Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications.

OBJECTIVES: In the current study, for the first time, we reported a novel HCV molecular diagnostic approach termed reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticles-based lateral flow biosensor (RT-LAMP-AuNPs-LFB), which we developed for rapid, sensitive, specific, simple, and visual identification of HCV. METHODS: A set of LAMP primer was designed according to 5'untranslated region (5'UTR) gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a, which are prevalent in China. The HCV-RT-LAMP-AuNPs-LFB assay conditions, including HCV-RT-LAMP reaction temperature and time were optimized. The sensitivity, specificity, and selectivity of our assay were evaluated in the current study. The feasibility of HCV-RT-LAMP-AuNPs-LFB was confirmed through clinical serum samples from patients with suspected HCV infections. RESULTS: An unique set of HCV-RT-LAMP primers were successfully designed targeting on the 5'UTR gene. The optimal detection process, including crude nucleic acid extraction (approximately 5 min), RT-LAMP reaction (67℃, 30 min), and visual interpretation of AuNPs-LFB results (~ 2 min), could be performed within 40 min without specific instruments. The limit of detection was determined to be 20 copies per test. The HCV-RT-LAMP-AuNPs-LFB assay exhibited high specificity and anti-interference. CONCLUSIONS: These preliminary results confirmed that the HCV-RT-LAMP-AuNPs-LFB assay is a sensitive, specific, rapid, visual, and cost-saving assay for identification of HCV. This diagnostic approach has great potential value for point-of-care (POC) diagnostic of HCV, especially in resource-challenged regions.

Biosensor-based multiple cross displacement amplification platform for visual and rapid identification of hepatitis C virus.

Hepatitis C remains a global health problem, especially in poverty-stricken areas. A rapid and sensitive point-of-care (POC) diagnostic tool is critical for the early detection and timely treatment of hepatitis C virus (HCV) infection. Here, for the first time, we reported a novel molecular diagnostic assay, termed reverse transcription multiple cross displacement amplification integrated with a gold-nanoparticle-based lateral flow biosensor (RT-MCDA-AuNPs-LFB), which was developed for rapid, sensitive, specific, and visual identification of HCV. HCV-RT-MCDA induced rapid isothermal amplification through a specific primer set targeting the 5'untranslated region gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a that are prevalent in China. The optimal reaction temperature and time for RT-MCDA-AuNPs-LFB were 68°C and 25 min, respectively. The limit of detection of the assay was 10 copies per test, and the specificity was 100% for the experimental strains. The whole detection procedure, including crude nucleic acid isolation (~5 min), RT-MCDA (68°C, 25 min), and visual AuNPs-LFB result confirmation (less than 2 min), was performed within 35 min. The preliminary results indicated that the HCV-RT-MCDA-AuNPs-LFB assay could be a valuable tool for sensitive, specific, visual, cost-saving, and rapid detection of HCV and has potential as a POC diagnostic platform for field screening and early clinical detection of HCV infection.

Efficacy of repetitive transcranial magnetic stimulation on post-stroke cognitive impairment: A systematic and a network meta-analysis.

OBJECTIVES: This study aimed to evaluate the efficacy of different repetitive transcranial magnetic stimulation (rTMS) modes in stroke patients with cognitive impairment, and to rank the best option according to the outcome measures. METHODS: Literature was searched in PubMed, Cochrane Library, Web of Science, Embase, SinoMed, China National Knowledge Infrastructure, Wanfang Database, and VIP Database, from database inception to September 2023. We included randomized controlled trials (RCTs) investigating the efficacy of all rTMS modes for post-stroke cognitive impairment. The selected studies assessed at least one of the following outcome measures: Montreal Cognitive Assessment (MoCA), Mini-Mental State Examination (MMSE), P300 latency and amplitude, and modified Barthel Index (MBI) or BI. Two researchers independently conducted data extraction. Quality assessment was performed using RevMan 5.3 software based on the Cochrane Collaboration's tool, and statistical analysis was conducted by GeMTC 0.14.3 software and Stata 17.0 software. RESULTS: The network meta-analysis included 74 RCTs with a total of 5478 patients. The best probability ranking indicated that intermittent theta burst stimulation (iTBS) was the most effective in enhancing MoCA, MMSE and MBI scores (85%, 54%, 42%, respectively), followed by 10 Hz rTMS (79%, 50%, 39%, respectively), for P300 amplitude, ≤1 Hz rTMS was ranked first (52%). CONCLUSIONS: The current limited evidence suggests that iTBS may be the optimal approach for improving cognitive and daily life abilities of stroke patients, followed by 10 Hz rTMS, ≤1 Hz rTMS may be the preferred option for enhancing P300 amplitude. TRAIL REGISTRATION: PROSPERO 2023 CRD42023424771 available from: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=424771.

[Risk Factors for Adverse Aortic Remodeling in Post-TEVAR Patients with Acute Stanford B Aortic Dissection].

OBJECTIVES: To assess morphological changes of distal aorta and the risk factors for adverse aortic remodeling inpost-TEVAR（thoracic endovascular aortic repair） patients with acute Stanford B aortic dissection. METHODS: We retrospectively investigated the patients who underwent TEVAR for a type B dissection between October 2005 and December 2015. CT angiogram (CTA) was obtained for each patients preoperatively, postoperatively and during the post-operational follow-up. Based on Criadol partition principle, we divided the aorta into descending thoracic aorta area, suprarenal abdominal aorta area, infrarenal abdominal aorta area and iliac artery area, and evaluated the distribution of aortic tears and the form of true and false lumen in different aortic partition. Univariate and multivariate logistic regression analyses were used to analyze the risk factors affecting distal aortic remodeling. RESULTS: Of 216 patients (mean follow-up (3.9±2.1) years) who were regularly followed up in our center, 47 patients (21.8%) occurred adverse remodeling in distal aorta. Univariate logistic regression indicated that abnormal aortic wall structure (Marfan's syndrome) and patent false lumen (existence of distal tears, decreased complete false lumen thrombosis) were associated with distal aortic adverse remodeling. Multivariate logistic regression showed that more tears in descending thoracic aorta area ( OR=1.36, 95% CI=1.12-1.58, P=0.005) and less tears in infrarenal abdominal aorta area ( OR=0.49, 95% CI=0.22-0.71, P<0.001) were independent risk factors affecting remodeling in distal aorta after TEVAR. CONCLUSIONS: Aortic wall structure abnormalities, a patent false lumen, more tears in descending thoracic aorta area, less tears in infrarenal abdominal aorta area are independent risk factors for adverse aortic remodeling in post-TEVAR patients with acute Stanford B aortic dissection.

Attenuation of Lipopolysaccharide-Induced Acute Lung Injury by Cyclosporine-A via Suppression of Mitochondrial DNA.

BACKGROUND Lipopolysaccharide (LPS) is generally associated with sepsis, which causes multiple system injuries and systemic inflammatory response. Mitochondrial DNA (mtDNA) is of great importance in mediation of inflammation. The aim of this study was to investigate the protective profiles of Cyclosporine-A (CsA) in LPS-induced acute lung injury (ALI) and systemic inflammation by the inhibition of mtDNA and Toll-like receptor. MATERIAL AND METHODS Twenty-four C57BL/6 mice were randomly assigned to 4 groups: a sham group (n=6); an experiment group (ALI induced through intraperitoneal injection of 10 mg/ml LPS, n=6); a low-CsA group (injection of 2.5 mg/kg of CsA 15 min after injection of LPS, n=6); and a high-CsA group (injection of 25 mg/kg of CsA 15 min after injection of LPS, n=6). Lung tissue, bronchoalveolar lavage fluid (BALF), and blood samples were collected at 6 h for further analyses. RESULTS CsA treatment significantly attenuated LPS-induced lung histopathological changes (P<.05), myeloperoxidase (MPO) activity (P<.05) and lung wet-to-dry weight ratio (P<.05). In addition, injection of CsA decreased total cells (P<.05), neutrophils (P<.05), and total protein (P<.05) in BALF and inflammatory mediators, including tumor necrosis factor-a (TNF-a, P<.05) and interleukin-6 (IL-6, P<.05) in a dose-dependent manner. A significant decrease in mtDNA was observed in the CsA group when compared with controls (P<.05). Furthermore, we demonstrated that there was a significant difference between the high-CsA group and low-CsA group in lung injury score (P<.05), mtDNA (P<.05), and MPO (P<.05). CONCLUSIONS The evidence from this study suggests that CsA attenuated lung inflammation after LPS injection, and the protective mechanism may at least in part involve decreasing the release of inflammatory cytokines and mtDNA.

Unexpected renal hemorrhage after endovascular repair of complicated type B aortic dissection: two cases report.

BACKGROUND: Thoracic endovascular aortic repair (TEVAR) is the therapeutic choice for type B aortic dissection. One of the most unfavored complications of this procedure is hemorrhage, which has a low incidence but high mortality. Renal hemorrhage (RH) after endovascular aortic repair has been rarely reported. We presented two cases of unexpected RH after TEVAR for complicated type B aortic dissection, and the potential causes, diagnosis and therapeutic management were discussed. CASE PRESENTATION: A 67-year-old female developed hypotension and progressively decrease of hemoglobin within 5 h after TEVAR for acute complicated type B dissection. Bedside ultrasonography and abdominal computed tomography angiography revealed a massive right perinephric hematoma. The right renal angiography detected multiple tortuous vascular branches with diffuse perinephric bleeding. The main trunk of right renal artery was embolized. The patient recovered uneventfully and presented with normal renal function 6 months later. Another patient was a 69-year-old male who was admitted for endovascular repair of a chronic complicated type B aortic dissection. The patient presented with hemodynamic instability early after TEVAR. Bedside ultrasonography showed a giant left retroperitoneal hematoma. The abdominal angiography revealed two active bleeding sits located in the distal branches of left renal artery. A super-selective embolization of the two arteries was performed, however the patient developed abdominal compartment syndrome and died of multiple organ failure. CONCLUSIONS: Unexpected RH after endovascular repair of aortic dissection might be associated with iatrogenic and idiopathic factors. Close surveillance and clinician's awareness of this rare complication is crucial for accurate and prompt diagnosis. Renal angiography and subsequent selective embolization of bleeding vessels are effective interventions for treating this fatal condition.

[Application of Biological Heart Valves (BHV) and Mechanical Heart Valves (MHV) in the Female Patients of Childbearing Age: a Matched Paired Study].

OBJECTIVE: To observe the efficacy and safety of biological heart valves (BHV) and mechanical heart valves (MHV) in childbearing age women (CAW) during the perinatal and short-moderate term postoperative (SMTP) periods. METHODS: There were 33 patients [(25.2±7.1) yr.] undergoing BHV replacement from September 2009 to December 2014 had completely followed-up,whose data were retrospectively collected. A 1∶4 matching study was conducted,therefore there were 132 patients undergoing MHV were included. The collected date included the clinical outcomes in the perioperative, perinatal,and SMTP period event-free survival (EFS) was determined using the Kaplan-Meier method and compared using the log-rank test. RESULTS: The average follow-up time was (5.8±3.6) years,and the two groups had similar baseline . The clinical outcomes difference of perinatal and SMTP between the two groups were not significant. There were 17 patients in BHV group and 60 in MHV group with pregnancy and birth experiences (PBE),which also showed no significant difference for adverse events both in the maternity and in the fetus. The rates of valve-related adverse events of BHV replacement patients with and without PBE were 5.9% and 0% at 3 years after the operation, and 11.8% and 5.9% after 5 years. PBE was not identified as an adverse prognostic factor for EFS (P=0.43). CONCLUSION: Either artificial BHV or MHV replacement can achieve ideal SMTP effect in CAW. BHV seems not superior to MHV. Pregnancy and birth experience will not increase the risk of BHV relevant adverse events.

Regulation of adipose-tissue-derived stromal cell orientation and motility in 2D- and 3D-cultures by direct-current electrical field.

Cell alignment and motility play a critical role in a variety of cell behaviors, including cytoskeleton reorganization, membrane-protein relocation, nuclear gene expression, and extracellular matrix remodeling. Direct current electric field (EF) in vitro can direct many types of cells to align vertically to EF vector. In this work, we investigated the effects of EF stimulation on rat adipose-tissue-derived stromal cells (ADSCs) in 2D-culture on plastic culture dishes and in 3D-culture on various scaffold materials, including collagen hydrogels, chitosan hydrogels and poly(L-lactic acid)/gelatin electrospinning fibers. Rat ADSCs were exposed to various physiological-strength EFs in a homemade EF-bioreactor. Changes of morphology and movements of cells affected by applied EFs were evaluated by time-lapse microphotography, and cell survival rates and intracellular calcium oscillations were also detected. Results showed that EF facilitated ADSC morphological changes, under 6 V/cm EF strength, and that ADSCs in 2D-culture aligned vertically to EF vector and kept a good cell survival rate. In 3D-culture, cell galvanotaxis responses were subject to the synergistic effect of applied EF and scaffold materials. Fast cell movement and intracellular calcium activities were observed in the cells of 3D-culture. We believe our research will provide some experimental references for the future study in cell galvanotaxis behaviors.

Potential Mechanism of Post-Acute Aortic Dissection Inflammatory Responses: The Role of mtDNA from Activated Platelets.

BACKGROUND: Acute aortic dissection (AD) is a lethal cardiovascular disease with severe inflammatory complications. Considering the proinflammatory properties of plasma mitochondrial DNA (mtDNA), we postulate that plasma mtDNA from activated platelets may be responsible for post-acute AD inflammatory responses. METHODS: We consecutively enrolled 68 patients with acute AD as well as matched hypertensive and healthy participants. Blood samples were collected on admission for blood routine tests, mtDNA assay, and inflammatory cytokine analysis. A computed tomography scan was used to evaluate the extent of dissections. RESULTS: Our results demonstrate that plasma mtDNA, platelet activation, and inflammatory levels were remarkably higher in acute AD patients than in hypertensive or healthy participants. These parameters were also higher in the Stanford A group than in the Stanford B group (p < 0.05). Bivariate correlation analysis demonstrated positive associations between mtDNA and inflammatory levels (tumor necrosis factor-α: r = 0.577; interleukin-6: r = 0.632), mtDNA and platelet activation (r = 0.642), and platelet activation and the extent of dissection (r = 0.635). CONCLUSION: Our study suggests that acute AD-induced tunica media exposure causes platelet activation, which leads to the initiation of inflammatory responses via the release of mtDNA into the circulation. Our study provides a novel fundamental basis and a potential therapeutic target for the prevention and treatment of post-AD inflammatory responses.

[The Association Between Bone Morphogenic Protein-4 and in-situ Saphenous Vein Graft Pathology in Diabetic Patients Undergoing Coronary Revascularization].

OBJECTIVES: This study was designed to assess the expression levels of bone morphogenic protein-4 (BMP4) in saphenous veins harvested from diabetic patients undergoing coronary artery bypass grafting (CABG), and to investigate its association with in-situ morphological and pathological changes within the venous wall. METHODS: From January 2013 to December 2014, forty patients with type 2 diabetes mellitus (DM) and risk factors matched non-DM controls (n =40) were enrolled prospectively. Of the 40 DM patients, 24 had noninsulin-dependent diabetes (NIDM) and the remaining 16 had insulin-dependent diabetes (IDM). Segments of saphenous vein without surgical dilatation from these 80 patients were obtained. Vessel wall characteristics were evaluated by hematoxylin-eosin (HE) staining, and the expression and distribution of BMP4 was assessed by Western blot assay and immunohistochemistry analysis. RESULTS: The intimal and medial thickness of the saphenous veins harvested from DM patients were higher than those from non-DM controls. Compared with non-DM patients, the expression level of BMP4 was significantly elevated in diabetic veins ( P<0.05), and BMP4 was highly expressed in smooth muscle cells located in the medial layer. Moreover, the expression levels of BMP4 in diabetic veins were significantly correlated with intimal thickness (r =0.655, P<0.01), intimal area (r =0.684, P<0.01), medial thickness (r =0.642, P<0.01) and medial area (r =0.692, P<0.01). CONCLUSIONS: The pre-existing intimal and medial hyperplasia were significantly correlated with the high expression levels of BMP4 in saphenous veins harvested from diabetic patients, implying a potential role of BMP4 in the progression of vein graft stenotic diseases in this cohort of post-CABG patients. Future studies were warranted in investigating novel therapeutic strategy targeting at BMP4 for improving long-term vein graft patency.

[Proximal Aorta Reconstruction for Type A Aortic Dissection Patients without Intimal Tear in Aortic Arch: the Early and Mid-outcomes].

OBJECTIVE: To evaluate the early and mid-outcomes of proximal aorta reconstruction for type A aortic dissection (AAD) patients without intimal tear in aortic arch, and assess the safety and efficacy of this surgical strategy. METHODS: From January 2010 to February 2013, there were 23 AAD patients without intimal tear in the aortic arch received proximal aorta reconstruction surgery. Clinical data of these patients were analyzed retrospectively, the mean age was (48.04 ± 12.37) years old (21-73 yr.). Twelve cases were acute aortic dissection, the others were chronic dissection. Bentall surgery was performed for 13 cases, Cabrol surgery for 2 cases, Wheat surgery for 1 case, ascending aorta replacement and aortic valve repair was employed for 1 patient, simple ascending aorta replacement for 6 cases. The patients received follow-up every 3 to 6 months after the surgery. RESULTS: The duration of CPB time was (182.83 ± 36.98) min, cardiac arrest time was (111.87 ± 18.82) min, circulatory arrest time was (24.22 ± 6.38) min. The complications were lung infection (4 cases, 17.4%), tracheotomy (2 cases), peritoneal dialysis (1 case), 2 cases suffered transient neurological dysfunction. None stroke, paralysis, and permanent neurological dysfunction occurred. All the patients were discharged. Mean time of follow-up was (38.35 ± 11.95) months (18-56 months). All patients were alive and return to normal life, the proportion of false lumen closure was 65.22% (15 cases). None patients need secondary surgery. CONCLUSION: Proximal aorta reconstruction is safe and effective for AAD patient without intimal tear in aortic arch, the operation strategy can be used individually.

MiR-489 modulates cisplatin resistance in human ovarian cancer cells by targeting Akt3.

MicroRNAs are a conserved class of small noncoding RNA molecules that harbour the capacity to regulate protein-coding gene expression at the post-transcriptional level. In the current study, we show that miR-489 is downregulated in cisplatin (CDDP)-resistant ovarian cancer cells, SKOV3/CDDP and OVCAR3/CDDP cells. MiR-489 overexpression results in an inhibition of SKOV3 and OVCAR3 cell survival and cell growth after CDDP treatment and an induction of cell apoptosis. Inhibition of miR-489 yields the opposite results. In addition, miR-489 overexpression increases the sensitivity of SKOV3/CDDP and OVCAR3/CDDP cells to CDDP and inhibits their colony number. Akt3 is validated as a direct target of miR-489 in SKOV3, OVCAR3, SKOV3/CDDP and OVCAR3/CDDP cells. In addition, miR-489 suppresses Akt3 protein expression by binding sites on its 3'UTR. Knockdown of Akt3 results in a similar effect as that because of miR-489 overexpression; importantly, Akt3 silencing rescues the functions induced by miR-489. Furthermore, we also use the Akt3 inhibitor, MK-2206 2HCl, to determine the role of Akt3 in CDDP resistance. Our study showed that MK-2206 2HCl increased the sensitivity of SKOV3/CDDP and OVCAR3/CDDP cells to CDDP. Taken together, our results indicate that miR-489 inhibited CDDP resistance and cell growth, and promotes apoptosis by suppressing Akt3 expression. Furthermore, the identification of a novel miR-489-based pathway in CDDP-resistant ovarian cancer will facilitate the development of therapeutic strategies.

The relationship between gastrointestinal symptoms in FGID patients and D-type personality and emotion regulation strategies.

This study examines the relationship between gastrointestinal symptoms in patients with functional gastrointestinal disorders (FGIDs) and type D personality traits, as well as emotion regulation strategies. Analyzing a diverse group of FGID patients, we uncover significant effects of gender and age on gastrointestinal symptoms. Negative Affectivity emerges as a key predictor, positively associated with symptom severity, whereas Social Inhibition correlates negatively with Abdominal Pain. Additionally, our findings suggest that the expressive suppression strategy predicts heightened gastrointestinal symptoms, whereas cognitive reappraisal predicts lower levels of certain symptoms. These findings provide valuable insights for precise diagnosis and tailored treatments of FGIDs. Further research is warranted to explore underlying mechanisms and inform evidence-based interventions.

Unique Approach for Isolating Rat Bone Marrow Neutrophils with Comparable Capacity.

The primary aim of this research was to develop a reliable and efficient approach for isolating neutrophil extracellular traps (NETs) from rat bone marrow. This effort arose due to limitations associated with the traditional method of extracting NETs from peripheral blood, mainly due to the scarcity of available neutrophils for isolation. The study revealed two distinct methodologies for obtaining rat neutrophils from bone marrow: a streamlined one-step procedure that yielded satisfactory purification levels, and a more time-intensive two-step process that exhibited enhanced purification efficiency. Importantly, both techniques yielded a substantial quantity of viable neutrophils, ranging between 50 to 100 million per rat. This efficiency mirrored the results obtained from isolating neutrophils from both human and murine sources. Significantly, neutrophils derived from rat bone marrow exhibited comparable abilities to secrete NETs when compared with neutrophils obtained from peripheral blood. However, the bone marrow-based method consistently produced notably larger quantities of both neutrophils and NETs. This approach demonstrated the potential to obtain significantly greater amounts of these cellular components for further downstream applications. Notably, these isolated NETs and neutrophils hold promise for a range of applications, spanning the realms of inflammation, infection, and autoimmune diseases.