Objective To explore the effect of the knockdown of transmembrane 9 superfamily protein member 2 (TM9SF2) on the replication of vesicular stomatitis virus (VSV), and investigate its role in the mechanism of antiviral innate immunity. Methods Small interfering RNA (siRNA) was used to knock down the TM9SF2 gene in human non-small cell lung cancer A549 cells. The CCK-8 method was used to assess cell proliferation. A VSV-green fluorescent protein (VSV-GFP) infected cell model was established. The plaque assay was used to measure the viral titer in the supernatant. RT-qPCR and Western blotting were employed to quantify the mRNA and protein levels of VSV genome replication in A549 cells following VSV infection, as well as the expression of interferon β (IFN-β) mRNA and interferon regulatory factor 3 (IRF3) protein phosphorylation following polyinosinic-polycytidylic acid (poly(I:C)) stimulation. Results Compared to the negative control, the knockdown of TM9SF2 exhibited a significant effect, with no observed impact on A549 cell proliferation. The VSV-GFP infected A549 cell model was successfully established. After viral stimulation, fluorescence intensity was reduced following TM9SF2 knockdown, and the mRNA and protein levels of VSV were significantly downregulated. The viral titer of VSV was decreased. After poly(I:C) stimulation, TM9SF2 knockdown significantly upregulated the mRNA level of IFN-β and the phosphorylation level of IRF3 protein. Conclusion The knockdown of TM9SF2 inhibits the replication of vesicular stomatitis virus, and positively regulates the type I interferon signaling pathway, thus enhancing the host's antiviral innate immune response.
Humans ; Virus Replication/genetics* ; Signal Transduction ; Membrane Proteins/metabolism* ; A549 Cells ; Vesiculovirus/physiology* ; Interferon-beta/metabolism* ; Interferon Regulatory Factor-3/genetics* ; Interferon Type I/metabolism* ; Vesicular Stomatitis/immunology* ; Gene Knockdown Techniques ; Vesicular stomatitis Indiana virus/physiology* ; RNA, Small Interfering/genetics*

Objective To investigate the relationship between cognitive impairment and sleep parameters in acute ischemic stroke patients with obstructive sleep apnea(OSA).Methods A total of 343 patients with acute ischemic stroke and OSA were selected.The cognitive function was assessed using the simple mental state examination scale(MMSE).Patients were divided into the stroke with OSA and cognitive impairment group(MMSE＜27 points,n=119)and the stroke with OSA without cognitive impairment group(MMSE≥27 points,n=224).General data,TOAST etiological classification and distribution of cerebral infarction lesions were collected.The intelligent sleep monitoring system was used to calculate apnea hypopnea index(AHI)and evaluate OSA.Objective sleep monitoring parameters were collected at night.Sleep monitoring was conducted within 24 h of admission and continuous monitoring for≥3 nights.Continuous monitoring duration≥7 h every night to obtain night sleep structure parameters.Multifactor Logistics regression was used to analyze the relationship between cognitive impairment and sleep parameters in stroke patients with OSA.Results Compared with the stroke with OSA without cognitive impairment group,the proportion of age,diabetes history and HHcy history,the proportion of patients with infarct lesions located in frontal,temporal,parietal,occipital,thalamus,basal ganglia,brainstem and hemioval center increased in the stroke with OSA and cognitive impairment group,and the number of years of education decreased,the number of waking times,the proportion of light sleep and AHI increased,the nighttime sleep efficiency and deep sleep period decreased(P＜0.05).Logistics regression analysis showed that after controlling for years of education,age and other interference factors,nighttime sleep efficiency and AHI were strongly associated with cognitive impairment in acute ischemic stroke patients with OSA(P＜0.05).The increased nighttime sleep efficiency was protective factor for cognitive impairment,and increased AHI was risk factor for cognitive impairment.Conclusion Cognitive impairment in acute ischemic stroke patients with OSA is closely related to sleep parameters,in which the increased sleep efficiency at night is a protective factor for cognitive impairment,and the increased AHI is a risk factor.

Objective:To explore the application value of four-dimensional automatic left ventricular quantitation(4D Auto LVQ) technology, in evaluating the myocardial mechanics in patients with different risk stratifications of hypertrophic cardiomyopathy(HCM).Methods:A total of 88 HCM patients and 20 healthy volunteers were selected from February 2020 to February 2022 in the First Affiliated Hospital of Zhengzhou University. According to the HCM Risk-SCD score, HCM patients were divided into 3 groups: low-risk group( n=49), intermediate-risk group( n=21), and high-risk group( n=18). Conventional ultrasound parameters were collected, and 4D Auto LVQ technology was used to obtain the mechanical parameters of left ventricular myocardium, including global longitudinal strain(GLS) , global circumferential strain(GCS), global area strain(GAS), global radial strain(GRS), twist and torsion. The differences in these parameters among the four groups were compared. The predictive values of conventional ultrasound parameters and myocardial mechanical parameters in patients with intermediate- and high-risk HCM patients were analyzed by ROC curve. Results:①Left ventricular end-diastolic diameter, left ventricular end-diastolic volume, left ventricular end-systolic volume, and peak systolic velocity of mitral annulus in the low-, intermediate-, and high-risk groups were lower than those in the control group while left ventricular maximal wall thickness(LVMWT) and early diastolic peak velocity of mitral value orifice/early diastolic peak velocity of mitral annulus(E/e′) were higher, left atrial diameter(LAD) and left ventricular outflow tract gradients(LVOTG) in the intermediate- and high-risk groups were higher than the low-risk group(all P<0.05). ②Compared with the control group, the GLS of HCM patients was lower, and the GLS of the intermediate- and high-risk groups was lower than the low-risk group. GCS and GRS in the intermediate- and high-risk groups were lower than those in the low-risk group. GAS in the high-risk group was lower than the low-risk and the control group, but higher than the intermediate-risk group(all P<0.05). Compared with the control group, the twist and torsion in the intermediate- and high-risk groups were higher, but lower than the low-risk group, and the differences were statistically significant(all P<0.05). ③The ROC results showed that the area under the curve(AUC) of the model containing conventional ultrasound parameters(LVWMT, LAD, and LVOTG) for predicting intermediate- and high-risk HCM patients was 0.811, with a sensitivity of 0.769 and a specificity of 0.755. The AUC of the conventional ultrasound parameters combined with myocardial mechanical parameters was 0.904, as the sensitivity was 0.667 and the specificity was 0.980. Conclusions:4D Auto LVQ can evaluate the mechanical characteristics of LV myocardium in HCM patients with different risk stratifications. Myocardial mechanical parameters combined with conventional ultrasound parameters can improve the diagnostic performance of patients with intermediate- and high-risk HCM.

Objective To evaluate the role of cholinergic anti-inflammatory pathway in dexmedeto-midine pretreatment-induced reduction of acute lung injury in a rat model of intestinal ischemia-reperfusion ( I∕R) . Methods Twenty-four healthy male Sprague-Dawley rats, aged 2-3 months, weighing 200-250 g, were divided into 4 groups ( n=6) using a random number table method: sham operation group ( S group) , intestinal I∕R group ( II∕R group ) , dexmetomidine group ( DEX group) and α7 nicotinic acetyl-choline receptor antagonistα-bungarotoxin (α-BGT) group (α-BGT group) . Intestinal I∕R was produced by occlusion of the superior mesenteric artery ( SMA) for 60 min followed by 120 min of reperfusion in anesthe-tized rats. Dexmetomidine 5 μg·kg-1 ·h-1 was injected via the tail vein at 1 h before operation in DEX group andα-BGT group. α-BGT 1μg∕kg was intraperitoneally injected at 15 min before dexmetomidine in-jection in α-BGT group. Rats were sacrificed at 120 min of reperfusion, and lung tissues were obtained for microscopic examination of pathological changes ( with a light microscope) and for determination of wet∕dry weight ratio ( W∕D ratio) , tumor necrosis factor-alpha ( TNF-α) and interleukin-6 ( IL-6) contents ( using enzyme-linked immunosorbent assay) , malondialdehyde ( MDA) content ( using thiobarbital acid method) and superoxide dismutase ( SOD) activity ( by xanthine oxidase method) . Results Compared with group S, the W∕D ratio and contents of MDA, TNF-αand IL-6 were significantly increased, and the SOD activi-ty was decreased in II∕R and α-BGT groups, and TNF-α and IL-6 contents were significantly increased in group DEX ( P<0. 05) . Compared with group II∕R, the W∕D ratio and contents of MDA, TNF-αand IL-6 were significantly decreased, SOD activity was increased (P<0. 05), and the pathological changes were significantly attenuated in group DEX. Compared with group DEX, the W∕D ratio and contents of MDA, TNF-α and IL-6 were significantly increased, SOD activity was decreased ( P<0. 05) , and the pathological changes were accentuated in group α-BGT. Conclusion Activation of cholinergic anti-inflammatory path-way is involved in the mechanism by which dexmedetomidine pretreatment reduces acute lung injury in a rat model of intestinal I∕R.

Objective: To investigate hepatitis C virus(HCV)genotyping and the serum HCV-RNA concentration in patients infected with different HCV genotypes and to provide information for evaluation of disease condition and anti-viral treatment efficacy. Methods: A total of 60 anti-HCV positive serum samples were collected before antiviral treatment. RT-PCR was performed for the 5′ non-cording region and was followed by nucleotide sequencing for HCV genotyping. Meanwhile, serum HCV-RNA concentration was detected by quantitative PCR. SPSS21.0 and Graphpad Prism 5.0 software were used for data analysis. Analysis of variance (ANOVA) was used for comparison among multi-groups and the t-test was used for comparison between two groups. Results: The frequencies of HCV genotypes 1b, 3a, 1a and 2a were 48.3% (29/60), 23.3% (14/60), 16.7% (10/60) and 10% (6/60), respectively. And, there is one subtype 2c was detected in this study. The mean serum viral concentration with standard deviation of HCV in genotype 1a, 1b, 2a, and 3 a were 5.46±1.19, 6.22±0.78, 5.47±0.65, and 5.38±0.98 log₁₀ (IU/ml) respectively. Conclusions The infection rate of HCV genotype 1 was significantly higher than that of genotype 2 and 3 (P<0.01). The statistical analysis showed the serum HCV-RNA concentration in patients with subtype 1b was significantly higher than that of the subtype group 1 a, 2 a, and 3 a (P<0.05). The study of the relationship between HCV genotypes and the serum HCV-RNA concentration may contribute to anti-viral treatment prescription for hepatitis C patients.

Objective To evaluate the correlation between the types of vertebral artery occlusion and their compensatory hemodynamic changes and posterior circulation ischemia using color Doppler flow imaging combined with transcranial color-coded sonography.Methods From June 2015 to June 2016,A total of 108 patients with vertebral artery occlusion confirmed by vascular sonography,digital subtraction angiography (DSA) or CT angiography (CTA) were enrolled retrospectively.According to the magnetic resonance imaging (MRI)-diffusion weighted imaging (DWI) findings,they were divided into posterior circulation infarction (n=78 in infarction group) and non-posterior circulation infarction (n=30 in TIA group).Color Doppler flow imaging and transcranial color Doppler ultrasonography were used to examine the contralateral vertebral artery extracranial diameter, peak systolic velocity(PSV) and end diastolic velocity(EDV) of bilateral extracranial and intracranial vertebral arteries.The differences of the vertebral artery occlusion types,establishment of collateral circulation and hemodynamic changes of the contralateral vertebral artery were compared between the two groups.Results The patients with single vertebral artery occlusion in the infarction group and TIA group were 69 (88.5%) and 26 (86.7%) respectively;those with bilateral vertebral artery occlusion were 9 (11.5%) and 4 (13.3%) respectively.There was no significant difference in the number of vertebral artery occlusion between the two groups (χ2=0.000,P=1.000).The proportion of patients with vertebral artery occlusion in intracranial segment in the infarction group was higher than that in the TIA group (70.5% [55/78] vs.36.7% (11/30);χ2=10.444,P=0.001).The proportion of patients with the establishment of collateral circulation in the infarction group was lower than that in the TIA group (14.1% [11/78] vs.43.3% (13/30);χ2=10.711,P=0.001).The peak systolic velocity (PSV) and the end diastolic velocity (EDV) of contralateral extracranial vertebral artery in patients with single vertebral artery occlusion in the TIA group were higher than those in the infarction group (65±21 cm/s vs.57±15 cm/s,25±8 cm/s vs.20±7 cm/s,t=2.043 and 2.606 respectively,all P<0.05).Conclusion The establishment of collateral circulation and hemodynamic compensation of the contralateral vertebral artery after vertebral artery occlusion were closely associated with the occurrence of posterior circulation ischemia.

Objective To study the value of score for neonatal acute physiology Ⅱ(SNAP]Ⅱ) and its extension version Ⅱ (SNAPPE-Ⅱ) in predicting neonatal necrotizing enterocolitis (NEC) outcome.Methods We explored 73 NEC patients by statistics who were treated in our hospital from January 2002 to January 2012.The patients were divided into two groups:surgery group and non-surgery group,then they were divided into subgroups:alive group and death group.The general information including birth weight,age,clinical manifestations,treatment of patients were collected.Every patient was checked and scored by the methods SNAP-Ⅱ] and SNAPPE-Ⅱ in time.Results The scores (27.0 ± 2.3,26.5 ± 1.8) of surgery group including SNAP-Ⅱ and SNAPPE-Ⅱ were higher than those (14.0 ± 2.1,15.0 ± 2.5) in the non-surgery group(P ＜ 0.01).The scores(31.0 ± 3.2,31.0 ± 3.4) of the death group including SNAP-Ⅱ and SNAPPE-Ⅱ were higher than those(11.0 ± 2.5,10.0 ± 3.6) in the alive group(P ＜ 0.01).According to the area under the curve(AUC) analyzed by the receiver operating characteristic(ROC) curve for measuring the scores of surgery predicting,AUC was 0.726 for SNAP-Ⅱ and 0.732 for SNAPPE-Ⅱ.The value of predicting surgery risk was 20 and 24 respectively.According to the AUC analyzed by the ROC curve for measuring the scores for surgery predicting,AUC was 0.752 for SNAP-Ⅱ and 0.825 for SNAPPE-Ⅱ.The value of predicting mortality risk was 31 and 33 respectively.All P values were less than 0.01 and there were significant differences.Conclusion The two kinds of score for neonatal acute physiology have an important significance in predicting surgery and mortality risk of NEC.

Objective To study the relationship between umbilical cord leptin levels and fetal growth as well as neonatal birth weight. Methods One hundred and forty - two neonates selected from February 2009 to June 2013 in Shangqiu First People's Hospital according to the different gestational age and birth weight were divided into 3 groups. Group A included 44 cases(small for gestational age,birth weight below the average weight of the 10th percentile at the same gestational age),23 boy cases,21 girl cases;group B included 56 cases(appropriate for gestational age,birth weight at the average weight of the 10th to 90th percentile at the same gestational age),30 boy cases,26 girl cases;group C included 42 cases(large for gestational age,birth weight above the average weight of the 90th percentile at the same gestational age),22 boy cases,20 girl cases. Neonatal body mass index,birth weight,placenta weight and umbilical lep-tin levels of three groups were compared. Results Neonatal birth weight,neonatal body length,body mass index and the placenta weight leptin levels in group A were significantly lower than those of group B,having statistically significant difference(all P ﹤ 0. 001);Neonatal birth weight,neonatal body length,body mass index and the placenta weight leptin levels in group C were significantly higher than those in group B,with statistically significant difference( all P ﹤0. 001). Neonatal birth weight in the boy group was obviously higher than that of the girl group,and the difference was statistically significant(P ﹤ 0. 001). Neonatal leptin levels in the boy group were significantly lower than that of the girl group,and the difference was statistically significant(P ﹤ 0. 001). There were positive correlations between the umbili-cal cord leptin levels and the neonatal birth weight,neonatal length,neonatal weight index and the placenta weight(r =0. 382,0. 276,0. 358,0. 412,all P ﹤ 0. 01). Conclusions The umbilical cord leptin levels are closely associated with neonatal birth weight and intrauterine growth retardation,and it can be used as one of the important indicators for reflec-ting neonatal birth weight and fetal growth.

Objective To evaluate the short-term and long-term effects in patients of carotid artery subtotal or complete occlusion after carotid endarterectomy ( CEA) using vascular ultrasound. Methods A total of 107 consecutive patients were diagnosed as carotid artery occlusive disease with DSA and treated with CEA at Beijing Xuanwu Hospital,Capital Medical University from January 2005 to January 2014 were enrolled retrospectively. Sixty-three of them had subtotal occlusion ( the carotid artery stenosis rate 95% to 99%) and 44 had complete occlusion. The occurrence of perioperative complications of all patients was documented. The follow-up study used outpatient follow-up and telephone tracking. The patients of surgical recanalization were followed up with ultrasound at 1 week, 3, 6, 12, and 24 months after procedure. The clinical prognosis, restenosis, vascular structure and hemodynamic changes of the patients after CEA were documented. Results (1) Of the 107 patients,86 (80. 4%) achieved recanalization after procedure and 21 (19. 6%) did not. The incidence of stroke and death was 4. 7% (5 cases) within 30 days after procedure,among them the incidence of subtotal occlusion group was 4. 8% ( n=3 cases) and the complete occlusion group was 4. 5% (2 cases). (2) Within one week after procedure,the peak systolic velocity ( PSV) ,end diastolic velocity ( EDV) ,and pulsatility index ( PI) of the ipsilateral middle cerebral artery in the recanalization patients increased significantly (120 ± 39 cm/s vs 60 ± 17 cm/s,50 ± 18 cm/s vs 33±11cm/s,and0.96±0.20vs0.67±0.14,respectively).Thereweresignificantdifferences(allP<0. 01). Carotid artery ultrasound showed that the local vessel diameters of the original lesions in the recanalization patients were widened as compared with preoperation (4. 4 ± 1. 1 and 3. 6 ± 1. 0 mm). There was significant difference (P<0. 01). (3) Sixty-nine patients with recanalization were followed up for 1 to 60 months( the median time was 12 months) . One to six months after procedure,the patency rate of the patients was 95. 6%(n=66),>6 to 12 months was 94. 2%(n=65),>12 to 24 months was 94. 2%(n=65),and more than 2 years was 91. 3%(n=63). Conclusion Vascular ultrasound can conduct short-term and long-term follow-up for carotid artery occlusive disease after CEA. The degree of blood flow improvement should be identified and restenosis should be found in time after procedure.