AIM: To investigate the relationship between vascular smooth muscle cell (VSMC) injury, organelle stress response and autophagic cell death (autophagy) and ferroptosis induced by the chemical hypoxia inducer cobalt chloride (CoCl2) through the bioinformatics analysis and in vitro cell experimentation. METHODS: The dataset GSE119226 of VSMC treated with cobalt chloride was acquired from the gene expression database (GEO). The R language was used to investigate the relationship between CoCl2 treatment and organelle stress response (Golgi stress, endoplasmic reticulum stress) and two forms of cell death (ferroptosis and autophagic cell death). With primary cultured rat VSMC (rVSMC) and CoCl2-induced anoxia model, the changes in cell viability were detected by CCK-8 method, and reactive oxygen species (ROS) levels were measured using DCFH-DA method. The expression levels of HIF-1α (a key molecule in hypoxia), Golgi stress markers GM130 and p115, endoplasmic reticulum stress markers GRP78 and CHOP, autophagy markers LC3-II / LC3-I and Beclin1, and ferroptosis markers GPx4 and xCT were detected by Western blot. The effect of inducing or inhibiting organelle stress and cell death on the CoCl2-induced cell damage was also observed. RESULTS: Differentially expressed genes analysis of GSE119226 dataset showed that CoCl2 treatment of VSMCs had significant effects on organelle function and stress response, autophagy and ferroptosis-related genes, in which endoplasmic reticulum stress, protein processing in endoplasmic reticulum, regulation of Golgi to plasma membrane protein transport, autophagy / autophagic cell death, and ferroptosis pathways were remarkably enriched. The results of in vitro experiment showed that compared with normal rVSMC, cell viability was significantly decreased after CoCl2 treatment, as well as HIF-1α protein expression and ROS levels in rVSMCs were increased. In rVSMC treated with Co-Cl2, the expression levels of Golgi structural proteins GM130 and p115 (reflecting the occurrence of Golgi stress) were decreased, while the markers GRP78 and CHOP (reflecting the occurrence of endoplasmic reticulum stress) were increased. At the same time, CoCl2 treatment also reduced the expression of autophagy markers LC3-II/LC3-I and Beclin1 (indicating the decrease levels of autophagy), while the expression of ferroptosis markers GPx4 and xCT were decreased (indicating the occurrence of ferroptosis). Compared with CoCl2 treatment group, induced Golgi stress, endoplasmic reticulum stress, or ferroptosis could further reduce cell viability, while inhibition of these processes could improve cell viability. On the other hand, increasing the level of autophagy can improve the cell viability. CONCLUSION: Hypoxia induced by cobalt chloride can lead to VSMC injury. Golgi stress, endoplasmic reticulum stress, ferroptosis, and the reduction of autophagy level play an important role in it. Inhibition of organelle stress response and ferroptosis, or increase of autophagy level can improve VSMC injury caused by cobalt chloride.

Objective To investigate the efficacy and safety of artificial liver support therapy with an Evanure-4A selective membrane plasma separator and its influence on platelet count in the treatment of patients with acute-on-chronic liver failure(ACLF)patients with different platelet counts.Methods A total of 302 patients with ACLF who were hospitalized in Department of Hepatology,Chengdu Public Health Clinical Medical Center,from January 2021 to May 2023,were enrolled,and according to the platelet count(PLT),they were divided into group A(25×109/L—50×109/L)with 101 patients,group B(51×109/L—80×109/L)with 98 patients,and group C(81×109/L—100×109/L)with 103 patients.In addition to medical treatment,all patients received different modes of artificial liver support therapy based on their conditions,including plasma perfusion combined with plasma exchange,double plasma molecular adsorption combined with plasma exchange,and bilirubin system adsorption combined with plasma exchange.The paired t-test was used for comparison of continuous data before and after treatment in each group;an analysis of variance was used for comparison between multiple groups,and the SNK-q test was used for further comparison between two groups;the chi-square test was used for comparison of categorical data between multiple groups.Results Of all 302 patients,268(88.74%)achieved varying degrees of improvement in clinical symptoms after artificial liver support therapy.After treatment,all three groups had varying degrees of reductions in alanine aminotransferase(t=14.755,21.614,and 15.965,all P＜0.001),aspartate aminotransferase(t=11.491,19.301,and 13.919,all P＜0.001),total bilirubin(t=19.182,17.486,and 21.75,all P＜0.001),and international normalized ratio(INR)(t=3.497,3.327,and 4.358,all P＜0.05).After artificial liver support therapy with an Evanure-4A selective membrane plasma separator,PLT in group A decreased from(37.73±6.27)×109/L before treatment to(36.59±7.96)×109/L after treatment,PLT in group B decreased from(66.97±7.64)×109/L before treatment to(62.59±7.37)×109/L after treatment,and PLT in group C decreased from(93.82±5.38)×109/L before treatment to(85.99±12.49)×109/L after treatment;groups B and C had significant reductions in PLT after treatment(t=12.993 and 8.240,both P＜0.001),but there was no significant difference in group A(P＞0.05).There was no significant difference in the incidence rate of adverse reactions during artificial liver support therapy between the three groups(P＞0.05).Conclusion Artificial liver support therapy can improve liver function and INR in patients with ACLF.The use of Evaure-4A selective membrane plasma separator during artificial liver support therapy has little influence on platelets,and it is safe in the treatment of ACLF patients with a significantly lower level of platelets.

Objective:To analyze the results of prenatal diagnosis and outcome of pregnancy for women with a high risk for fetal aneuploidies.Methods:A total of 747 cases of prenatal diagnosis by amniocentesis due to high risks by non-invasive prenatal testing (NIPT) were selected from January 2015 to March 2022 in the Drum Tower Hospital Affiliated to Nanjing University Medical School. The amniotic fluid samples were subjected to chromosomal karyotyping and/or chromosomal microarray analysis. All cases were followed up by searching the birth information or telephone calls, and the results were recorded. 2 test or F test were used for comparing the difference between the groups.Results:Among the 747 pregnant women with a high risk by NIPT, 387 were true positives, and the overall positive predictive value (PPV) was 51.81%. The PPVs for trisomy 21 (T21), trisomy 18 (T18), trisomy 13 (T13) and sex chromosome aneuploidies (SCA) were 80.24% (199/248), 60% (48/80), 14% (7/50) and 38.97% (106/272), respectively. The PPV for T21 was significantly higher than T18 and T13 ( χ2= 85.216, P<0.0001). The PPV for other chromosomal aneuploidies and copy number variations (CNVs) were 11.11% (5/45) and 40.74% (22/52), respectively. The PPV for increased X chromosomes was significantly higher than X chromosome decreases (64.29% vs. 22.22%, χ2= 5.530, P<0.05). The overall PPV for elder women (≥ 35 years old) was significantly higher than younger women (69.35% vs. 42.39%, χ2= 49.440, P<0.0001). For T21 and T18, the PPV of Z ≥ 10 group was significantly higher than that for 3 ≤ Z < 5 group or 5 ≤ Z < 10 group ( P<0.05). Among 52 cases with a high risk for CNVs, the PPV for the ≤ 5 Mb group was significantly higher than the 5 Mb < CNVs < 10 Mb or > 10 Mb groups (60% vs. 30%和60% vs. 23.53%, P<0.05). Among the 387 true positive cases, 322 had opted for induced labor, 53 had delivered with no abnormal growth and development, and 12 were lost during the follow-up. Conclusion:The PPVs for common chromosomal aneuploidies are related to the age and Z value of the pregnant women, which were higher in the elder group and higher Z value group. In addition, the PPV is associated with high risk types. The PPV for T21 was higher than T18 and T13, and that for 45, X was lower than 47, XXX, 47, XYY or 47, XXY syndrome. NIPT therefore has relatively high PPVs for the identification of chromosomal CNVs.

Objective:To explore the talent cultivation model of nursing professional group under the 1+X certificate system, to build a talent cultivation system for nursing professional groups with "three progressions, four integrations, and five cocultivation"Methods:From 1 July 2021 to 31 December 2023, built a professional group course system of "mutual sharing of foundations, mutual generation of majors, mutual selection of positions, and progressive improvement of abilities". Aligned course content with X certificate content. Deepen the reform of the three education systems, create a "dual teacher" teaching team that combines full-time and part-time work, and build teaching resources that connect the cloud and the ground. The application effect of this model was evaluated in 2021 nursing majors in Cangzhou Medical College by cluster sampling.Results:There were 748 students majoring in nursing in class of 2021, 150 boys aged 19-22(20.00 ± 1.23), 598 girls, aged 19-21(19.00 ± 2.57). There were 118 students majoring in midwifery in class of 2021, all of them were girls, aged 18-20(18.00 ± 2.11). There were 128 students majoring in rehabilitation therapy technology in class of 2021, 66 boys aged 19-21(19.00 ± 2.11), 62 girls aged 19-21(19.00 ± 2.54). Nursing professional group completed the "Basic Nursing Technology Training Guidance", "Elderly Care" and other 5 self-compiled textbooks, 730 students had participated in the X certificate assessment, 729 students e passed the assessment, the passing rate was 99.9%.Conclusions:The talent cultivation model of "three progressions, four integrations, and five co cultivation" in the nursing professional group can cultivate students′ professional abilities, and cultivate a composite technical and skilled talent with multiple abilities and interconnections in one job.

Objective To establish a model of combined injury of thermal radiation and blast wave of gas explosion in closed pipeline,and investigate the characteristics of important organ injury induced by burn-blast combined injury in rats.Methods A total of 72 male SD rats(aged 8～10 weeks,weighing 200～10 g)were randomly divided into control group and mild and severe injured groups.After the model rats were inflicted with blast wave and thermal radiation,their physical parameters were detected and measured.The respiratory function of the survival rats was tested.In 24 h later,arterial blood gas analysis,blood biochemical tests,and detection for serum inflammatory factors and lung injury related protein levels were performed,and the pathological changes in the lung tissue and trachea were observed.Results The peak range of explosive blast wave overpressure was 209～493 kPa,and the temperature was 152～258 ℃.The mortality rate was 8.3％in the mild group and 53.1％in the severe group.Compared with the control group,longer inspiratory time(Ti)and relaxation time(Tr)and larger tidal volume(TV)(P＜0.05),while lower respiratory frequency(f)were observed in the 2 injured groups within 6 h after injury(P＜0.05).At 24 h after injury,the values of partial pressure of carbon dioxide(PaCO2),residual base value(BE)and bicarbonate(HCO3-)were increased significantly(P＜0.05),while partial pressure of oxygen(PaO2)was decreased(P＜0.05).The serum levels of alanine transaminase(ALT),aspartate aminotransferase(AST),alkaline phosphatase(ALP),total bilirubin(TBI)and total bile acid(TBA)were notably elevated(P＜0.05).So were the levels of IL-1 β,TNF-α and IL-6(P＜0.01).Various severities of hyperemia foci and inflammation in the tracheal mucosa were observed in the injured rats.The incidence of lung injury was high,and the pathological manifestations were dot-shaped hemorrhage to diffuse hemorrhage of the whole lobe,rupture of alveolar septum,thickening of septum,and infiltration of red blood cells and inflammatory cells.The levels of serum lung injury-related proteins were significantly increased(P＜0.05).Conclusion A rat model which can highly simulate mild to severe burn-blast combined injury is constructed,which can meet the needs for rat model in study of explosive burn-blast compound injury.The main characteristics of the injury are lung injury,respiratory dysfunction,liver damage and secondary infection.

Objective To investigate the value of a nomogram model constructed from intratumoral and peritumoral magnetic resonance imaging radiomics combined with clinical characteristics in predicting the status of lymphovascular space invasion(LVSI)in cervical cancer.Methods A retrospective analysis was conducted on 178 cervical cancer patients confirmed by postoperative pathology,with 70 cases of LVSI(+)and 108 cases of LVSI(-).The patients were divided into a training set[142 cases,including 54 cases of LVSI(+)and 88 cases of LVSI(-)]and a test set[36 cases,including 16 cases of LVSI(+)and 20 cases of LVSI(-)]at a ratio of 8:2.All underwent magnetic resonance imaging before surgery,and regions of interest were manually delineated layer by layer on the T2WI sequence,with the peritumoral region being uniformly expanded outward.Univariate logistic analysis was performed on clinical factors to select independent factors for cervical cancer LVSI(+).Radiomic features were extracted separately from the intratumoral region,the peritumoral region,and the intratumoral-peritumoral region to construct radiomics models,and the differences between the peritumoral and the intratumoral-peritumoral models were compared.A combined model was established based on the radiomics scores of the optimal intratumoral-peritumoral model and clinical independent predictive factors,and a nomogram was plotted.Receiver operating characteristic curves were used to evaluate the diagnostic performance of each model,and decision curve analysis was used to assess the clinical value of the models.Results The combined model demonstrated the best performance among the various models,with AUC of 0.970 in the training set and 0.803 in the test set.Conclusion Intratumoral and peritumoral magnetic resonance imaging radiomics combined with clinical characteristics can effectively predict LVSI in cervical cancer.

Objective This study aimed to investigate the MRI features of forearm muscles from myotonic dystrophy type 1(DM1)and to evaluate the correlation between clinical factors and muscle MRI.Methods We retrospectively reviewed the medical records of genetic-confirmed DM1 patients including the clinical manifestations,the electromyography and the muscle pathology results.MRI scan was used to examine forearm muscles.Analysis of T1weighed images and short inversion time inversion recovery images were used to evaluate muscle fatty infiltration and muscle edema,respectively.The correlation between the fatty infiltration and age,disease duration and muscle strength were investigated.Results Twenty-three DM1 patients from different family were enrolled.The onset age was 17～60 year-old,mean age was(34.3±12.2)year-old,and the mean course of disease was 5(3,9)years.Their main symptoms were hand weakness and stiffness,accompanying with alopecia and excessive daytime sleepiness.The median fatty infiltration scores were recorded as follows:flexor digitorum profundus(FDP)4.0(3.0,4.0),flexor pollicis longus(FPL)4.0(3.0,4.0),extensor pollicis brevis 2.0(1.0,2.5),abductor pollicis longus 2.0(0.5,2.5),extensor pollicis longus 2.0(1.0,2.0),flexor digitorum superficialis 2.0(0.5,2.5),supinator(S)2.0(1.0,2.0).Other muscles scores were less than 1 of which the pronator teres(PT)was 1.0(0.0,1.0).Atrophy of FDP was detected in nine,atrophy of FPL in six and hypertrophy of PT in 11 patients.The muscle edema was evident in FDP,FPL,and the flexor digitorum superficialis,while brachioradialis and pronator teres were normal.There was no significant correlation between mean muscle fat infiltration score and age or course of disease.The mean muscle fat infiltration was significantly correlated with muscle strength score(r =-0.590,P<0.01).The mean fatty infiltration scores of FDP and FPL were not correlated with age nor course of disease,but correlated with grip strength(r =-0.754,P<0.01 and r =-0.699,P<0.01).Conclusion The forearm MRI in DM1 patients exhibit a relatively specific distribution of selective muscle involvement and can be used to assess muscle weakness.

Objective:To investigate the effects of high-energy shock and vibration on cortex injury and peripheral blood immune cells in goats.Methods:Seventeen Boer goats without gender preference were selected. By using random number tables, the goats were divided into normal control group ( n=5) and shock and vibration injury group ( n=12). The goats in the normal control group were anatomized routinely and their brain was collected after being sacrificed without any other treatment. The goats in the high-energy shock and vibration model group were placed on a loading table (part of the BY10-100 instant shock and vibration simulation platform) in a restrained state, and made into a high-energy shock and vibration injury model induced by a vertical impact waveform generator. The intravenous blood samples were taken from the goats in the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury.Then, the goats were sacrificed and the following procedures were the same as the normal control group. At 24 hours after injury, the brain injury and the histopathological changes of the cerebral cortex in the normal control group and shock and vibration injury group were observed by gross pathological and anatomical examination and HE staining. The mRNA expression of zonula occludens 1 (ZO-1), tight junction protein 5 (Claudin-5), glial fibrillary acidic protein (GFAP), ionized calcium binding adaptor molecule 1 (IBA-1), interleukin (IL)-1β, IL-6 and cluster of differentiation antigen 177 (CD177) of the cerebral cortex in the normal control group and shock and vibration injury group were measured through fluorogenic quantitative polymerase chain reaction. The expression of ZO-1 and Claudin-5 proteins of the cerebral cortex in the normal control group and shock and vibration injury group were detected by Western blotting. Hematology analyzer and coagulation analyzer were used to detect white blood cell count, neutrocyte, lymphocyte, monocyte, prothrombin time 1 (PT-1), prothrombin time international normalized ratio (PT-INR), activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin activity (PTA) and fibrinogen (FIB) levels in goats of the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury, respectively. Results:At 24 hours after injury, no visible contusion or necrosis was found in goat brain tissue in the shock and vibration injury group; the cerebral micro-vessels presented with a local dilation, hyperemia, edema, aggregation of inflammatory cells, disruption of vessel walls and leakage of red blood cells. These changes were not observed in the normal control group. In the shock and vibration injury group, ZO-1 and Claudin-5 mRNA expressions in the cerebral cortex were 0.25±0.10 and 0.09(0.04, 0.44) respectively, which were significantly lower than those of the normal control group [1.00±0.15 and 0.99(0.80, 1.20)]; GFAP, IBA-1, IL-1β, IL-6 and CD177 mRNA expression levels were 4.40(3.88, 6.75), 2.60±1.07, 3.04±0.51, 2.71±0.45 and 2.93±0.62 respectively, which were significantly higher than those of the normal control group [1.00(0.78, 1.22), 1.00±0.37, 1.00±0.27, 1.00±0.57 and 1.00±0.35]; ZO-1 and Claudin-5 protein expression levels were 0.41±0.06 and 0.42±0.11 respectively, which were significantly lower than those of the normal control group (1.08±0.12 and 0.91±0.23) (all P<0.01). In the shock and vibration injury group, the levels of white blood count, neutrocyte, and lymphocyte in peripheral blood were (13.7±3.3)×10 9/L, (35.3±14.8)% and (57.2±15.1)% respectively before injury, (19.4±3.1)×10 9/L, (60.5±12.5)% and (33.6±14.2)% respectively at 3 hours after injury, and (20.6±3.6)×10 9/L, (63.6±13.0)% and (30.9±15.0)% respectively at 6 hours after injury. By contrast, the levels of white blood count and neutrocyte were significantly increased but the level of lymphocyte was significantly decreased at 3 and 6 hours after injury ( P<0.05 or 0.01); the levels of the above indicators showed no significant changes at 0 and 24 hours after injury (all P>0.05); the level of monocyte did not change significantly at all time points before and after injury (all P>0.05). The levels of PT-1, PT-INR, APTT, TT, PTA and FIB in the shock and vibration injury group did not change significantly at each time point before and after injury (all P>0.05). Conclusion:Cerebral cortex microvascular injury and disruption of blood-brain barrier can be initiated in the early stage of high-energy shock and vibration injury in goats, accompanied by the presence of central and peripheral inflammatory response.

Objective:To evaluate the effectiveness and prognosis of emergent transcatheter aortic valve replacement (TAVR) and to provide standardized procedural suggestion for the development of emergent TAVR in China.Methods:From January 2020 to April 2021, 12 patients who underwent emergent or salvage TAVR in the Second Affiliated Hospital Zhejiang University School of Medicine were retrospectively enrolled from the TORCH registry (Transcatheter Aortic Valve Replacement Single Center Registry in Chinese Population, a prospective cohort study; NCT02803294). Baseline, periprocedural and 30-day follow up data were collected. Post-operative data were compared with pre-operative data using Paired-Samples test.Results:Patients’ median Society of Thoracic Surgeons score (STS score) was 15.432%. There was a significant decrease of mean gradient after emergent TAVR procedure (1.69 m/s vs. 4.90 m/s, P<0.01). During the 30-day follow up, there were 1 patient (8.3%) died and 2 patients received permanent pacemaker implantation. No disabling stroke, acute kidney injury, major vascular complication occurred during the first month after emergent TAVR. Among the survival patients, there was a significant releasing of heart failure symptoms to New York Heart Association function stage Ⅰ/Ⅱ in 81.8% patients at 30-day follow up. Left ventricular ejection fraction also improved significantly from (47.4±9.5)% to 58.8±8.0% ( P= 0.026). The mean gradient were (1.57±0.30) cm 2 and no patients had a moderate or severe paravalvular leakage. Besides, a significant decrease of pro-B-type natriuretic peptide (1 089.9 pg/mL vs. 12 215.5 pg/mL , P=0.001) and troponin T (0.020 ng/mL vs. 0.337 ng/mL, P=0.003) were found at 30 days after emergent TAVR. Conclusions:For patients with severe aortic stenosis and acute cardiac decompensated, emergent TAVR is a safe and effective rescue treatment.

The use of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) is the first line treatment for EGFR-mutant advanced non-small cell lung cancer (NSCLC), but drug resistance will be acquired within 1-2 years, and the following treatment efficacy is poor. The invention of programmed cell death 1 (PD-1)/programmed cell death ligand 1 (PD-L1) inhibitors has dramatically changed the situation of tumor treatment. PD-1/PD-L1 inhibitors are less effective in patients with NSCLC harboring EGFR mutation. It is a challenge to make patients with EGFR-mutated advanced NSCLC benefit from anti-PD-1/PD-L1 therapy. In this paper, the research progress on the impact of EGFR mutation on the immune status of NSCLC and related clinical studies in recent 5 years are reviewed.
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Humans ; Carcinoma, Non-Small-Cell Lung/pathology* ; B7-H1 Antigen/metabolism* ; Lung Neoplasms/pathology* ; Immune Checkpoint Inhibitors ; Ligands ; Mutation ; ErbB Receptors/metabolism* ; Protein Kinase Inhibitors/therapeutic use*