Objective To investigate the role and mechanism of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) in the activation of aortic valve interstitial cells (AVICs) in aortic stenosis. Methods Isolating primary AVICs and stimulating their activation with transforming growth factor β1 (TGF-β1, 30 ng/mL), the expression of PGC-1α was detected. The activation of AVICs induced by TGF-β1 was observed after overexpression of PGC-1α by adenovirus or inhibition of PGC-1α function by GW9662. The possible downstream molecular mechanism of PGC-1α in AVICs activation was screened. Finally, the phenotype was further verified in primary human AVICs. Results The expression of PGC-1α decreased after the activation of AVICs induced by TGF-β1 (control group: 1.00±0.18; 24 h: 0.31±0.10; 48 h: 0.32±0.06; 72 h: 0.20±0.07; P<0.05). Specific overexpression of PGC-1α by adenovirus inhibited the activation of AVICs induced by TGF-β1 stimulation (periostin: 3.17±0.64 vs. 1.45±0.54, P<0.05; α-smooth muscle actin: 0.77±0.11 vs. 0.28±0.06, P<0.05). On the contrary, inhibition of PGC-1α function by GW9662 promoted the activation of AVICs (periostin: 2.20±0.68 vs. 7.99±2.50, P<0.05). Subsequently, it was found that PGC-1α might inhibit the activation of AVICs through downregulating the expression of calcium/calmodulin-dependent protein kinase (CAMK1δ) (0.97±0.04 vs. 0.74±0.11, P<0.05), and downregulating the expression of CAMK1δ alleviated the activation of AVICs (periostin: 1.76±0.11 vs. 0.99±0.20, P<0.05). The possible mechanism was that the activation of mammalian target of rapamycin (mTOR) signaling pathway was inhibited by reducing the accumulation of reactive oxygen species (ROS) (778.3±139.4 vs. 159.3±43.2, P<0.05). Finally, the protective effect of PGC-1α overexpression was verified in the activated phenotype of human AVICs (periostin: 2.73±0.53 vs. 1.63±0.14, P<0.05; connective tissue growth factor: 1.27±0.04 vs. 0.48±0.09, P<0.05). Conclusions The expression of PGC-1α significantly decreases during the activation of AVICs induced by TGF-β1. The overexpression of PGC-1α significantly inhibites the activation of AVICs, suggesting that PGC-1α plays a protective role in the activation of AVICs. The possible mechanism is that PGC-1α can inhibit the activation of CAMK1δ-ROS-mTOR pathway. In conclusion, interventions based on PGC-1α expression levels are new potential therapeutic targets for aortic stenosis.
Objective To investigate the effect of astragaloside A (AS-A) on the photoreceptor degeneration induced by sodium iodate (NaIO3) and its related mechanism. MethodsSixty healthy male C57BL/6J mice, aged 6-8 weeks, were randomly divided into normal control (NC) group, NaIO3 group, and AS-A group, with twenty mice in each group. 30 min before modeling, AS-A group mice were intraperitoneally injected with 100 μl AS-A at a dose of 100 mg/kg body weight. 30 min later, mice in NaIO3 group and AS-A group were intraperitoneally injected with 100 μl NaIO3 at a dose of 30 mg/kg body weight. Subsequently, AS-A group mice were administered AS-A twice daily at 12 h intervals until the end of the experiment. On day 1 post-modeling, zonula occludens-1 (ZO-1) immunohistochemistry was performed to observe the structure of retinal pigment epithelium (RPE) cells; real-time quantitative polymerase chain reaction (qPCR) was conducted to detect the mRNA expression of various retinal chemokine ligand-2 (Ccl2), interleukin-1 beta (Il-1β), mixed lineage kinase domain-like protein (Mlkl), receptor-interacting protein kinase 3 (Ripk3), and tumor necrosis factor (Tnf). On day 3 post-modeling, immunohistochemistry was performed to observe the expression of ionized calcium binding adaptor molecule 1 (Iba1) and glial fibrillary acid protein (GFAP) in the retina; TdT-mediated dUTP nick-end labeling (TUNEL) assay was used to detect photoreceptor cell death in each group. On day 4 post-modeling, fundus morphology of mice in each group was observed by fundus color photography and optical coherence tomography (OCT). Hematoxylin-eosin staining (HE) was used to observe the morphological structure of the retina in each group. Inter-group comparisons between two groups were conducted using independent samples t-test, while comparisons among three groups were performed using one-way ANOVA. ResultsFundus color photography and OCT examination showed that a large number of scattered yellow-white subretinal nodular structures in the fundus of NaIO3 group mice, and a large number of strong reflection areas in the RPE layer. The number of strong reflection areas in the RPE layer was reduced in the AS-A group. Immunohistochemical analysis of ZO-1 showed that ZO-1 was largely lost on the RPE cell membrane in that NaIO3 group; whereas in the AS-A group, ZO-1 was evenly distributed on the RPE cell membrane. HE staining results showed circular black deposits were visible in the RPE layer of the NaIO3 group, and the inner and outer segments of photoreceptors were severely damaged, with a significant decrease in the number of outer nuclear layer (ONL) cell nuclei; whereas in the AS-A group, the RPE layer pigments were orderly, the inner and outer segments of photoreceptors were intact, and the number of ONL cell nuclei significantly increased. The results of TUNEL staining show that numerous TUNEL-positive cell nuclei were observed in the ONL of the retina in the NaIO3 group, while the number of TUNEL-positive cell nuclei in the ONL of the retina was significantly reduced in the AS-A group, with statistically significant differences (t=2.66, P＜0.05). The analysis of qPCR data showed that compared with the AS-A group, the relative expression levels of Mlkl, Ripk3, Ccl2, Il-1β and Tnf mRNA in the retina were significantly increased in the NaIO3 group, with statistically significant differences (F=39.18, 10.66, 53.51, 41.40, 24.13; P＜0.001). Immunohistochemical staining results showed that compared with NC group and AS-A group, the positive expression of GFAP in retina of NaIO3 group was significantly increased, and the difference was statistically significant (F=9.62, P＜0.05). ConclusionAS-A antagonizes NaIO3-induced photoreceptor degeneration in part by inhibiting photoreceptor cell death and neuroinflammation. Meanwhile, AS-A treatment protects against NaIO3-triggered perturbation of retinal homeostasis.
Objective
To clarify incidence and risk factors of hepatitis B reactivation during short term (one month) in hepatitis B virus (HBV) related hepatocellular carcinoma (HCC) patients receiving partial hepatectomy.
Methods
From January 2015 to December 2015, 214 consecutive patients with HBV-related HCC who underwent partial hepatectomy were retrospectively enrolled in this study. The risk factors affecting incidence of hepatitis B reactivation were analyzed.
Results
Hepatitis B reactivation happened in 7.0% (15/214) of patients within 1 month after partial hepatectomy. By univariate analysis, the preoperative HBV-DNA negativity and hepatitis B e antigen (HBeAg) positivity were significantly correlated with the occurrence of hepatitis B reactivation (P=0.023 and P=0.001, respectively). By multivariate analysis, the preoperative HBV-DNA negativity 〔OR=9.21, 95% CI (2.40, 35.45), P=0.001〕 and HBeAg positivity 〔OR=20.51, 95% CI (5.41, 77.73), P<0.001〕 were the independent risk factors for hepatitis B reactivation.
Conclusions
Hepatitis B reactivation is common after partial hepatectomy for HBV-related HCC during short term, especially in patients whose preoperative HBV-DNA negativity and HBeAg positivity. A close monitoring of HBV-DNA during short term after partial hepatectomy is necessary, once hepatitis B is reactivated, antiviral therapy should be given.
Objective To compare the screening ability of modified Hodge test (MHT), modified carbapenem inactivation method (mCIM) and EDTA-carbapenem inactivation method (eCIM) for drug resistance phenotype of carbapenemase-producing Klebsiella pneumoniae. MethodsCarbapenem resistant Klebsiella pneumoniae (CRKP) strains clinically isolated from 5 hospitals in Chengdu between January 2019 and December 2021 were collected, and the carbapenem sensitive Klebsiella pneumoniae (CSKP) strains isolated in the same period were randomly collected. Polymerase chain reaction (PCR) -amplified carbapenem resistance gene as the gold standard, the consistencies between the results of the three phenotypic tests and the results of genetic testing were compared. Results A total of 160 CRKP strains and 120 CSKP strains were isolated. Among the 160 CRKP strains, carbapenem resistance genes were detected in 156 strains, including 105 strains of blaKPC-2, 41 strains of blaNDM-1, 8 strains of blaKPC-19, 1 strain of blaIMP-1, and 1 strain carrying both blaKPC-2 and blaNDM-1. None of the 120 CSKP drug resistance genes were detected. The sensitivity and specificity of carbapenem screening for MHT and mCIM were 73.08% (114/156), 96.67% (116/120), 97.44% (152/156) and 98.33% (118/120), respectively. The sensitivity and specificity of eCIM for screening metalloenzymes were 95.35% (41/43) and 100% (120/120), respectively. Conclusions The sensitivity of MHT to detect carbapenemase is lower than that of the other two methods, and it is easy to produce false negatives when it is used to detect metalloenzymes. The mCIM has high sensitivity and is consistent with the PCR genetic test results. The combined detection of mCIM and eCIM can screen carbapenemases more effectively and distinguish the types of carbapenemases.
ObjectiveTo observe the clinical characteristics and therapeutic effect of reactivation of retinopathy of prematurity (ROP) patients after intravitreal injection of ranibizumab (IVR). MethodsA retrospective case series study. Eleven children with ROP (21 eyes) who were reactivated after IVR in Shenzhen Eye Hospital from January 2019 to October 2021 were included in the study. Among them, there were 6 males (11 eyes) and 5 females (10 eyes), with the gestational age of (27.6±2.2) weeks and birth weight of (1 034.6±306.5) g. At the first IVR treatment, 14 eyes (63.7%, 14/22) had acute ROP (AROP), 8 eyes (36.3%, 8/22) had threshold lesions. Post-reactivation treatments include IVR, retinal laser photocoagulation (LP), or minimally invasive vitrectomy (MIVS). The follow-up time after treatment was 12 to 18 months. Birth gestational age, birth weight, treatment method, corrected gestational age at treatment, lesion stage before and after treatment, lesion reactivation and regression time were recorded. The clinical characteristics and efficacy were observed and analyzed. ResultsThe time from initial IVR treatment to reactivation was (8.2±3.5) weeks. The corrected gestational age of the child was (43.62±4.08) weeks. In 21 eyes, AROP, threshold lesion, pre-threshold lesion, and stage 4 lesion were in 2, 4, 12, and 3 eyes, respectively. The patients were treated with IVR, LP, IVR+LP, IVR+MIVS in 2, 13, 4 and 2 eyes, respectively. After the first reactivation treatment, the time of regression and stability was (8.4±4.9) weeks after treatment. There were 5 eyes with secondary reactivation of the lesion, and the lesion stages were stage 3, stage 4a and stage 5 in 2, 1 and 2 eyes, respectively. The mean reactivation time was (19.3±6.0) weeks after the last treatment. The patients in stage 3, stage 4a and stage 5 were treated with LP, LP+MIVS and IVR, respecitively, and the lesions subsided steadily during follow-up. At the last follow-up, 19 out of 21 eyes showed complete regression of the lesions, stable photocoagulation, regression of crista-like lesions, no additional lesions, and retinal leveling. All retinal detachment was "funnel-shaped" in 2 eyes. ConclusionsThe lesion reactivation of AROP after IVR treatment is more common. The early reactivation rate is higher after treatment. There is a possibility of reactivation twice after re-treatment.
Objective
To review research progress of pancreas stellate cells in pancreas fibrosis and understand characteristics and activation of pancreas stellate cells and its mechanism on pancreas fibrosis.
Method
The relevant literatures about pancreas stellate cells and its studies in pancreas fibrosis were reviewed.
Results
The activation of pancreatic stellate cell is associated with fibrosis of pancreatitis and end stage of pancreas transplantation, but its effect and regulation mechanisms for the extracelluar and intracellular molecular network need to be further investigated.
Conclusion
Elucidation of activation of pancreas stellate cells will facilitate understanding of pancreas fibrosis and searching new target in treatment of pancreas fibrosis.
【Abstract】 Objective To discuss the role of leukocyte activation and inflammatory processes in the disease of chronic venous insufficiency （CVI）. Methods The relevant literatures about the role of leukocyte activation and inflammatory reaction in CVI were reviewed. Results The role of inflammatory reaction in occurrence and development of venous diseases has been studied a lot in recent years. It was found that the leukocyte activation and inflammatory reaction are involved in the structural remodeling of venous valves and walls, leading to valvular incompetence and formation of varicose veins. Conclusion Leukocyte activation and inflammatory processes take important roles in the occurrence and progression of CVI.
ObjectiveTo investigate the imaging characteristics of gallium-68 labeled fibroblast activation protein inhibitor (68Ga-FAPI)-positron emission tomography/magnetic resonance (PET/MR) imaging in patients with liver fibrosis or liver tumor. MethodsThirteen patients with suspected liver tumor who underwent 68Ga-FAPI-PET/MR examination from May 2020 to April 2021 were retrospectively analyzed. Maximum standard uptake value (SUVmax) was investigated. All patients underwent liver surgery or biopsy. Scheuer scoring system was used to evaluate the liver fibrosis. The imaging characteristics of liver fibrosis or liver tumor were analyzed. ResultsThe liver fibrosis was confirmed in 6 patients, including 1 case of S2, 2 cases of S3, and 3 cases of S4. Among them, 4 patients had increased uptake of 68Ga-FAPI, with patchy or diffuse abnormal concentration of liver, and the SUVmax was 7.9±3.1. The liver imaging of the other 2 patients with liver fibrosis showed no obvious radioactive concentration. In addition, 2 patients were diagnosed with hepatocellular carcinoma, its SUVmax was 7.2 and 6.1; 1 patient was diagnosed with hepatobiliary duct carcinoma and its SUVmax was 13.8. Moreover, increased uptake of 68Ga-FAPI was observed in 4 patients with metastatic liver cancer, with SUVmax of 6.7±2.7. ConclusionBoth liver fibrosis and liver tumor are suitable for 68Ga-FAPI-PET/MR examination, which have different imaging characteristics.
ObjectiveTo analyze the cost and performance of five methods for detection of carbapenemase-producing Enterobacteriaceae (CPE), including PCR (method A), Carba NP test (method B), ultraviolet spectrophotometry (method C), modified carbapenem inactivation method (mCIM, method D), and loop-mediated isothermal amplification (LAMP, method E).MethodsPubMed, EMbase, The Cochrane Library, CNKI, WanFang Data and CBM databases were searched using the computer regarding literature on detection of CPE with the same or similar designs, same objectives, and independent results. The search was limited between May 2009 and May 2019. Data on the cost and detection performance of all five methods were extracted, and the four special indexes for laboratory tests, such as sensitivity, specificity, simplicity, and rapidity in the utility were quantified as specific values; subsequently, the cost-effective analysis (CEA), cost-utility analysis (CUA), and multi-attribute utility theory (MAUT) in the detection economic analysis were used to conduct health economics evaluation of five detection methods for CPE.ResultsThe cost of methods A, B, C, D and E were 210.00 yuan, 22.00 yuan, 10.50 yuan, 6.00 yuan, and 60.00 yuan, respectively. The C/E of CEA for the above five methods were 210.00, 22.96, 10.66, 6.14, and 60.00, respectively. The C/U of CUA for the above five methods were 302.16, 32.13, 19.30, 11.13, and 80.00, respectively. The MAUT value of the above five methods were 42.56, 5.00, 2.54, 1.63, and 12.56, respectively.ConclusionIn terms of CEA, CUA, and MAUT, the method D was the highest in economic value, which usually can be used as a routine method for detecting CPE, but it needs a long procedure time; thus, the method E can be used for rapid detection when clinical severe infection occurred, which is superior in both cost-effectiveness and rapidity.
Platelets are rapidly activated by activators and produce a large number of platelet microparticles (PMPs) with high coagulation activity, resulting in coagulation dysfunction. However, the generation mechanism of PMPs is still not clear. Hopping probe ion conductance microscopy (HPICM) has special technical advantages in non-contact, real-time, high-resolution imaging of living cells under physiological conditions. Using HPICM, this study monitored the processes of platelet activation and generation of PMPs in real time in the presence of calcium ionophore A23187 and cytochalasin D (CD), respectively. The results proved that the intracellular calcium concentration and the cytoskeletal proteins played important roles in the platelet activation and the generation of PMPs. Compared with the low density spread shape platelets (LDSS), the high density bubble shape platelets (HDBS) were more sensitive to the calcium ionophore A23187 and cytochalasin D. This research has a guiding significance for the further study on the relationship between platelet activation and coagulation function using HPICM.
