AIM: To construct a prokaryotic expression system of pET15b-PEP-1-SOD1, and to determine whether the expressed PEP-1-SOD1 fusion protein can be transduced into cells. METHODS: SOD1 cDNA was amplified by PCR using pBluescript II SK-SOD1 as template, and digested with restriction endonuclease to construct recombinant plasmids pET15b-SOD1 and pET15b-PEP-1-SOD1, followed by being respectively transformed into E.coli to produce SOD1 and PEP-1-SOD1 fusion proteins, which were purified and added in cultured endothelial cells of human umbilical vein in vitro. RESULTS: The highly purified and active SOD1 and PEP-1-SOD1 fusion proteins were obtained.The PEP-1-SOD1 fusion protein could enter endothelial cells of human umbilical vein in a time- and dose-dependent manner when added exogenously in a culture media. Once inside the cells, PEP-1-SOD1 protein was enzymatically stable for 48 h. CONCLUSION: PEP-1-SOD1 fusion protein can be transduced effectively into endothelial cells of human umbilical vein, which provides a basis for future study involving direct delivery of SOD1 protein into tissues for therapy.
Purpose: To explore the preliminary safety and efficacy of the Shurui single-port (SP) surgical robot in partial nephrectomy (PN). Methods: This study prospectively enrolled patients with T1a renal tumors who met the inclusion criteria from February to July 2022 in The First Affiliated Hospital School of Medicine Zhejiang University. The operative outcomes and perioperative data, including clinical and histological data, were prospectively collected and analyzed. Results: A total of 13 patients were included in this study, including 7 males and 6 females. The median age was 53 (33–74) years, and the average body mass index was 24.9 ± 4.2 kg/m2. There were 6 cases of left kidney tumors and 7 cases of right kidney tumors in the 13 patients. The average tumor diameter was 1.9 ± 0.9 cm. In all operations, the diseased tissue was removed according to the established surgical plan. The average warm ischemia time was 26.2 ± 9.7 minutes; the average device docking time was 3.6 ± 1.8 minutes; and the average robotic arm operation time was 124.7 ± 40.4 minutes. All operations were successfully completed; there was no conversion to open surgery during the operation; and no operation holes were added. The National Aeronautics and Space Administration Task Load Index (NASA-TLX) score was 26.3 ± 2.6 points, and no device-related adverse events occurred during the operation. The median time to discharge was 6 days (range, 4–11 days). Postoperative pathological examination showed that all tumor margins were negative. There were no Clavien grade ≥3 surgical complications in any of the patients during the perioperative period or at the 1-month postoperative follow-up. Conclusion: The new SP surgical robot system is safe, effective, flexible, and stable for application in PN.
Serum cotinine is a common biomarker for smoking and secondhand smoke (SHS) exposure, but it can be affected by the activity of nicotine-metabolizing enzymes. This study investigated the influence of CYP2A6*4 genotypes on serum cotinine among nonsmoking pregnant women.We analyzed the data from 545 Chinese nonsmoking pregnant women in a case-control study on SHS exposure and birth outcomes in southern China. Participants self-reported their status and duration of SHS exposure during pregnancy right after delivery in hospital. Research staff used polymerase chain reaction to genotype CYP2A6*4 and enzyme-linked immunosorbent assay to measure cotinine levels in maternal serum samples collected before delivery. We stratified women by their self-reported SHS exposure status and CYP2A6*4 genotypes and then compared their median levels of serum cotinine.Among women who self-reported non-SHS exposure (n = 317), the median serum cotinine levels were 2.83ng/ml for those with CYP2A6*1/*1 genotype, 1.39ng/ml for CYP2A6*1/*4, and 0.77ng/ml for CYP2A6*4/*4, respectively. Among women who self-reported SHS exposure (n = 228), the median cotinine levels were 3.32ng/ml for those with CYP2A6*1/*1 genotype, 2.38ng/ml for CYP2A6*1/*4, and 1.56ng/ml for CYP2A6*4/*4, respectively. Strikingly, self-reported SHS-exposed women with CYP2A6*1/*4 or CYP2A6*4/*4 genotype had significantly lower (rather than higher) median cotinine levels than self-reported non-SHS-exposed women with CYP2A6*1/*1 genotype (p = .012).CYP2A6*4 genotype is associated with lower serum cotinine among Chinese nonsmoking pregnant women. Measuring CYP2A6*4 genotype may help to improve the validity of SHS exposure measurement by serum cotinine in pregnant women and possibly also in other nonpregnant populations.
Objective: Low-frequency oscillations (LFOs) observed in near-infrared spectroscopy (NIRS) are closely related to autonomic physiological processes and may serve as useful indicators for detecting and monitoring circulatory dysfunction. The aim of this study was to investigate the difference of symmetrical LFOs in healthy subjects, ischemic stroke patients and peripheral atherosclerotic patients.Materials and Methods: In this study, healthy controls, ischemic stroke patients and peripheral atherosclerotic patients completed a resting-state LFOs detection experiment. LFOs were collected simultaneously at peripheral right and left earlobes, fingertips and toes, and performed coherence and phase shift analyses processing.Results: The results showed that LFOs were better able to differentiate between healthy individuals and those with peripheral artery disease compared with stroke patients. The significant differences between healthy and stroke patients may be related to peripheral vascular disease in stroke patients. Asymmetrical LFOs in peripheral atherosclerosis patients may be used to indicate location of vascular lesions, to an extent.Conclusions: The reduced coherence coefficients and increased phase shifts of LFOs in peripheral atherosclerosis patients support that symmetrical LFOs can not only identify vascular injury disease, but also provide baseline references for exploring the overall feedback of pericentral interactions. LFOs have the potential to provide valuable information about patients that assist researchers and clinicians in identifying specific circulatory damage subgroups.
Background Genetic variation plays an extremely important pathogenic role in the development of venous thromboembolism (VTE). Genetic protein S (PS) deficiency caused by PROS1 gene mutation is an important risk factor for hereditary thrombophilia. Case introduction In this case, we report a 28-year-old male patient who developed a severe pulmonary embolism during his visit. The patient had experienced one month of chest pains, coughing and hemoptysis symptoms. CTPA confirmed an acute pulmonary embolism with multiple filling defects in both pulmonary arteries. Ultrasound showed no thrombosis in the veins of both lower limbs. The patient's father and grandfather have a history of lower limb venous thrombosis. The patient was diagnosed with acute pulmonary embolism and pneumonia. The serum PS level significantly decreased (detection result: 10%, normal range: 77–143). Gene sequencing revealed a heterozygous missense mutation in PROS1 c.76+2_76+3del (base deletion), and further testing revealed that the genetic variation originated from his father. The patient was treated with heparin anticoagulant therapy, catheter thrombus aspiration, and catheter thrombolysis. After treatment, the patient's chest pain symptoms were relieved, and there were no symptoms such as difficulty breathing. On the 7th day of admission, the patient was transferred to a general hospital for further treatment. Conclusion Hereditary thrombophilia caused by mutations in the PROS1 (c.76+2_76+3del) gene is extremely rare. In clinical practice, heparin and rivaroxaban treatment are beneficial.
To explore the mediation effects of maternal serum interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) on the association between environmental tobacco smoke (ETS) exposure during pregnancy and low birth weight (LBW) at term.ETS exposure, birth weight, blood sample and covariates were collected from 195 pregnant women delivered term LBW infants and 195 controls delivered normal birth weight infants in two Maternity and Child Hospitals in Guangdong, China. Maternal serum IL-1β and TNF-α were measured by flow cytometry. Logistic regression models and path analyses explored the mediation effects of maternal IL-1β and TNF-α on the association between ETS exposure and LBW.LBW was significantly associated with maternal ETS exposure (OR = 2.14 (95% CI =1.06-4.32)). TNF-α and IL-1β were significantly associated with both LBW (OR = 1.87 (1.41-2.47) and OR = 1.53 (1.14-2.05)) and ETS (β = 0.32 (0.04-0.60) and β = 0.27 (0.05-0.49)). Traditional mediation analyses indicated the separate mediation effect of TNF-α and IL-1β was 32.2% and 24.6%, respectively. Path analysis revealed the combined mediation effects of TNF-α and IL-1β as 29.4% in the pathway from ETS exposure to LBW.Maternal serum IL-1β and TNF-α may play a mediating role in the association between maternal ETS exposure during pregnancy and term LBW.
Acute respiratory distress syndrome (ARDS) is a common life-threatening critical illness with high mortality. Fusu mixture (FSM) can improve the mechanical ventilation in ARDS patients. However, the detailed pharmacological mechanisms and active substances of FSM are still unclear. This study aimed to explore the potential pharmacological mechanisms of FSM for treating ARDS and its chemical compositions.A lipopolysaccharide (LPS)-induced ARDS mouse model was established, and the mice subsequently received FSM (50 mg/kg) orally for 5 days. Then, the blood samples and lung tissues were collected. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in serum, and histopathology examinations were applied to analyze the inflammatory response of lung tissues in ARDS mice. In addition, protein expressions of aquaporin 5 (AQP-5), surfactant-associated protein C (SP-C), and Notch1 were detected by western blot assays and immunohistochemical (IHC) examination. In addition, the chemical compositions of FSM were analyzed by high performance liquid chromatography (HPLC), using standard reference agents.After LPS induction, the serum levels of IL-6 and TNF-α in ARDS mice were significantly increased (P<0.01, vs. Control), and FSM significantly reduced these 2 pro-inflammatory cytokines (IL-6 and TNF-α) compared to the model mice (P<0.01). Histopathology examinations showed FSM significantly attenuated the inflammatory responses in lung tissues. Furthermore, after FSM treatment, the SP-C and AQP-5 were significantly increased, compared to the Model mice (P<0.01), and FSM also up-regulated the Notch1 expressions in lung tissues of ARDS mice (P<0.001, vs. Model).Collectively, it is suggested that FSM alleviates inflammatory reactions and promotes the proliferation of alveolar epithelial cells in LPS-induced ARDS mice via regulation of SP-C, AQP-5, and Notch1 in lung tissues.
Objective: We sought to effectively alleviate the emotion of individuals with anxiety and depression, and explore the effects of aerobic exercise on their emotion regulation. Functional near-infrared spectroscopy (fNIRS) brain imaging technology is used to monitor and evaluate the process of aerobic exercise and imagination that regulates emotion. Approach:Thirty participants were scored by the state-trait anxiety inventory (STAI) and profile of mood states (POMS), and fNIRS images were collected before, after, and during aerobic exercise and motor imagery. Then, the oxygenated hemoglobin (HbO), deoxygenated hemoglobin (HbR), and total hemoglobin (HbT) concentrations and their average value were calculated, and the ratio of HbO concentration in the left and right frontal lobes was determined. Spearman's correlation coefficient was used to calculate the correlation between variations in the average scores of the two scales and in blood oxygen concentrations. Results: In comparison with motor imagery, STAI, and POMS scores decreased after 20 min of aerobic exercise. The prefrontal cortex had asymmetry and laterality (with the left side being dominant in emotion regulation). The increase in hemoglobin concentration recorded by fNIRS was negatively correlated with STAI and POMS scores. Aerobic exercise has a good effect on emotion regulation. Significance:The study showed that portable fNIRS could be effectively used for monitoring and evaluating emotion regulation by aerobic exercise. This study is expected to provide ideas for constructing fNIRS-based online real-time monitoring and evaluation of emotion regulation by aerobic exercise.
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