随着铜绿假单胞菌（铜绿）的耐药性逐年增强，铜绿感染已经成为公共医疗卫生的重点关注问题。线粒体自噬及其介导的线粒体功能障碍在多种细菌感染中已被报道，但线粒体功能障碍在宿主调控铜绿感染中的作用尚不明确。因此，本研究建立铜绿刺激小鼠巨噬细胞感染模型和小鼠急性铜绿感染模型，探讨铜绿是否通过诱导线粒体自噬改变线粒体功能，进而影响宿主免疫炎症反应和细胞毒性，并通过监测生存率和肺组织病理学变化进一步确定线粒体自噬在小鼠铜绿体内感染模型中的作用。结果表明，铜绿引起小鼠腹腔巨噬细胞线粒体功能障碍，并通过线粒体自噬途径清除铜绿刺激引起的活性氧（ROS）累积，从而抑制铜绿引起的促炎性细胞因子分泌并增强细胞毒性。体内实验进一步确认线粒体自噬在铜绿体内感染中的作用。
Mice ; Animals ; Reactive Oxygen Species/metabolism* ; Pseudomonas aeruginosa ; Macrophages/metabolism* ; Mitochondria ; Cytokines/metabolism*

[摘 要] 目的：探讨抑制素β亚基A反义RNA1（INHBA-AS1）对宫颈癌HeLa细胞EMT和鸟氨酸代谢途径的影响及其机制。方法：体外常规培养HeLa细胞，实验分为10组：对照组、阴性对照（NC）组、sh-INHBA-AS1组、PluriSIn 1[硬脂酰辅酶A去饱和酶（stearyl CoA desaturase，SCD）抑制剂]组、NC+PluriSIn 1组、sh-INHBA-AS1+PluriSIn 1组、10058-F4（c-Myc抑制剂）组、NC+10058-F4组、sh-INHBA-AS1+10058-F4组、sh-INHBA-AS1+OE-c-Myc组。平板克隆实验检测各组细胞的增殖能力，FCM检测各组细胞的凋亡情况，Transwell小室实验检测各组细胞的侵袭、迁移能力，qPCR法检测各组细胞中INHBA-AS1、c-Myc、SCD和EMT相关基因（N-cadherin、TGF-β、ZEB1）mRNA的表达，WB法检测各组细胞中c-Myc、SCD、EMT相关（N-cadherin、TGF-β、ZEB1）、S-腺苷-甲硫氨酸脱羧酶（SAMDC）和亚精胺/精胺N1-乙酰转移酶（SSAT）蛋白的表达，ELISA检测各组细胞上清液中鸟氨酸脱羧酶（ODC）的含量。结果：敲减INHBA-AS1表达使HeLa细胞的增殖、侵袭和迁移能力显著降低（均P<0.05）而细胞凋亡率显著升高（P<0.05），qPCR、WB法检测结果显示，敲减INHBA-AS1均可显著抑制HeLa细胞中c-Myc、SCD、N-cadherin、TGF-β、ZEB1和SAMDC的表达（均P<0.05），而促进SSAT的表达（P<0.05），并降低HeLa细胞上清液中ODC的含量（P<0.05）。与c-Myc抑制剂和SCD抑制剂单独处理相比，其联合敲减INHBA-AS1后上述作用更加显著（均P<0.05）；与sh-INHBA-AS1组相比，进一步过表达c-Myc后HeLa细胞的增殖能力显著升高（P<0.05）、SCD和N-cadherin蛋白表达水平显著升高（P<0.05）、细胞上清液中ODC含量显著升高（P<0.05）。结论：INHBA-AS1可通过c-Myc调控SCD的表达，从而影响HeLa细胞鸟氨酸代谢和EMT进程，进而促进HeLa细胞的增殖、侵袭和迁移能力。

Single-cell or low-input multi-omics techniques have revolutionized the study of pre-implantation embryo development.However,the single-cell or low-input proteomic research in this field is rela-tively underdeveloped because of the higher threshold of the starting material for mammalian embryo samples and the lack of hypersensitive proteome technology.In this study,a comprehensive solution of ultrasensitive proteome technology(CS-UPT)was developed for single-cell or low-input mouse oocyte/embryo samples.The deep coverage and high-throughput routes significantly reduced the starting material and were selected by investigators based on their demands.Using the deep coverage route,we provided the first large-scale snapshot of the very early stage of mouse maternal-to-zygotic transition,including almost 5,500 protein groups from 20 mouse oocytes or zygotes for each sample.Moreover,significant protein regulatory networks centered on transcription factors and kinases between the MII oocyte and 1-cell embryo provided rich insights into minor zygotic genome activation.

Objective:To compare the restoration effects and mechanical reconstruction between different approaches in percutaneous kyphoplasty (PKP) through an in vitro mechanical experiment. Methods:T 7 to L 4 segments of adult male embalmed spinal specimens were selected for this experiment. Single vertebral specimens were randomly divided into 4 groups: unilateral angled approach group (Group A), unilateral transpedicular approach group (Group B), unilateral oblique approach group (Group C), and bilateral transpedicular approach group (Group D) ( n=10). The anterior and posterior edges of the vertebral body were measured, and the vertebral volumes were calculated and compared. After the model of osteoporotic vertebral compression fracture (OVCF) was established on a biomechanical machine, the anterior and posterior edges of the vertebral body were measured again. After the 4 groups of specimens were subjected to PKP via different approaches, Micro-CT examination of the vertebral bodies was conducted to measure the postoperative anterior and posterior edges of the vertebral body. The original strength and stiffness of the vertebral body, the stiffness after modeling, the postoperative strength, the postoperative stiffness on the puncture and contralateral sides, and postoperative overall stiffness were recorded. The distribution of bone cement in the vertebral body, recovery of anterior and posterior heights, strength, and stiffness were compared among the 4 groups. Results:There was no statistically significant difference in the vertebral volume among the 4 groups ( P>0.05). The amount of bone cement in group D was significantly larger than that in the other 3 groups ( P<0.05). There was no statistically significant difference among the 4 groups in terms of vertebral height recovery, original strength, original stiffness, stiffness after modeling, or postoperative overall stiffness ( P>0.05). There was no statistically significant difference between the postoperative strength and the original strength in the 4 groups ( P>0.05). The postoperative stiffness on the puncture side in the 4 groups and the postoperative stiffness on the contralateral side in groups A and D were significantly higher than those after modeling ( P<0.05), but there was no statistically significant difference in the contralateral stiffness in groups B and C between postoperation and post-modeling ( P>0.05). Conclusions:In PKP, the unilateral angled approach, unilateral transpedicular approach, unilateral oblique approach, and bilateral transpedicular approach all can effectively restore the height, strength and overall stiffness of the responsible vertebral body. The unilateral angled approach and the bilateral transpedicular approach can achieve balanced restoration of the stiffness on bilateral sides of the responsible vertebral body.

Alcoholic liver disease(ALD)is the most frequent liver disease worldwide,resulting in severe harm to personal health and posing a serious burden to public health.Based on the reported antioxidant and anti-inflammatory capacities of scutellarin(SCU),this study investigated its protective role in male BALB/c mice with acute alcoholic liver injury after oral administration(10,25,and 50 mg/kg).The results indicated that SCU could lessen serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels and improve the histopathological changes in acute alcoholic liver;it reduced alcohol-induced malondialdehyde(MDA)content and increased glutathione peroxidase(GSH-Px),catalase(CAT),and superoxide dismutase(SOD)activity.Furthermore,SCU decreased tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β messenger RNA(mRNA)expression levels,weakened inducible nitric oxide synthase(iNOS)activity,and inhibited nucleotide-binding oligomerization domain(NOD)-like receptor protein 3(NLRP3)inflammasome activation.Mechanistically,SCU suppressed cytochrome P450 family 2 subfamily E member 1(CYP2E1)upregulation triggered by alcohol,increased the expression of oxidative stress-related nuclear factor erythroid 2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)pathways,and suppressed the inflammation-related degradation of inhibitor of nuclear factor-κB(NF-κB)-α(IκBα)as well as activation of NF-κB by mediating the protein kinase B(AKT)and p38 mitogen-activated protein kinase(MAPK)pathways.These findings demonstrate that SCU protects against acute alcoholic liver injury via inhibiting oxidative stress by regulating the Nrf2/HO-1 pathway and suppressing inflammation by regulating the AKT,p38 MAPK/NF-κB pathways.

Objective:To investigate the effect of the artificial facial nerve on the restoration of orbicularis oculi muscle function for unilateral peripheral facial paralysis in rabbit.Methods:Artificial facial nerve was implanted into the rabbit with unilateral peripheral facial paralysis between January 2018 and May 2019. At different time points after operation, the affected orbicularis oculi muscles′ stimulation closure threshold and the synchronism about the motion of the two sides of orbicularis oculi muscles were monitored. T test was used with SPSS 13.0 software. Results:There was no significant difference in the closure threshold of the orbicularis oculi muscle on the 7th and 28th days after artificial facial nerve implantation ( P>0.05). The synchronism of the normal side eye closing movement triggered the affected orbicular oculi muscle movement with the artificial facial nerve was up 19/20 to 20/20 at different times. Conclusions:The artificial facial nerve system had the features of stable working condition and high synchronizing effect for stimulating movement. It could restore the closed eye function in animals with peripheral facial paralysis animals and had great clinical application prospects.

Objective To analyze the multi-component,multi-target and multi-pathway mechanism of peony leaf in the treatment of cervical cancer based on network pharmacology. Methods The possible active components and targets in peony leaf were screened and predicted by pharmacological database and analysis platform of traditional Chinese medicine system,and the related targets of cervical cancer diseases were ob-tained by searching Therapeutic Target Database and others. The potential targets for the disease regulation were screened according to the active components of peony leaves entering blood,then the key target names and the pathways involved in the treatment of peony leaf were selected according to the network topological characteristic parameters. Then,the enrichment analysis was carried out by using ClueGO software plug-in. Results There were 194 target sites for 11 blood entry components in peony leaves. Finally,171 signal pathways were ob-tained,and 21 key pathways related to cervical cancer were obtained after the wide pathway was excluded,such as estrogen signaling pathway,neurotrophin signaling pathway and so on. Conclusion Peony leaves may play a vital role in the treatment of cervical cancer by acting on inflammatory,metabolic,immunological,endocrine and cell cycle related protein targets and pathways.

Objective:To establish the fingerprint analysis method for the root of Rosa laevigata Michx from different regions by UPLC. Methods:The column was ACQUITY UPLC? Phenyl(2.1 mm × 100 mm,1.7 μm). The mobile phase consisted of methanol-water with gradient elution. The flow rate was 0. 2 ml·min-1 , the detection wavelength was 210 nm, the column temperature was 30℃, and the injection volume was 3 μl. Results:The fingerprint consisted of 15 common peaks. The range of similarity for twelve bat-ches of the root of R. laevigata Michx was 0. 489-0. 942. And the reference fingerprint of the root of R. laevigata Michx was estab-lished by UPLC. Conclusion:The fingerprint method is simple and reproducible, which can provide basis for the quality control and the medicinal resources exploration.

Objective To investigate the impact of early initiation of continuous renal replacement therapy (CRRT) based on Kidney Disease: Improving Global Outcomes (KDIGO) classification on the prognosis of critically ill patients with acute kidney injury (AKI). Methods A retrospective analysis of clinical data of patients diagnosed as AKI in Department of Critical Care Medicine of Zhejiang Provincial People's Hospital from January 2011 to January 2015 was conducted. All patients included should be 18 years old or older, having stayed in intensive care unit (ICU) for more than 48 hours, and received CRRT. All subjects were divided into three groups according to their renal function before CRRT according to the KDIGO-AKI guideline: AKI-stage 1 group, AKI-stage 2 group and AKI-stage 3 group. The general condition, original disease, severity of disease, duration of mechanical ventilation, the length of ICU or hospital stay, 28-day survival rate and in-hospital mortality rate were compared among these three groups. Additionally, risk factors for the 28-day survival rate and hospital mortality of critically ill patients with AKI were screened by logistic regression analysis. Results A total of 258 critically ill patients with AKI were enrolled, with 64 cases in AKI-stage 1 group, 62 cases in AKI-stage 2 group, and 132 cases in AKI-stage 3 group. 116 patients survived with 28-day survival rate of 44.96%. 154 patients died with hospital mortality 59.69%. The precipitating factors of AKI in all three groups (stage 1, stage 2, and stage 3) were similar, with sepsis, heart failure and poisoning (drugs or poison) being the main triggers for AKI, accounting for 35.66%, 19.38% and 13.18%, respectively. There were significant differences in the rate of vasoactive agent usage (31.25%, 41.94%, 50.00%, χ2 = 6.241, P = 0.044), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score (20.87±7.37, 17.19±7.02, 22.58±7.95, F = 5.292, P = 0.006) and sequential organ failure assessment (SOFA) score (8.41±3.46, 6.22±2.43, 9.58±3.71, F = 10.328, P = 0.000), while there was no significant difference in gender, age, primary disease, time from ICU admission to the beginning of CRRT, mean arterial pressure (MAP), lactate level or 24-hour lactate clearance rate (LCR), mechanical ventilation time, the length of ICU or hospital stay, 28-day survival rate or hospital mortality among these three groups (all P > 0.05). According to the logistic regression analysis, time from ICU admission to start of CRRT and lactate level were the independent risk factors for 28-day survival rate or hospital mortality of critically ill patients with AKI [odds ratio (OR) for 28-day survival rate was 0.850 and 0.774, 95% confidence interval (95%CI) was 0.752-0.960 and 0.638-0.940, P value was 0.009 and 0.010, respectively; OR for hospital mortality was 0.884 and 0.756, 95%CI was 0.781-1.000 and 0.610-0.939, P value was 0.049 and 0.011, respectively]. Conclusion Early initiation of CRRT based on KDIGO-AKI classification could not improve the prognosis of critically ill patients with AKI, the optimal timing of RRT for such patients remains to be further explored.

Purpose To investigate the difference of imaging features of tree-in-bud (TIB) sign at CT between stage Ⅰ central lung cancer and bronchial disseminated pulmonary tuberculosis in order to reduce the misdiagnosis rate of central lung cancer.Materials and Methods 32 patients of stage Ⅰ central type lung cancer confirmed pathologically or clinically (lung cancer group) and 47 patients of bronchial disseminated pulmonary tuberculosis confirmed pathologically or clinically (tuberculosis group) underwent chest CT scanning and were found with TIB sign.The imaging data of all the cases were analyzed retrospectively in terms of distribution range,accompanying CT findings of TIB sign,and abnormalities of segmental and (or) larger bronchus proximal to it.Results TIB sign was mainly focal (unilobar) in lung cancer group (96.88％),while it was mainly diffuse (more than one pulmonary lobe) in tuberculosis group (80.85％).The difference was significant (P＜0.01).Accompanying CT findings:① Obstructive bronchial mucoid impaction distal to the TIB was more common in lung cancer group than in tuberculosis group (100％ vs 6.38％,P＜0.01).② Consolidation and/or ground-glass opacities were found both in lung cancer group (34.38％) and in tuberculosis group (48.94％).The difference was not significant (P＞0.05).③ Cavity was less common in lung cancer group than in tuberculosis group (0 vs 38.30％,P＜0.01).④ Nodules with diameter larger than 5 mm were much fewer in lung cancer group than in tuberculosis group (0 vs 76.60％,P＜0.01).TIB sign was found in 33 lobes in lung cancer group,and the abnormity was found in all these lobes (100％);while in tuberculosis group,TIB sign was found in 144 lobes and the abnormity was found in 56 lobes (38.89％).Conclusion TIB sign shows differences between stage Ⅰ central lung cancer and bronchial disseminated pulmonary tuberculosis.The TIB signs such as focal distribution,accompanied by bronchial mucoid impaction and abnormality of segmental or (and) larger bronchus may predict the high possibility of central lung cancer.