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Long-term explant culture: an improved method for consistently harvesting homogeneous populations of keloid fibroblasts
Li, J; Zou, YQ; Wang, S; Guo, SK; Huang, ZS; Huo, R
BIOENGINEERED 2022年 13卷1期 页码:1565- 影响因子:1.87
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Explant culture is a more suitable method than enzyme digestion for the isolation of keloid fibroblasts (KFs), but it has a longer isolation period. In this study, we propose a long-term explant culture method. Unlike in the conventional explant culture method, we continued culturing explants to isolate KFs rather than discarding them during passage. We demonstrated that keloid explants could be cultured for more than 4 months to continuously yield enriched KFs, and the KFs from the repeatedly cultured explants had shorter isolation times. The biological behavior and fibrotic phenotypic characteristics of the KFs from the explants cultured long term were investigated, and no statistical differences were found compared with the KFs from the original explants. In conclusion, the long-term explant culture method was shown to be efficient for harvesting a large, homogeneous population of KFs. The high efficiency as well as ease of operation and sample saving make this method convenient for researchers working with KFs.
Hypoxic preconditioning reduces NLRP3 inflammasome expression and protects against cerebral ischemia/reperfusion injury
Pang, YQ; Yang, J; Jia, CM; Zhang, R; Pang, Q
NEURAL REGENERATION RESEARCH 2022年 17卷2期 页码:395-
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Hypoxic preconditioning can protect against cerebral ischemia/reperfusion injury. However, the underlying mechanisms that mediate this effect are not completely clear. In this study, mice were pretreated with continuous, intermittent hypoxic preconditioning; 1 hour later, cerebral ischemia/reperfusion models were generated by middle cerebral artery occlusion and reperfusion. Compared with control mice, mice with cerebral ischemia/reperfusion injury showed increased Bederson neurological function scores, significantly increased cerebral infarction volume, obvious pathological damage to the hippocampus, significantly increased apoptosis; upregulated interleukin-1 beta, interleukin-6, and interleukin-8 levels in brain tissue; and increased expression levels of NOD-like receptor family pyrin domain containing 3 (NLRP3), NLRP inflammasome-related protein caspase-1, and gasdermin D. However, hypoxic preconditioning significantly inhibited the above phenomena. Taken together, these data suggest that hypoxic preconditioning mitigates cerebral ischemia/reperfusion injury in mice by reducing NLRP3 inflammasome expression. This study was approved by the Medical Ethics Committee of the Fourth Hospital of Baotou, China (approval No. DWLL2019001) in November 2019.
Implementation of a temperature bundle improves admission hypothermia in very-low-birth-weight infants in China: a multicentre study
Wang, L; Liu, ZJ; Liu, FM; Yu, YH; Bi, SY; Li, B; Xu, HY; Yang, CY
BMJ OPEN QUALITY 2022年 11卷2期 页码:-null
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Background Hypothermia is a common problem that is associated with increased mortality and morbidity among preterm infants, especially in China. The objective of this study was to evaluate the efficacy of a targeted quality improvement (QI) project that applied hypothermia prevention measures for very-low-birth-weight (VLBW) infants in three tertiary neonatal intensive care units (NICUs) in China. Problem Between January 2018 and December 2018, we conducted a prospective analysis and found that the incidence of AH was 88.2% among VLBW infants. Methods The study enrolled preterm infants born at less than 32 weeks' gestation with a VLBW of less than 1500 g who were delivered at three academic tertiary-care hospitals between January 2018 and December 2019. The primary outcome measure was the incidence of hypothermia. The outcomes of the pre-QI group (1 January-31 December 2018) were compared with those of the post-QI group (1 January-31 December 2019). Interventions Based on the literature, our preliminary findings and the needs of each unit, a temperature bundle that included a transport incubator, prewarmed hats, polyethylene wrap, team training and education, and temperature documentation and workflows were implemented in consecutive plan-do-study-act cycles. Results Of the 530 VLBW infants, 235 infants (36.9%) belonged to the pre-QI group, and 295 infants (46.4%) belonged to the post-QI group. The incidence of hypothermia decreased significantly, from 92.3% to 62% (p<0.001), and the mean body temperature on admission to the NICU increased significantly, from 35.5 degrees C to 36 degrees C +/- 0.7 degrees C (p<0.001). There was one case of hyperthermia during the study period. Infants in the post-QI group had a lower mortality rate (16.1% vs 8.8%, p=0.01). Conclusions Targeted interventions can dramatically reduce admission hypothermia and improve the outcome of VLBW infants in China.
Use of ultrasonography to evaluate early outcomes of reduction in developmental dysplasia of the hip
Luan, QH; Teng, JB; Shi, M; Li, TY; Sun, B; Wang, YZ; Lin, XT; Ban, YG
PEDIATRIC RADIOLOGY 2022年 页码:-null
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Objective To compare pubo-femoral distance (PFD) in normal hips and those treated for developmental dysplasia of the hip (DDH) and to investigate the value of ultrasonography from the medial hip in early follow-up of dislocated DDH after reduction. Materials and methods This study included 58 infants (49 females) with DDH who presented with 65 dislocated hips (51 unilateral and 7 bilateral). Dislocation was treated by closed reduction for 53 and open reduction for 12 hips. Ultrasonography on the medial side of the hip was performed within 1-2 weeks and 4 weeks after reduction. The distance from the pubic bone to the femoral head (PFD) was measured to assess the reduction and stability of the femoral head and compared to that on the contralateral side (control) in cases of unilateral DDH. Results The PFD value for the normal group (2.9 +/- 0.4 mm) was significantly less than that for the closed reduction group (4.9 +/- 2.8 mm, P<0.001) and that for the open reduction group (4.4 +/- 1.6 mm; P=0.02), but no difference in the PFD was observed between the closed reduction and the open reduction groups (P=0.73). Despite successful reduction, the PFD values in the successful reduction group remained higher than those of the normal hips. Conclusion PFD measurement by ultrasonography of the medial hip can be used to evaluate the effectiveness of reduction procedures in DDH. The clinical implications of post-reduction ultrasound evaluation in the diagnosis and long-term follow-up of outcomes require further research.
Eculizumab treatment for myasthenia gravis subgroups: 2021 update
Jiao, L; Li, HH; Guo, SG
JOURNAL OF NEUROIMMUNOLOGY 2022年 362卷 页码:-null
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Eculizumab is a recombinant humanized monoclonal antibody that targets the complement protein C5, inhibiting its cleavage into C5a and C5b and ultimately preventing the formation of C5b-9 membrane attack complex (MACs), thereby protecting the neuromuscular junction from the damage of complement activation. In 2017, eculizumab became the second FDA-approved medication for AchR-positive generalized myasthenia gravis (gMG) patients based on the successful results of a randomized, double-blinded, placebo-controlled, phase 2, phase 3 study (the REGAIN trial) and its open-label extension study. Despite the efficacy of eculizumab in treating AchR antibody-positive refractory gMG was demonstrated in the REGAIN study, there is few information on its efficacy in other subgroup of MG patients including seronegative MG, thymoma-associated MG and MG crisis. This narrative review summarizes current clinical studies of eculizumab in these refractory gMG patients, with a focus on the therapeutic efficacy and tolerability in different subgroup of MG.
Knockdown of lncRNA NEAT1 suppresses proliferation and migration, and induces apoptosis of cervical cancer cells by regulating the miR-377/FGFR1 axis
Geng, F; Jia, WC; Li, T; Li, N; Wei, W
MOLECULAR MEDICINE REPORTS 2022年 25卷1期 页码:-null
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To investigate the role of NEAT1 and the microRNA (miR)-377/fibroblast growth factor receptor 1 (FGFR1) axis in cervical cancer (CC), the expression levels of NEAT1, FGFR1 and miR-377 were detected in CC tissues and cell lines. NEAT1 or FGFR1 was knocked down by transfection with short hairpin RNA (sh)-NEAT1 or sh-FGFR1, and miR-377 was overexpressed by transfection with miR-377 mimics in HeLa and C33A cells. Cell viability and migration were measured using MTT and Transwell assays, respectively. Cell apoptosis was determined by flow cytometry. A dual luciferase reporter assay was performed to confirm the presence of binding sites between miR-377 and FGFR1. The results revealed that the expression levels of NEAT1 and FGFR1 were significantly elevated, whereas miR-377 expression was markedly decreased in CC tissues and cell lines. In HeLa and C33A cells, after NEAT1 knockdown, miR-377 expression was increased, cell viability and migration were inhibited, and apoptosis was induced. Similarly, silencing FGFR1 inhibited cell viability and migration, and induced apoptosis of HeLa and C33A cells. A dual luciferase reporter gene assay verified a targeting relationship between NEAT1 and miR-377. Inhibition of miR-377 or overexpression of FGFR1 reversed the effects of NEAT1 knockdown on cell function in HeLa and C33A cells. Moreover, a dual luciferase reporter assay confirmed that FGFR1 was a direct target of miR-377. In conclusion, suppression of NEAT1 inhibited cell viability and migration, and promoted apoptosis of CC cells, and these effects were achieved through regulation of the miR-377/FGFR1 axis.
Head and Neck Squamous Cell Carcinoma: Risk Factors, Molecular Alterations, Immunology and Peptide Vaccines
Sun, Z; Sun, XD; Chen, ZW; Du, J; Wu, YH
INTERNATIONAL JOURNAL OF PEPTIDE RESEARCH AND THERAPEUTICS 2022年 28卷1期 页码:-null
无关键词信息
Head and neck squamous cell carcinoma (HNSCC) arises from the epithelial lining of the oral cavity, hypopharynx, oropharynx, and larynx. There are several potential risk factors that cause the generation of HNSCC, including cigarette smoking, alcohol consumption, betel quid chewing, inadequate nutrition, poor oral hygiene, HPV and Epstein-Barr virus, and Candida albicans infections. HNSCC has causative links to both environmental factors and genetic mutations, with the latter playing a more critical role in cancer progression. These molecular changes to epithelial cells include the inactivation of cancer suppressor genes and proto-oncogenes overexpression, resulting in tumour cell proliferation and distant metastasis. HNSCC patients have impaired dendritic cell (DC) and natural killer (NK) cell functions, increased production of higher immune-suppressive molecules, loss of regulatory T cells and co-stimulatory molecules and major histocompatibility complex (MHC) class Iota molecules, lower number of lymphocyte subsets, and a poor response to antigen-presenting cells. At present, the standard treatment modalities for HNSCC patients include surgery, chemotherapy and radiotherapy, and combinatorial therapy. Despite advances in the development of novel treatment modalities over the last few decades, survival rates of HNSCC patients have not increased. To establish effective immunotherapies, a greater understanding of interactions between the immune system and HNSCC is required, and there is a particular need to develop novel therapeutic options. A therapeutic cancer vaccine has been proposed as a promising method to improve outcome by inducing a powerful adaptive immune response that leads to cancer cell elimination. Compared with other vaccines, peptide cancer vaccines are more robust and specific. In the past few years, there have been remarkable achievements in peptide-based vaccines for HNSCC patients. Here, we summarize the latest molecular alterations in HNSCC, explore the immune response to HNSCC, and discuss the latest developments in peptide-based cancer vaccine strategies. This review highlights areas for valuable future research focusing on peptide-based cancer vaccines.
Clinical significance and effect of lncRNA BBOX1-AS1 on the proliferation and migration of lung squamous cell carcinoma
Zhang, Y; Wang, X; Cheng, XK; Zong, YY; He, RQ; Chen, G; Qin, YJ
ONCOLOGY LETTERS 2022年 23卷1期 页码:-null
无关键词信息
Long non-coding RNAs (lncRNAs) have a role in the occurrence and development of lung squamous cell carcinoma (LUSC). lncRNA gamma-butyrobetaine hydroxylase 1 (BBOX1)-antisense 1 (AS1) may contribute to disease development. However, there are no studies on the role of BBOX1-AS1 in LUSC to date. In the present study, an in-house gene microarray analysis was performed to detect the differentially expressed lncRNAs and mRNAs between three pairs of LUSC and normal lung tissues. Only one lncRNA, BBOX1-AS1, was differentially expressed in the in-house microarray and The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO) and ArrayExpress databases. Reverse transcription-quantitative PCR (RT-qPCR) was then performed and the original RNA-sequencing data from the TCGA, GEO and ArrayExpress datasets were used to determine the expression and clinical value of BBOX1-AS1 in LUSC. In addition, a Cell Counting Kit-8 assay, cell cycle analysis and scratch assay were performed to explore whether BBOX1-AS1 expression affected the proliferation and migration of LUSC cells in vitro. The results of the RT-qPCR analysis and data obtained from the TCGA database, GEO datasets, in-house gene microarray and standard mean deviation analysis all supported the upregulated expression level of BBOX1-AS1 in LUSC. Furthermore, silencing of BBOX1-AS1 inhibited the proliferation and migration of LUSC cells according to in vitro assays. In addition, the cells were arrested in S-phase after knockdown of BBOX1-AS1. In conclusion, the expression level of BBOX1-AS1 was upregulated in LUSC tissues. BBOX1-AS1 may exert an oncogenic effect on LUSC by regulating various biological functions. However, additional functional experiments should be performed to verify the exact mechanism.
Electrochemical detection of ACE2 as a biomarker for diagnosis of COVID-19 and potential male infertility
Liu, S; Han, LP; Li, JL; Li, H
BIOSENSORS & BIOELECTRONICS 2022年 198卷 页码:-null
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Knowing how heavily the body is burdened by SARS-CoV-2 infection is all important to avoid tragic outcomes. This demands fast and convenient assays with minimum requirement for instruments and reagents. Therefore, a short synthetic peptide is developed to perform direct serum assay, using portable hand-held potentiostat, in a reagent-less manner. The target is angiotensin converting enzyme 2 (ACE2), a protein secreted by the body into the blood to restrict viral invasion. Specifically, under electrochemical potential scanning, the peptide can covalently capture ACE2 from the serum, and then form a covalent gel-like 2D protein network with the serum proteins, in an ACE2-specific fashion. This formation of a covalent biosensing complex enables sensitive detection in serum samples of SARS-CoV-2 infected patients. The detected serum level of ACE2 can not only serve as an index of viral load, but may also hint at the associated risk of potential male infertility. These results may point to field application of this simple design in the clinical practice in treating COVID-19 in the near future.
Randomized Controlled Trial Comparing the Short-term Outcomes of Enhanced Recovery After Surgery and Conventional Care in Laparoscopic Distal Gastrectomy (GISSG1901)
Tian, YL; Cao, SG; Liu, XD; Li, LP; He, QS; Jiang, LX; Wang, XJ; Chu, XQ; Wang, H; Xia, LJ; Ding, YL; Mao, WZ; Hui, XZ; Shi, YR; Zhang, HH; Niu, ZJ; Li, ZQ; Jiang, HT; Kehlet, H; Zhou, YB
ANNALS OF SURGERY 2022年 275卷1期 页码:E15-
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Objective: This study aimed to compare the effects of ERAS and conventional programs on short-term outcomes after LDG. Summary of Background Data: Currently, the ERAS program is broadly applied in surgical areas. Although several benefits of LDG with the ERAS program have been covered, high-level evidence is still limited, specifically in advanced gastric cancer. Methods: The present study was designed as a randomized, multicenter, unblinded trial. The enrollment criteria included histologically confirmed cT2-4aN0-3M0 gastric adenocarcinoma. Postoperative complications, mortality, readmission, medical costs, recovery, and laboratory outcomes were compared between the ERAS and conventional groups. Results: Between April 2019 and May 2020, 400 consecutive patients who met the enrollment criteria were enrolled. They were randomly allocated to either the ERAS group (n = 200) or the conventional group (n = 200). After excluding patients who did not undergo surgery or gastrectomy, 370 patients were analyzed. The patient demographic characteristics were not different between the 2 groups. The conventional group had a significantly longer allowed day of discharge and postoperative hospital stay (6.96 vs 5.83 days, P < 0.001; 8.85 vs 7.27 days, P < 0.001); a longer time to first flatus, liquid intake and ambulation (3.37 vs 2.52 days, P < 0.001; 3.09 vs 1.13 days, P < 0.001; 2.85 vs 1.38 days, P < 0.001, respectively); and higher medical costs (6826 vs 6328 $, P = 0.027) than the ERAS group. Additionally, patients in the ERAS group were more likely to initiate adjuvant chemotherapy earlier (29 vs 32 days, P = 0.035). There was no significant difference in postoperative complications or in the mortality or readmission rates. Regarding laboratory outcomes, the procalcitonin and C-reactive protein levels on postoperative day 3 were significantly lower and the hemoglobin levels on postoperative day 5 were significantly higher in the ERAS group than in the conventional group. Conclusion: The ERAS program provides a faster recovery, a shorter postoperative hospitalization length, and lower medical costs after LDG without increasing complication and readmission rates. Moreover, enhanced recovery in the ERAS group enables early initiation of adjuvant chemotherapy.
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