surgery for removing intracranial hematoma is helpful to reduce perihematoma Glu level,BBB permeability and brain water content.

BACKGROUND:Crowe type-IV developmental dysplasia of the hip presented complete dislocation of the femoral head, significantly shortening lower limb, serious adverse development of upper segment of acetabulum and femur, and even deformity. At present, there are no reports concerning its therapeutic strategy.
OBJECTIVE:To explore the therapeutic strategy of adult bilateral Crowe type-IV developmental dysplasia of the hip.
METHODA total of 12 cases of adult bilateral Crowe type-IV developmental dysplasia of the hip, who were treated in the Department of Orthopedics, The First People’s Hospital of Changzhou from January 2000 to January 2013, were col ected. There were 5 males and 7 females, at the age of 19-47 years old, averagely 33.5 years old. After two or three weeks of skeletal traction, they received bilateral total hip arthroplasty. Hip rotation center was reconstituted at the level of the“true”acetabulum. The subtrochanteric osteotomy was performed for patients with reduction difficulty. For patients with femoral neck anteversion>40°, subtrochanteric rotary osteotomy was performed.
RESULTS AND CONCLUSION:Disappearance of hip pain, satisfied move function and normal gait were gained after operation. Lower limbs recovered to be isometric. The average leg lengthening was 3.1 cm (range, 2.5 to 4.8 cm). One patient affected sciatic nerve irritation. The average fol owed-up time was 3.5 years. No fracture, dislocation or loosening of the prosthesis appeared. The post-operative Harris Hip Score averaged 86.3. By reconstruction of the hip rotation center in the“true”acetabular level, correction of femoral neck anteversion, reconstruction of function of the abduction muscle and proper selection of prosthesis, total hip arthroplasty for adult bilateral Crowe type-IV developmental dysplasia of the hip could obtain good curative effects.

Objective To establish a multi-drug resistant model of temporal lobe epilepsy and observe the effect of hippocampal stimulation on pharmacoresistant epileptic rats and its possible mechanism with the use of indexes including after discharges (AD) of the amygdalae,the stimulus-induced seizures and the extracellular levels of γ-aminobutyrate (GABA).Methods Totally,120 Wistar rats were used for the amygdaloid kindled model of epilepsy by chronic stimulation of amygaloid basal lateral nucleus.Based on the successful kindled model of epilepsy,we selected the pharmacoresistant and pharmacosensitive epileptic rats according to their response to phenobabital and phenytoin.We then divided the pharmacoresistant epileptic rats into the hippocampal stimulation group and control group,with 8 rats in each group.Low-frequency stimulus was conducted for two weeks in the hippocampal stimulation group.The hippocampus extracellular fluid collected by microdialysis was then used to determine the levels of GABA by a high performance liquid chromatography method after hippocampal stimulation.Results The stimulus-induced seizures were inhibited significantly,and the frequency of the AD was decreased,as well as the amplitude,compared with the control group.The extracellular level of GABA in the 8:00-9:00 am and the 8:00-9:00 pm was (32.69 ± 7.80) and (35.76 ± 6.27) μg/ml,respectively,both significantly increased as compared with the control ((26.58 ± 6.87) μg/ml,t =-21.45,P =0.000 ; (31.50 ± 4.87) μg/ml,t =-15.74,P =0.000).Conclusions The low-frequency hippocampal stimulation inhibits the seizures of the kindled model of epilepsy and decreases the frequency,the amplitude,as well as the duration of the amygdale AD in the pharmacoresistant epileptic rats,which may be contributed to the increased GABA content in extracellular fluid of the brain after stimulation.