Carcinoma-associated fibroblasts (CAFs) are the main cellular components of the tumor microenvironment and promote cancer progression by modifying the extracellular matrix (ECM). The tumor-associated ECM is characterized by collagen crosslinking catalyzed by lysyl oxidase (LOX). Small extracellular vesicles (sEVs) mediate cell-cell communication. However, the interactions between sEVs and the ECM remain unclear. Here, we demonstrated that sEVs released from oral squamous cell carcinoma (OSCC)-derived CAFs induce collagen crosslinking, thereby promoting epithelial-mesenchymal transition (EMT). CAF sEVs preferably bound to the ECM rather than being taken up by fibroblasts and induced collagen crosslinking, and a LOX inhibitor or blocking antibody suppressed this effect. Active LOX (αLOX), but not the LOX precursor, was enriched in CAF sEVs and interacted with periostin, fibronectin, and bone morphogenetic protein-1 on the surface of sEVs. CAF sEV-associated integrin α2β1 mediated the binding of CAF sEVs to collagen I, and blocking integrin α2β1 inhibited collagen crosslinking by interfering with CAF sEV binding to collagen I. CAF sEV-induced collagen crosslinking promoted the EMT of OSCC through FAK/paxillin/YAP pathway. Taken together, these findings reveal a novel role of CAF sEVs in tumor ECM remodeling, suggesting a critical mechanism for CAF-induced EMT of cancer cells.
Humans ; Paxillin/metabolism* ; Protein-Lysine 6-Oxidase/metabolism* ; Carcinoma, Squamous Cell/pathology* ; Epithelial-Mesenchymal Transition ; Integrin alpha2beta1/metabolism* ; Mouth Neoplasms/pathology* ; Collagen/metabolism* ; Fibroblasts ; Extracellular Vesicles/metabolism* ; Cell Line, Tumor ; Tumor Microenvironment

【Objective】 To investigate the protection of astragaloside IV from high glucose induced podocyte injury and mitochondrial dysfunction and its molecular mechanisms. 【Methods】 The model of podocyte injury induced by high glucose (30 mmol/L glucose) was established, and the model cells were treated with low, medium and high doses of astragaloside IV respectively; cell activity was detected by CCK-8. Apoptosis was detected by TUNEL staining. Mitochondrial membrane potential was detected by JC-1 fluorescence probe. ATP content was detected by the kit. The expression levels of apoptosis and podocyte injury related proteins and Notch pathway related proteins were detected by Western blotting. 【Results】 Compared with the control group, cell activity was decreased, apoptosis level was increased (P<0.05), anti-apoptotic protein (Bcl2) expression was decreased, and apoptosis protein (Bax, cleaved-caspase 9, cleaved-caspase 3) expressions were increased (all P<0.05) in HG group. Compared with HG group, HG+AS-IV improved cell activity and apoptosis level induced by high glucose (P<0.05), increased expression of anti-apoptotic protein (Bcl2), and decreased expressions of apoptotic protein (Bax, cleaved-caspase 9, and cleaved-caspase 3) (all P<0.05). Compared with the control group, mitochondrial dysfunction occurred in HG group, JC-1 monomer content increased, and ATP content decreased (all P<0.05). Compared with HG group, HG+AS-IV improved mitochondrial dysfunction, increased JC-1 polymer content and ATP content (P<0.05). In addition, compared with the control group, the expression of Notch pathway-related protein was decreased in HG group (P<0.05). Compared with HG group, Notch pathway-related protein expression was increased in HG+AS-IV group (all P<0.05). Molecular docking results showed that AS-IV could bind Notch1. 【Conclusion】 Astragaloside IV can improve podocyte injury and mitochondrial dysfunction induced by high glucose, possibly by inhibiting Notch pathway activation.

Objective:To investigate the effect of motion correction (MC) on the calculated values of myocardial blood flow (MBF) and myocardial flow reserve (MFR) based on cadmium-zinc-telluride SPECT (CZT SPECT) images.Methods:Twenty-eight consecutive patients (10 males, 18 females, age: (60.75±11.62) years) with suspected or known coronary artery disease who underwent myocardial perfusion imaging (MPI) with dynamic CZT SPECT between June 2019 and August 2019 in TEDA International Cardiovascular Hospital were retrospectively analyzed. The MBF and MFR during rest imaging and stress imaging were quantitatively analyzed. Corridor 4DM software was used to calculate the stress MBF (sMBF) and MFR of the coronary artery branches and left ventricular (LV) before and after MC. The paired t test and Pearson correlation were used for data analysis. Results:The sMBF and MFR of LV before MC were (0.82±0.49) ml·min -1·g -1 and 1.69±0.68 respectively. After MC the two parameters increased to (1.05±0.64) ml·min -1·g -1 and 2.12±0.77 respectively ( t values: -4.87, -6.01, both P<0.001). The sMBF and MFR in left anterior descending (LAD), left circumflex (LCX), right coronary artery (RCA) and LV before MC were correlated with those after MC ( r values: 0.69-0.96, all P<0.001). If MFR <2.0 was used as the reference of impaired MFR, data before MC showed 19 patients (67.9%, 19/28) had impaired MFR, while 13 patients (46.4%, 13/28) had impaired MFR based on MFR values after MC. Conclusion:For MPI quantitative imaging with CZT SPECT, the calculated values of sMBF and MFR after MC is higher than those before MC, suggesting that MC is helpful to reduce the false positive results which may be caused by the " creep" effect of the heart.

【Objective】 To explore the main development directions and research progress of blood transfusion in the U. S.military based on ClinicalTrials.gov. 【Methods】 The literature of clinical trials related to field transfusion medicine from 2002 to 2021 were retrieved based on www.clinicaltrials.gov with "United States Department of Defense" , "U.S.Army Medical Research and Development Command" and "United States Naval Medical Research Center" as the search terms, and the main characteristics were analyzed in terms of research progress, clinical stage, task allocation, regional distribution, research contents and results. 【Results】 A total of 931 clinical trials were retrieved, of which 16 were closely related to field blood transfusion medicine.Among them, pathogen reduction/inactivation of whole blood, platelet transfusion, plasma transfusion, whole blood transfusion and other types of researches accounted for 25% (4/16), 25% (4/16), 25% (4/16), 18.75 (3/16) and 6.25% (1/16), respectively.The U. S.military focused on the new technological breakthroughs in blood safety and blood storage, the evaluation of the countershock therapy, the application of whole blood early transfusion and the civil-military inosculation in research. 【Conclusion】 This study, based on ClinicalTrials.gov data, redounds to understand the research progress and current situation of the U. S.military field blood transfusion medicine.

Objective:To describe the COVID-19 epidemic and its characteristics in Heilongjiang province, and provide evidence for the further prevention and control of COVID-19 in the province.Methods:The information of COVID-19 cases and clusters were collected from national notifiable disease report system and management information system for reporting public health emergencies of China CDC. The Software’s of Excel 2010 and SPSS 23.0 were applied for data cleaning and statistical analysis on the population, time and area distributions of COVID-19 cases.Results:On January 22, 2020, the first confirmed case of COVID-19 was reported in Heilongjiang. By March 11, 2020, a total of 482 cases domestic case of COVID-19, The incidence rate was 1.28/100 000, the mortality rate was 2.70% (13/482) in 13 municipalities in Heilongjiang. There were 81 clusters of COVID-19, The number of confirmed cases accounted for 79.25% (382/482) of the total confirmed cases and 12 cases of deaths. The family clusters accounted for 86.42% (70/81). Compared with the sporadic cases, the mortality rate, proportion of elderly cases aged 60 or above and severe or critical cases of clinical classification were all higher in the clusters especially the family clusters, but the differences were not significant ( P>0.05). There were 34 clusters involving more than 5 confirmed cases accounted for 41.98% (34/81) of the total clusters, the involved cases accounted for 68.31% (261/382) of the total cases of clusters. There were significant differences in age distribution of the cases among the case clusters with different case numbers. In the clusters involving 6-9 cases, the proportion of cases aged 65 years or above was more (26.53%, 39/147). Conclusions:The incidence rate of COVID-19 was relatively high and the early epidemic was serious in Heilongjiang, The number of cases was large in clusters especially family clusters.

Objective To apply the concept of evidence-based nursing to the practice of non-invasive ventilation,and to introduce the method of using self-made pressure-resistant cap cotton cover and its application effect.Methods Fifty patients on non-invasive ventilation admitted to intensive care unit (ICU) of Harrison International Peace Hospital Affiliated to Hebei Medical University from September 2017 to April 2018 were enrolled as observation group.By retrieved the relevant literature at home and abroad to look for clinical evidence,the concept of evidence-based nursing care program was formulated and implemented,and self-made pressure-resistant cap cotton cover was used to prevent facial pressure sores in patients on non-invasive ventilation.Forty non-invasive ventilation patients admitted from January to August in 2017 were enrolled as the historical control group,and conventional nursing was conducted by using traditional hydrocolloid dressings to prevent pressure ulcers.The incidence of facial pressure ulcers,ocular complications,dressing replacement time and cost were compared between the two groups.Results All patients were enrolled in the final analysis.The incidence of pressure ulcers in the observation group was significantly lower than that in the control group [4.0％ (2/50) vs.22.5％ (9/40),P ＜ 0.05],and the incidences of ocular complications such as eyelid swelling [2.0％ (1/50) vs.5.0％ (2/40)],conjunctival congestion [2.0％ (1/50) vs.5.0％ (2/40)] and irritant keratitis [0％ (0/50) vs.7.5％ (3/40)] were also significantly lower than those in the control group (P ＜ 0.05).Because the pressure-resistant cap cotton cover made by ourselves was easy to be replaced at any time,the interval time of dressing change in the observation group was significantly shorter than that in the control group (minutes:1.5 ± 0.5 vs.2.0 ± 0.5,P ＜ 0.05).In addition it could be cleaned,used alternately,and the cost was low,so the dressing cost in the observation group was significantly lower than that in the control group (Yuan:30±10 vs.123±20,P ＜ 0.01).Conclusion The application of self-made pressure-resistant cap cotton cover could obviously reduce the occurrence of complications such as facial pressure sores in patients on non-invasive ventilation.

Objective To study the relationship between aquaporin 9 (AQP 9) gene and brain edema in neonatal rats of hypoxic-ischemic brain damage (HIBD) and the therapeutic mechanism of mannitol.Method Healthy and 7-day-old SD rats were randomly assigned into three groups:sham-operated group,HIBD group and mannitol group.Both HIBD and mannitol group were established on HIBD model.The mannitol group was given mannitol intraperitoneally at 0,24,48 h of HIBD.2 ml/kg of 2％ Evans blue (EB) were injected intraperitoneally before sacrifice.0,6,12,24,48 and 72 h after HIBD,the outcomes were analyzed including the brain water content,the expression of AQP 9 mRNA measured using RT-PCR and immunofluorescence staining methods,and the permeability of blood-brain barrier (BBB) measured with EB.Result In HIBD group,the brain water content was higher comparing with sham-operated group at 0 h after HIBD(P ＜ 0.05),and gradually increased over time,reaching peak at 48 h (89.3％ ± 1.9％) and then decreased.In mannitol group,brain water content started to decrease from 1 h after mannitol administration to the bottom at 12 h (86.5％ ±0.6％),then increased to peak at 72 h (87.2％ ± 1.7％),and brain water content were decreased during 0 ～ 48 h comparing with HIBD group.HIBD group's EB were higher than sham-operated group (P ＜ 0.05);Mannitol group's EB were decreased comparing with HIBD group (except 0 h,P ＜ 0.05).AQP 9 mRNA expression in the HIBD group was decreased at 0 h,and reached the bottom at 48 h (0.09 ± 0.07).Comparing with sham-operated group,it was higher in the HIBD group at0,6,72 h,and lower (P＜ 0.05) at 12,24,48 h.Higher AQP 9 mRNA expression were detected in mannitol group than HIBD group and sham-operated group at each time point (with the exception of 48 h) (P ＜ 0.05).AQP 9,which was closely related to water metabolism,were widely found in the pia mater and ependyma using immunofluorescence staining.After ischemia and hypoxia insult,an increasedecrease-increase pattern of AQP 9 expression was found.Conclusion AQP 9 is widely existed in various parts of the brain,influencing brain edema through a variety of pathways.AQP 9 also plays a role in alleviating brain edema in mannitol therapy.

Objective To explore the relationship and clinical significance of Gli1 expression in peripheral blood and cancer tissues of gastric.Method 95 gastric cancer patients were recruited as surgery group,and 40 recurrent gastric cancer patients were recruited as recurrent group.50 normal subjects were recruited as control group.Paraffin specimens of gastric cancer tissues and gastric mucosa adjacent to cancer were collected to test expression of Gli1 protein.ELISA was applied to test Gli1 in peripheral blood.Results Levels of Gli1 in peripheral blood of preoperative surgery group were higher than in control group [(9.21 ± 1.37) ng/ml vs.(0.79 ±0.18) ng/ml,t =43.193,P ＜0.001].Post-up Gli1 decreased significantly to the level of control group (t =0.722,P =0.472).Levels of Gli1 in peripheral blood of recurrent group were higher than that in preoperativesurgery group (t =14.575,P ＜ 0.001).Gli1 protein in cancer tissues was higher than that in adjacent gastric mucosa (t =33.471,P ＜0.001),and Gli1 protein in cancer tissues of surgery group was higher than that in cancer tissues of recurrent group (t =8.808,P ＜0.001).Conclusion In postoperative gastric cancer patients increased Gli1 level in peripheral blood and cancer tissues predicts recurrence.

OBJECTIVETo develop and evaluate a mouse model of nonalcoholic steatohepatitis (NASH) induced by a high-fat and high-fructose (HFHFr) diet.

METHODSSix-week-old C3H mice were randomly divided into groups for HFHFr diet experimental modeling, high fat-only (HF) diet controls, high fructose-only (HFr) diet controls, and standard chow (SC) diet controls. The standard HFHFr diet was modified so that it consisted of 76.5% standard chow, 12% lard, 1% cholesterol, 5% egg yolk powder, 5% whole milk powder, and 0.5% sodium cholate, along with 20% fructose drinking water. At the end of experimental weeks 4, 8, and 16, measurements were taken for the NASH-related parameters of body mass, serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), lipid profile, and wet liver weight (upon sacrifice). In addition, histological changes in the liver were evaluated by hematoxylin-eosin (HE) and oil red O staining. The significance of differences between groups was assessed by statistical analysis, using the

METHODSof t-test, Wilcoxon rank sum test, x2 test, F test or Fisher's test as appropriate.

RESULTSAs compared to the mice in the SC group at the corresponding time points, the mice in the HFHFr and HF groups showed significantly higher body mass and wet liver weight, as well as more extensive and robust lipid disposition in hepatic tissues as evidenced by oil red O staining. However, HE staining indicated that the HFHFr and HF groups had different degrees of macrosteatosis accompanied with intralobular inflammatory foci, with the former showing more remarkable NASH-related histological changes. Analysis at the end of week 16 showed that about 80% of the mice in the HFHFr group had developed NASH [nonalcoholic fatty liver disease (NAFLD) activity score (NAS): less than 5]. The levels of low-and high-density lipoprotein (LDL and HDL) cholesterol, as well as the levels of ALT and AST, were increased from the end of week 4 to the end of week 8 for the HFHFr and HF groups. At the end of week 16, the two groups differed in the extent of increase in total cholesterol and LDL and HDL cholesterol, with only the HFHFr group showing statistically significant changes. Specifically, at the end of week 16, the HFHFr group showed ALT levels of 108.5 +/- 93.34 U/L (F=5.099, P =0.005 vs. HF group: 44.30 +/- 35.71 U/L, HFr group: 46.70 +/- 17.95 U/L, SC group: 24.70 +/- 6.57 U/L), AST levels of 316.30 +/- 208.98 U/L (F=6.654, P=0.001 vs. HF: 132.12 +/- 75.43 U/L, HFr: 143.30 +/- 38.53 U/L, SC: 122.60 +/- 12.76 U/L), total cholesterol levels of 5.18 +/- 0.58 mmol/L (F=72: 470, P =0.000 vs. HF: 3.94 +/- 0.75 mmol/L, HFr: 2.30 +/- 0.50 mmol/L, SC: 2.02 +/- 0.24 mmol/L), HDL cholesterol levels of 3.05 +/- 0.49 mmol/L (F=25.413, P =0.000 vs. HF: 2.65 +/- 0.54 mmol/L HFr: 1.77 +/- 0.47 mmol/L, SC: 1.58 +/- 0.16 mmol/L), LDL cholesterol levels of 1.11 +/- 0.23 mmol/L (F =83.297, P =0.000 vs. HF: 0.72 +/- 0.17 mmol/L, HFr: 0.27 +/- 0.04 mmol/L, SC: 0.20 +/- 0.05 mmol/ L).

CONCLUSIONThe present study suggests that a mouse model of NASH can be successfully induced by a 16-week modified HFHFr diet.

Objective To establish a mini-pool nucleic acid testing (NAT) assay using multiplex RT-nested PCR for the detection of HIV RNA, and apply it in screening for acute HIV infection among MSM. Methods Frozen EDTA plasma samples collected between Oct. 2008 and Mar. 2009 from 3 HIV infectors during window-period, a total of 30 HIV chronically infected individuals and 97 healthy subjects were used to develop the NAT assay. Plasma samples from 10 cases were pooled into one tube and centrifuged at high speed for the collection of viruses. HIV RNA was extracted. Two pairs of primers were designed according to two conserved regions of HIV RNA ( HXB2 nt 5783-nt 6228 and nt 1235-nt 2012).Multiplex RT-PCR and nested PCR were performed. Individual NAT-reactive samples were confirmed by commercially available NAT assays. The sensitivity and performance efficacy were also evaluated. The assay was then applied to 1 005 plasma specimens from MSM with negative or uncertain HIV antibody test results.These were collected in the same period as the other samples. Results ( 1 ) Two fragments of HIV were amplified successfully with the low detection limit of 162 copies/ml plasma; (2) Results of the mini-pool HIV NAT validation with samples from 3 HIV infectors during window-period were consistent with the expected values; (3) All 30 plasma samples from MSM with positive HIV antibody, which were tested by multiplex RT nested PCR, were found to be HIV RNA positive; (4) One out of 1 005 plasma samples was found to be HIV RNA positive, for this case acute infection was followed-up and sero-conversion was found. Conclusion Mini-pool NAT has good sensitivity, and may be applied to screening HIV RNA among MSM during window-period.