OBJECTIVES: To investigate the antioxidative mechanism of snake venom-derived protein C activator (PCA) in mitigating vascular endothelial cell injury. METHODS: Human umbilical vein endothelial cells (HUVECs) were cultured in DMEM containing 1.0 g/L D-glucose and exposed to hypoxia (1% O₂) for 6 h followed by reoxygenation for 2 h to establish a cell model of oxygen-glucose deprivation/reoxygenation (OGD/R). The cell model was treated with 2 μg/mL PCA alone or in combination with 2-ME2 (a HIF-1α inhibitor) or DMOG (a HIF-1α stabilizer), and intracellular production of reactive oxygen species (ROS) and protein expression levels of HIF-1α, BNIP3, and Beclin-1 were detected using DCFH-DA fluorescence probe, flow cytometry, and Western blotting. The OGD/R cell model was transfected with a BNIP3-specific siRNA or a scrambled control sequence prior to PCA treatment, and the changes in protein expressions of HIF-1α, BNIP3 and Beclin-1 and intracellular ROS production were examined. RESULTS: In the OGD/R cell model, PCA treatment significantly upregulated HIF-1α, BNIP3 and Beclin-1 expressions and reduced ROS production. The effects of PCA were obviously attenuated by co-treatment with 2-ME2 but augmented by treatment with DMOG (a HIF-1α stabilizer). In the cell model with BNIP3 knockdown, PCA treatment increased BNIP3 expression and decreased ROS production without causing significant changes in HIF-1α expression. Compared with HUVECs with PCA treatment only, the cells with BNIP3 knockdown prior to PCA treatment showed significantly lower Beclin-1 expression and higher ROS levels. CONCLUSIONS Snake venom PCA alleviates OGD/R-induced endothelial cell injury by upregulating HIF-1α/BNIP3 signaling to suppress ROS generation, suggesting its potential as a therapeutic agent against oxidative stress in vascular pathologies.
Humans ; Reactive Oxygen Species/metabolism* ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Human Umbilical Vein Endothelial Cells/drug effects* ; Membrane Proteins/metabolism* ; Proto-Oncogene Proteins/metabolism* ; Up-Regulation ; Cell Hypoxia ; Cells, Cultured ; Snake Venoms/chemistry* ; Beclin-1

OBJECTIVE To investigate the effect of bs-5-YHEDA iron chelating peptide combined with semaglutide on the cognitive ability and pathological characteristics of D-Gal-induced Alzheimer's disease(AD) model mice. METHODS Forty mice were randomly divided into 5 groups, namely the healthy control group, PBS group, bs-5-YHEDA iron chelating peptide group, combined treatment group and positive control group, with 8 mice in each group, half of each sex. Except for the healthy control group, D-galactose was injected to induce the AD mice model for 6 weeks. For 3 consecutive weeks starting from the 4th week, the bs-5-YHEDA iron chelating peptide group was injected with bs-5-YHEDA(1 mg·mL–1) once every other day at 200 µL in the tail vein; the bs-5-YHEDA iron chelating peptide(1 mg·mL–1) and semaglutide(25 nmol·kg–1·d–1) were given alternately once a day in the combination treatment group; the positive control group was given memantine(3.3 mg·kg–1·d–1) by gavage every other day. The healthy control group and PBS group were injected with the equal dose of PBS. At the end of treatment, the learning memory ability of mice was detected by the Morris water maze method, whole brain and whole blood were dissected, and pathological changes in hippocampal region were observed by HE staining, and Aβ expression and Tau protein phosphorylation levels were detected by immunohistochemistry, enzyme-linked immunosorbent assay and immunoblotting. RESULTS In the Morris water maze spatial exploration experiment, the differences in the number of times the mice traversed the platform, the ratio of swimming distance to the target quadrant, and the time ratio were statistically significant in each group(P<0.05); compared with the PBS group, the ratio of swimming distance to the target quadrant increased in the combined treatment group, and the differences were statistically significant(P<0.05). The results of HE staining showed that compared with the healthy control mice, the hippocampal area in the PBS group showed reduced levels of pyramidal cells, disorganized arrangement, cell edema, and deep staining of nuclei consolidation. Cellular disorganization, deep staining of nuclei and apoptosis in the hippocampus were significantly improved in each treatment group after drug treatment. Immunohistochemistry and Western blotting results showed that the Aβ expression levels and Tau protein phosphorylation levels were significantly higher in the PBS-administered mice compared with the healthy control mice, and the Aβ expression levels and Tau protein phosphorylation levels were reduced in each group after drug treatment, with statistically significant differences(P<0.01 or P<0.001 ). CONCLUSION The combination of bs-5-YHEDA iron chelating peptide and semaglutide can effectively improve the learning and memory ability and pathological characteristics of AD mice, but from the results of immunohistochemistry and immunoblotting experiments, the improvement of pathological characteristics of AD mice in the combination treatment group is not obvious compared with the single bs-5-YHEDA iron chelating peptide group, suggesting that there may be a threshold effect of our designed dual-target combination treatment on the cognitive improvement of AD mice, and the optimization and validation of the effect of multi-target combination treatment need further study.

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

Objective:To study the correlation between serum N-terminal pro-brain natriuretic peptide (NT-proBNP), homocysteine (Hcy) and SYNTAX score, and further clarify the prognostic significance of NT-proBNP and Hcy in patients with acute myocardial infarction (AMI).Methods:The clinical data of 303 patients with AMI from January 2014 to January 2016 in Lianjiang People′s Hospital of Guangdong Province were retrospectively analyzed. Patients were evaluated using version 2.11 of the SYNTAX score system. The SYNTAX score ≤ 22 scores was divided into the low risk group (103 cases), SYNTAX score 23 to 32 scores was divided into the medium risk group (85 cases), and the SYNTAX score ≥ 33 scores was divided into the high risk group (115 cases). The clinical data and laboratory examination results were collected. Univariate and multivariate Logistic regression analysis were used to analyze the influencing factors of SYNTAX score high risk. The optimal cutoff value of NT-proBNP and Hcy evaluating the SYNTAX score high risk was analyzed by receiver operating characteristic (ROC) curve. Kaplan-Meier survival curve was plotted to compare all-cause mortality of patients with different evaluation indicators. The influencing factors of prognosis in patients with AMI were analyzed by multivariate Cox risk regression analysis.Results:The patients were followed up for 0.1 to 4.5 (4.30 ± 0.76) years, The all-cause death was in 28 cases, and the all-cause mortality rate was 9.2% (28/303). There were statistical difference in age, smoking rate, diabetes mellitus rate, family history rate of coronary artery disease (CAD), left ventricular ejection fraction (LVEF), ST-elevation myocardial infarction (STEMI) rate, white blood cell, fasting blood glucose (FBG), estimation glomerular filtration rate (eGFR), troponin T (TnT), NT-proBNP and Hcy among the 3 groups ( P<0.01 or <0.05). SYNTAX score was positively correlated with NT-proBNP and Hcy ( r = 0.622 and 0.699, P<0.01). Multivariate Logistic regression analysis result showed that diabetes mellitus, LVEF, TnT, NT-proBNP and Hcy were the independent risk factors of SYNTAX score high risk in patients with AMI ( P<0.01 or <0.05). ROC curve analysis result showed that the area under curve (AUC) of NT-proBNP evaluating the SYNTAX score high risk was 0.807 (95% CI 0.757 to 0.850, P<0.05), the optimal cutoff value was 1 816 μg/L, with a sensitivity of 81.03% and a specificity of 82.26%; the AUC of Hcy evaluating the SYNTAX score high risk was 0.743 (95% CI 0.689 to 0.791, P<0.05), the optimal cutoff value was 17.55 μmol/L, with a sensitivity of 66.67% and a specificity of 79.84%. Kaplan-Meier survival curve analysis result showed that the all-cause mortality in high risk group was significantly higher than that in low risk group and medium risk group (17.4% vs. 2.9% and 5.9%), the all-cause mortality in patients with NT-proBNP> 1 816 μg/L was significantly higher than that in patients with NT-proBNP ≤ 1 816 μg/L (16.2% vs. 5.6%), the all-cause mortality in patients with Hcy>17.55 μmol/L was significantly higher than that in patients with Hcy ≤ 17.55 μmol/L (14.2% vs. 5.7%), and there were statistical differences ( P<0.01 or <0.05). Multivariate Cox risk regression analysis result showed that age, diabetes mellitus, SYNTAX score, TnT, NT-proBNP (>1 816 μg/L) and Hcy (>17.55 μmol/L) were the independent influencing factors of all-cause mortality in patients with AMI ( HR = 1.530, 1.368, 2.065, 1.414, 1.821 and 1.510; 95% CI 1.108 to 2.113, 1.012 to 3.485, 1.810 to 2.356, 1.248 to 2.714, 1.606 to 2.064 and 1.278 to 1.783; P<0.05 or <0.01). Conclusions:NT-proBNP>1 816 μg/L and Hcy>17.55 μmol/L can not only reflect the degree, severity and complexity of coronary atherosclerosis, but also predict the prognosis in patients with AMI.

Objective To evaluate the effect of preoperative oral rehydration on postoperative recovery in patients undergoing laparoscopic radical resection of colorectal cancer.Methods Eighty patients of both sexes,aged 18-64 yr,with body mass index of 18-24 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective radical resection of colorectal cancer,were randomly divided into either routine fluid replacement group (group R) or oral rehydration group (group O) with 40 patients in each group.Fasting for solid food was performed at 8 h before surgery and for water at 6 h before surgery,and fluid was replaced according to the volume expansion during induction and 4-2-1 formula in group R.In group O,multivitamin drinks containing sugar and electrolyte was given orally according to the fasting time:12-15 ml/kg at 8 h of fasting,15-20 ml/kg at 8-12 h of fasting,with the total volume not exceeding 1 200 ml,300 ml at 3 h before surgery,and the rest of fluid was given at different times the night before surgery,and fluid replacement 1 200 ml was performed according to the basic requirement of patients and intraoperative fluid loss.Fluid replacement was maintained at a rate of 1.5 ml · kg-1 · h-1 on the day of surgery in two groups.The volume of oral fluid intake,intraoperative net volume of fluid intake and volume of intravenously given fluid on the day of surgery were recorded.The time of surgery,emergence time,time to first flatus,time to first liquid diet,first ambulation time and length of postoperative hospital stay were recorded.The development of postoperative nausea and vomiting and hypotension was recorded.Peripheral venous blood samples were collected on the morning of day 2 after admission to hospital,the day of surgery and day 1 after surgery (T0-2) for determination of the fasting blood glucose and insulin concentrations,and insulin resistance index was calculated.Results Thirty-seven and 35 patients were included in R and O groups,respectively.Compared with group R,the intraoperative net volume of fluid intake and volume of intravenously given fluid on the day of surgery were significantly decreased,fasting blood glucose and insulin concentrations and insulin resistance index were decreased,and the time to first flatus,time to first liquid diet and length of postoperative hospital stay were shortened in group O (P＜0.05).Conclusion Preoperative oral rehydration can promote postoperative recovery,which may be related to mitigating insulin resistance in patients undergoing laparoscopic radical resection of colorectal cancer.

OBJECTIVETo study genetic mutations and clinical features of a pedigree affected with MYH9-related disorders from Guangxi.

METHODSBlood platelets were counted with a hemocytometer. Blood smear was carried out to detect the inclusion body in peripheral blood neutrophils. DNA and mRNA samples were extracted from blood samples from the members of the pedigree. Fragments of the MYH9 gene were amplified with PCR and directly sequenced.

RESULTSThe affected individuals presented with a triad of giant platelets, decreased platelet count and inclusion bodies in the neutrophils with variable expressivity. A heterozygous deletional mutation (c.5803delG) in exon 41 of the MYH9 gene was found in all of the 8 affected individuals, which led to a frame-shift and change of 26 amino acids at the C-end of the tail domain of nonmuscle myosin heavy chain IIA (NMMHC-IIA) (p.Ala1935Profs*12). The same mutation was not found among healthy members of the pedigree.

CONCLUSIONThe c.5803delG mutation probably underlies the MYH9-related disorders in this pedigree. The mutation has altered the C-end of the tail domain of the NMMHC-IIA protein, resulting in mild clinical symptoms in the affected individuals.

Adult ; Base Sequence ; China ; Female ; Humans ; Male ; Molecular Motor Proteins ; genetics ; Molecular Sequence Data ; Myosin Heavy Chains ; genetics ; Pedigree ; Sequence Deletion ; Thrombocytopenia ; diagnosis ; genetics

Objective To study cinical features of patients with idopathic submitral left ventricular aneurysm(ISLVA) and evaluate their global and segmental systolic function as well as diastolic function through echocardiography.Methods Clinical features of eight patients with ISLVA were analysed retrospectively.Standard 2-dimentional and real-time 3-dimentional echocardiography were performed in all the eight cases and other twenty subjects with normal left ventricular(LV) function (defined as control group).Results Two patients were diagnosed as ISLVA due to ventricular arrythmia and the other six case sowing to congestive heart failure.Coronary angiography was normal in all patients.Apical systolic murmur was audible in 6 cases.Four patients received mitral valve surgical repair,one underwent aneurysm resection and radiofrequency ablation,implantable cardioverter defibrillator was implanted in one case,one was only treated by medication.One case died.Patients with ISLVA demonstrated significanly larger left atrium(LA) and LV diameter (both end-diastolic and end systolic),thinner LV posterior wall,and lower LV ejection fraction (LVEF) than controls (P ＜0.05).Indexes of the LV 17 segments time-volume curves including the time to minimal systolic volume(Tmsv) 16-SD,Tmsv 12 SD,Tmsv 6-SD,Tmsv 16-Dif,Tmsv 12-Dif,Tmsv 6-Dif,Tmsv 16-SD％,Tmsv 12-SD％,Tmsv 6-SD％,Tmsv 16-Dif％,Tmsv 12-Dif％,Tmsv 6-Dif％ were significantly higher in patients with ISLVA than those in controls(all P ＜0.05).All patients with ISLVA showed mitral regurgitation and decreased LV diastolic function in varying degrees,five patients accompanied by elevated LA pressure.Conclusions Clinical features of patients with ISLVA are nonspecific.Echocardiography can evaluate systematically their functional and structural abnormalities.

To observe a PPAR-alpha agonist effect of N-oleoylethanolamine (OEA) on CB2 (cannabinoid receptor 2), an anti-inflammatory receptor in vascular endothelial cell, healthy HUVECs and TNF-alpha induced HUVECs were used to establish a human vascular endothelial cell inflammatory model. Different doses of OEA (10, 50 and 100 micromol x L(-1)) had been given to HUVECs, cultured at 37 degrees C for 7 h and then collected the total protein and total mRNA. CB2 protein expression was detected by Western blotting and CB2 mRNA expression was assayed by real-time PCR. As the results shown, OEA (10 and 50 micromol x L(-1)) could induce the CB2 protein and mRNA expression, but not 100 micromol x L(-1). To detect if anti-inflammation effect of OEA is partly through CB2, CB2 inhibitor AM630 was used to inhibit HUVEC CB2 expression, then the VCAM-1 expression induced by TNF-alpha was detected, or THP-1 adhere to TNF-alpha induced HUVECs was examined. OEA (50 micromol x L(-1)) could inhibit TNF-alpha induced VCAM-1 expression and THP-1 adhere to HUVECs, these effects could be partly inhibited by a CB2 inhibitor AM630. The anti-inflammation effect of OEA is induced by PPAR-alpha and CB2, suggesting that CB2 signaling could be a target for anti-atherosclerosis, OEA have wide effect in anti-inflammation, it may have better therapeutic potential in anti-inflammation in HUVECs, thus achieving anti-atherosclerosis effect.

Determining the approach to spread the knowledge and technology for preventing birth defects using the diffusion of innovations theory.Identifying the target group,orientation messages and promotion guidelines based on related literature,laws and clinical epidemiology studies.Pathways integrating both inter-personal communication and IT have pushed the adoption percentage of innovative knowledge for preventing birth defects up from 0％ to 97 ％ in almost no time,helping the region ranking the first in China.The innovation diffusion model has caught attention of both the government and all the community.Prevention measures against birth defects are accepted community wide,while medical workers are regulated by law in their behaviors of mothers and children healthcare services to set a model for prevention of other diseases.

Background and objective Pre-operative assessment of mitral valve(MV)anatomy is essential to surgical design in patientsundergoing MV repair.Although 2-dimensional(2D)echocardiography provides precise information regarding MV anatomy,RT-3DTEE could increase the understanding of MV apparatus and individual scallop identification.We aimed to investigate the value of RT-3DTEE in MV repair. Methods RT-3DTEE was performed in six patients with mitral valve prolapse(MVP) by using Philips IE33with X7-2t probe.Preoperative RT-3DTEE studies were compared with surgical findings in patients undergoing surgical mitral valverepair,and quantitative evaluation was performed by QLab 6.0 software before and after surgicalmitralvalve repair.Results RT-3DTEE could display dynamic morphology of MV,the location of prolapse,and spatial relation to the surrounding tissue.It couldprovide surgical views of the valves and the valvular apparatus.These resuIts were consistent with surgical findings.The quantitativeevahuation before and after surgical MV repair indicated that anterolateral to posteromedial diameter of annalus.anterior to posteriordiameter of annulus,perimeter of annullus.,and area of annalus in projectionplane were significantlv smaller after operation comparedwith those before operation(P＜0.05).The length of posterior leaflet,,the area of anterior and posterior leaflet,the maximal prolapseheight,the volume of leaflet prolapse and the length of coaptation in projection planewere significantly reduced after operation(P＜0.05).Conclusion RT-3DTEE is a unique new medality for rapid and accurate evaluation of mitral valve prolapse and miwal valverepair.