Migraine is a common chronic neurological disease，faces still certain limitations in the design of some clinical trials of migraine in China. Based on the principle of “PICO” in clinical research， this article summarizes the core contents of Guidelines for controlled trials of preventive treatment of episodic migraine in adults published by International Headache Society and its Clinical Trials Standing Committee， including subject selection， key points of trial design， and endpoint evaluation indicators. This article also discusses the deficiencies in current clinical trial design， so as to provide a basis for the design of clinical trials for migraine.

Objective: To investigate the effect of phosphatase and tensin homolog(PTEN) overexpression on autophagy in mouse primary cardiac fibroblasts. Methods: Neonatal mice(1-3 days old) were purchased and subjected to cardiac tissue harvesting. Cardiac fibroblasts were isolated through enzymatic digestion and cultured. After cellular adhesion, rapamycin(Rapa)-treated cardiac fibroblasts were used to establish an autophagy model. Following successful model construction, cells were transfected with eitherPTEN-overexpressing plasmid(PTENoverexpression group) or empty vector(control group), followed by 24-48 h incubation. The molecular expressions of PTEN, autophagy-related proteins [microtubule-associated protein light chain 3(LC3Ⅱ/Ⅰ), Beclin1], and fibrotic marker collagen Ⅰ were detected by Western blot and RT-qPCR. Autophagic flux was assessed using mCherry-GFP-LC3 fluorescence staining to evaluate changes in autophagic activity. Transmission electron microscopy(TEM) was employed to observe autophagosome formation in cardiac fibroblasts. Results : In the Rapa-induced cardiac fibroblast autophagy model, compared with the control group, the protein and mRNA levels of PTEN significantly decreased(P<0.05), while the expression of autophagy-related proteins(LC3Ⅱ/Ⅰ, Beclin1) and fibrotic marker Collagen Ⅰ was upregulated at both protein and mRNA levels(P<0.05). Additionally, in PTEN-overexpressing cardiac fibroblasts, the expression levels of LC3Ⅱ/Ⅰ, Beclin1, and Collagen Ⅰ were markedly reduced compared to the empty vector group(P<0.05). mCherry-GFP-LC3 fluorescence staining demonstrated that autophagic activity was significantly attenuated in thePTENoverexpression group versus the empty vector group(P<0.05). TEM further revealed a decreased number of autophagosomes in PTEN-overexpressing cardiac fibroblasts(P<0.05). Conclusion The overexpression ofPTENsignificantly inhibits autophagy in cardiac fibroblasts, suggesting thatPTENmay be a key gene involved in regulating autophagy in cardiac fibroblasts.

AIM:To investigate the preventive and therapeutic effects of sesamin(SES)on fatty liver disease in rats with hypertension combined with dyslipidemia,and to explore the potential mecha-nisms based on mRNA-seq.METHODS:Spontane-ously hypertensive rats(SHRs)were fed a high-fat,high-cholesterol diet to establish a rat model of hy-pertension combined with dyslipidemia,and then treated with SES for 16 weeks continuously.The ex-periment was divided into four groups:WKY,SHR,Model,and Model+SES(160 mg·kg-1·d-1).Blood pressure was measured using the tail-cuff method.Body weight was monitored,and body mass index was calculated.Liver morphology was detected by ultrasound,and liver thickness was measured.Liver wet weight was weighed,and liver index was calcu-lated.Liver volume was detected by the water dis-placement method.Serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT),aspartate amino-transferase(AST),and total bile acids(TBA)were de-tected by ELISA.Liver sequencing analysis was per-formed using mRNA-seq.Liver histomorphological changes were observed by HE staining.The degree of hepatic steatosis was observed by Oil Red O stain-ing,and the degree of hepatic fibrosis was observed by MASSON staining.The mRNA expression of Al-dh1a7,Nnmt,Irs2,Pltp,and Scd was detected by q-PCR.The protein expression of Scd,Nnmt,AMPK,p-AMPK,PPARα,and PPARγ was detected by Western blotting.RESULTS:After 16 weeks of continuous SES administration to rats with hypertension combined with dyslipidemia,blood pressure was significantly reduced(P＜0.01),and body weight was decreased.Serum TG,TC,and LDL-C levels were decreased,while HDL-C levels were increased.Serum ALT and AST levels were decreased.Liver weight,organ in-dex,liver thickness,and liver volume were de-creased.The degree of hepatic steatosis and hepat-ic fibrosis was improved.A total of 545 differentially expressed mRNAs were identified in the livers of rats in each group,of which 278 were upregulated and 267 were downregulated.Among the 27 com-monly differentially expressed mRNAs,five mRNAs related to lipid metabolism were screened,namely Aldh1a7,Nnmt,Irs2,Pltp,and Scd.KEGG enrich-ment analysis showed that the enriched pathways were AMPK and PPAR.Further validation revealed that in the SES-treated group,the mRNA expression of Scd in the liver was decreased,while the mRNA expression of Nnmt was increased.The protein ex-pression of Scd was decreased,while the protein ex-pression of Nnmt,AMPK,p-AMPK,PPARα,and PPARγ was increased.CONCLUSION:SES has preven-tive and therapeutic effects on fatty liver disease in rats with hypertension combined with dyslipidemia,and its mechanism of action may be related to the reduction of Scd expression levels in the liver and the increase in the expression of Nnmt,AMPK,p-AMPK,PPARα,and PPARγ.

Objective To investigate the impact of variations in volute cross-sectional area on the flow characteristics and hemolytic performance of centrifugal blood pumps by designing six volute structures.Methods Computational fluid dynamics and the Lagrangian method were used to analyze flow characteristics and predict hemolysis in blood pumps with different volute designs.Results The annular volute pump showed the poorest hydraulic performance,while the hydraulic performance of the S-shaped volute was the best,which was improved by 35.29％compared to that of the annular volute.Some volutes experienced stagnation zones at the helical inlet(0°-90°)and significant backflow at the outlet(270°-360°).The downward concave-shaped volute had the highest hemolysis index(HI),i.e.,9.59×10-4.Meanwhile,the HI of the annular volute was the lowest,which was 71.85％less than the concave-shaped volute.Conclusions Reducing the gradient of the area variation at the helical inlet and outlet can prevent flow stagnation and backflow.A higher HI arises due to the prolonged exposure of red blood cells to high shear stress.This study provides a theoretical basis for designing and optimizing volute structures of centrifugal blood pumps.

Objective To investigate the impact of variations in volute cross-sectional area on the flow characteristics and hemolytic performance of centrifugal blood pumps by designing six volute structures.Methods Computational fluid dynamics and the Lagrangian method were used to analyze flow characteristics and predict hemolysis in blood pumps with different volute designs.Results The annular volute pump showed the poorest hydraulic performance,while the hydraulic performance of the S-shaped volute was the best,which was improved by 35.29％compared to that of the annular volute.Some volutes experienced stagnation zones at the helical inlet(0°-90°)and significant backflow at the outlet(270°-360°).The downward concave-shaped volute had the highest hemolysis index(HI),i.e.,9.59×10-4.Meanwhile,the HI of the annular volute was the lowest,which was 71.85％less than the concave-shaped volute.Conclusions Reducing the gradient of the area variation at the helical inlet and outlet can prevent flow stagnation and backflow.A higher HI arises due to the prolonged exposure of red blood cells to high shear stress.This study provides a theoretical basis for designing and optimizing volute structures of centrifugal blood pumps.

AIM:To investigate the preventive and therapeutic effects of sesamin(SES)on fatty liver disease in rats with hypertension combined with dyslipidemia,and to explore the potential mecha-nisms based on mRNA-seq.METHODS:Spontane-ously hypertensive rats(SHRs)were fed a high-fat,high-cholesterol diet to establish a rat model of hy-pertension combined with dyslipidemia,and then treated with SES for 16 weeks continuously.The ex-periment was divided into four groups:WKY,SHR,Model,and Model+SES(160 mg·kg-1·d-1).Blood pressure was measured using the tail-cuff method.Body weight was monitored,and body mass index was calculated.Liver morphology was detected by ultrasound,and liver thickness was measured.Liver wet weight was weighed,and liver index was calcu-lated.Liver volume was detected by the water dis-placement method.Serum triglycerides(TG),total cholesterol(TC),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT),aspartate amino-transferase(AST),and total bile acids(TBA)were de-tected by ELISA.Liver sequencing analysis was per-formed using mRNA-seq.Liver histomorphological changes were observed by HE staining.The degree of hepatic steatosis was observed by Oil Red O stain-ing,and the degree of hepatic fibrosis was observed by MASSON staining.The mRNA expression of Al-dh1a7,Nnmt,Irs2,Pltp,and Scd was detected by q-PCR.The protein expression of Scd,Nnmt,AMPK,p-AMPK,PPARα,and PPARγ was detected by Western blotting.RESULTS:After 16 weeks of continuous SES administration to rats with hypertension combined with dyslipidemia,blood pressure was significantly reduced(P＜0.01),and body weight was decreased.Serum TG,TC,and LDL-C levels were decreased,while HDL-C levels were increased.Serum ALT and AST levels were decreased.Liver weight,organ in-dex,liver thickness,and liver volume were de-creased.The degree of hepatic steatosis and hepat-ic fibrosis was improved.A total of 545 differentially expressed mRNAs were identified in the livers of rats in each group,of which 278 were upregulated and 267 were downregulated.Among the 27 com-monly differentially expressed mRNAs,five mRNAs related to lipid metabolism were screened,namely Aldh1a7,Nnmt,Irs2,Pltp,and Scd.KEGG enrich-ment analysis showed that the enriched pathways were AMPK and PPAR.Further validation revealed that in the SES-treated group,the mRNA expression of Scd in the liver was decreased,while the mRNA expression of Nnmt was increased.The protein ex-pression of Scd was decreased,while the protein ex-pression of Nnmt,AMPK,p-AMPK,PPARα,and PPARγ was increased.CONCLUSION:SES has preven-tive and therapeutic effects on fatty liver disease in rats with hypertension combined with dyslipidemia,and its mechanism of action may be related to the reduction of Scd expression levels in the liver and the increase in the expression of Nnmt,AMPK,p-AMPK,PPARα,and PPARγ.

Objective To analyze the clinical characteristics and influencing factors of non-small cell lung cancer (NSCLC) patients with chronic obstructive pulmonary disease (COPD) in Hubei province, and to provide a theoretical basis for the diagnosis and treatment of NSCLC patients with COPD. Methods A total of 246 NSCLC patients admitted to our hospital from 2018 to 2020 were selected and divided into control group (without COPD, n=125) and observation group (with COPD, n=121) according to COPD. The clinical characteristics of chest pain, hemoptysis, emasculation, atelectasis and pleural effusion were compared between the two groups. The values of FEV1/FVC, RV/TLC and DLCO in the two groups were measured by pulmonary function detector. The age, gender, smoking, smoking history, proportion of lung squamous cell carcinoma, TNM stage and other clinical data of all subjects were analyzed by self-made survey scale of our hospital. Univariate analysis and logistic regression were used to analyze the risk factors of COPD in NSCLC patients. Results Among 246 NSCLC patients, 121 patients (49.19%) were complicated with COPD, including 76 males and 45 females, and there was a statistical difference between the two groups (χ²=4.891, P>0.05). The average age of the observation group (61.02±4.82) was significantly higher than that of the control group (59.76±4.73) (t=2.069, P<0.05). The proportion of chest pain, hemoptysis, emaciation, atelectasis, pleural effusion and fatigue in the observation group were significantly higher than those in the control group (P<0.05). The values of FEV1/FVC, RV/TLC and DLCO in the observation group were significantly lower than those in the control group (P<0.05). There were significant differences in smoking history, proportion of lung squamous cell carcinoma and TNM score between the two groups (P>0.05). Male (OR=2.982), smoking history (OR=2.623) and lung squamous cell carcinoma (OR=3.147) were risk factors for COPD in NSCLC patients (P<0.05). Conclusions NSCLC patients with COPD are more common in male smokers in Hubei Province, often accompanied by pleural effusion , severe hemoptysis and other symptoms , and their lung function is decreased. Early detection and standardized treatment of COPD in the treatment of NSCLC can improve the prognosis of patients.

Objective To explore the role of secreted frizzled-related protein 3 (sFRP3), a regulator of the Wnt signaling pathway, in the activation and proliferation of murine cardiac fibroblasts (CFs).Methods Neonatal mice aged 1-3 days were obtained for surgical procedures to collect heart tissues.After digestion, CFs were isola-ted and cultured.Transforming growth factor-beta 1 (TGF-β1) stimulation was used to induce activation and prolif-eration in CFs after they adhered to the culture dish.Once the model was confirmed, experimental and control groups were transfected with sFRP3 overexpression plasmids and empty plasmids for 24-48 hours.Expression lev-els of sFRP3, Periostin (POSTN), Type Ⅰ collagen (Collagen Ⅰ), and proliferating cell nuclear antigen (PC-NA) were assessed at the molecular level using Western blot and qRT-PCR.Changes in cell proliferation capacity were examined using MTT, CCK-8, and EdU staining methods.Results In the TGF-β1-induced activation and proliferation model of CFs, compared to the control group, the model group exhibited decreased expression of sFRP3 protein and mRNA, while the expression of activation and proliferation-related proteins PCNA, POSTN, and Collagen Ⅰ was upregulated.Furthermore, in CFs overexpressing sFRP3 through plasmid transfection, the protein and mRNA expression of PCNA, POSTN, and Collagen Ⅰ decreased compared to the empty vector group.MTT, CCK-8 , and EdU experiments indicated a significant decrease in the proliferative activity of CFs in the sFRP3 over-expression group compared to the empty vector group.Conclusion Overexpression of sFRP3 markedly inhibits the activation and proliferation of CFs, suggesting that sFRP3 may be a key gene involved in the regulation of CF acti-vation and proliferation.

Objective:To investigate the effects and significance of acute and chronic trauma on brain degree centrality (DC) in patients with post-traumatic stress disorder (PTSD) who lost their only child at resting state.Methods:Retrospectively, the study enrolled a total of 51 parents with PTSD, including 35 PTSD parents whose children was lost in emergencies (acute bereaved PTSD group) and 16 PTSD parents whose children was lost of chronic causes such as diseases (chronic bereaved PTSD group). Fifty local adults were also included as healthy controls (HC group). The clinical administered PTSD scale(CAPS) was used to evaluate the severity of the subjects' clinical symptoms.Resting-state functional magnetic resonance imaging(fMRI) data of all subjects were collected and DC values were calculated.SPSS 22.0 software was used for statistical analysis.Covariance analysis was performed among three groups, while post hoc was performed between any two groups.What's more, correlation analyses were utilized between abnormal brain regions and the scores of CAPS.Results:Significant group effects were found in multiple regions, including the right inferior temporal gyrus (MNI: x, y, z=66, -27, -21), right temporal pole (MNI: x, y, z=54, 15, -9), right orbital inferior frontal gyrus (MNI: x, y, z=42, 21, -15), bilateral medial superior frontal gyri (MNI: right x, y, z=6, 63, 12; left x, y, z=-3, 60, 18), left inferior parietal angular gyrus (MNI: x, y, z=-45, -36, 51) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51). Compared with HC group, the DC of two patient groups increased in the right inferior temporal gyrus (MNI: acute x, y, z=63, -27, -21; chronic x, y, z=63, -21, -27); the DC of acute bereaved PTSD group decreased in the right temporal pole (MNI: x, y, z=45, 21, -15) and the right orbital inferior frontal gyrus (MNI: x, y, z=48, 24, -12), while the DC of chronic bereaved PTSD group decreased in the left inferior parietal angular gyrus (MNI: x, y, z=-45, -36, 51) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51). Compared with chronic bereaved PTSD group, the DC of acute bereaved PTSD group increased in the left inferior parietal angular gyrus (MNI: x, y, z=-33, -39, 42) and left postcentral gyrus (MNI: x, y, z=-45, -33, 51), while decreased in the right temporal pole (MNI: x, y, z=51, 12, -9), right orbital inferior frontal gyrus (MNI: x, y, z=42, 21, -15) and bilateral medial superior frontal gyri (MNI: left x, y, z=0, 57, 15; right x, y, z=3, 57, 15). In chronic bereaved PTSD group, the DC of the left postcentral gyrus was negatively correlated with C1 (avoid trauma-related thoughts, feelings) score in CAPS ( r=-0.606, P=0.028). In acute bereaved PTSD group, the DC of the left medial superior frontal gyrus was negatively correlated with D4 (high vigilance) score ( r=-0.416, P=0.020). Conclusion:There exist functional abnormalities of multiple brain regions in acute and chronic bereaved parents with PTSD.The high arousal symptoms of the former may be related with the abnormalities of prefrontal-amygdala neural circuit, while the latter show higher avoidance which may be associated with the dysfunction of somatosensory brain regions such as postcentral gyrus.

Objective:To study the clinical features and molecular diagnosis of neonates with Beckwith-Wiedemann syndrome (BWS).Methods:From January 2016 to December 2020, infants admitted to the Department of Neonatology of Children's Hospital of Fudan University with BWS diagnosed during the neonatal period were reviewed. Their clinical data including clinical features, molecular testing results and comorbidities of other congenital diseases were analyzed.Results:A total of 16 BWS neonates (9 males and 7 females) were included, with average gestational age of 34~39 weeks and average birth weight of 2 412~4 275 g. Macroglossia (9/16) and omphalocele (8/16) were the main clinical features. Birth weight >2 SD of the mean (12/16) was the common secondary feature. According to the European BWS diagnostic scoring criteria, 10 infants with a ≥4 score were clinically diagnosed with BWS. 5 infants with a 2~4 score and 1 infant with a 1 score (whose twin brother had a 4 score) were clinically suspected BWS.13 infants received molecular testing. 6 infants with clinical diagnosis and 5 infants with suspected diagnosis were confirmed to have reduced methylation signal in imprint control region 2 and 2 of the clinically diagnosed infants had increased methylation signal in imprint control region 1. 5 infants had atrial septal defect and brain injury were also seen in 5 infants. Abnormal thyroid function, abnormal umbilical cord morphology and abnormal umbilical blood vessels were seen in 4 infants, respectively.Conclusions:The combination of the clinical features and molecular testing can establish the diagnosis of BWS as early as during the neonatal period. Due to the high risk of comorbidities, abdominal ultrasonography, cardiac ultrasonography, cranial MRI and thyroid function tests are necessary after the diagnosis of BWS.