A Ca2+ sensor BraCBL1.2 involves in BraCRa-mediated clubroot resistance in Chinese cabbage.

Clubroot disease caused by Plasmodiophora brassicae (P. brassicae) severely threatens the cultivation of Cruciferous plants, especially Chinese cabbage. Recently, resistance genes in plants have been reported to encode for a Ca2+-permeable channel in the plasma membrane, which can mediate the cytosolic Ca2+ increase in plant cells upon pathogen attack. However, the downstream Ca2+ sensor and decoder are still unknown. In this study, we identified the virulent and avirulent P. brassicae isolates (Pbs) of two near isogenic lines, CR 3-2 and CS 3-2, with CR 3-2 harboring clubroot resistant gene BraCRa. The transcriptomic analysis was then conducted with CR 3-2 after inoculating with virulent isolate PbE and avirulent isolate Pb4. From the differentially expressed genes of transcriptomic data, we identified a Ca2+-sensor encoding gene, BraCBL1.2, that was highly induced in CR 3-2 during infection by Pb4 but not by PbE. Moreover, GUS histochemical staining and subcellular localization analysis revealed that BraCBL1.2 was specifically expressed in the root hair cells of Arabidopsis and encoded a putative Ca2+ sensor localized in the plasma membrane. We also developed an assay to investigate the BraCRa-mediated hypersensitive response (HR) in tobacco leaves. The results suggest that BraCBL1.2 is involved in the BraCRa-mediated plant ETI immune response against P. brassicae. In addition, we verified that overexpression of BraCBL1.2 enhanced clubroot resistance in Arabidopsis. Collectively, our data identified the involvement of a Ca2+ sensor in BraCRa-mediated clubroot resistance in Chinese cabbage, providing a theoretical basis for further research on the resistance of Chinese cabbage to P. brassicae.

Application and the Effect of the Triple Prerehabilitation Nursing Model in the Perioperative Period of Knee Arthroplasty in Diabetic Patients.

Objective: The aim of the study was to explore the application and the effect of the triple prerehabilitation nursing model in the perioperative period of knee arthroplasty in diabetic patients. Methods: The prospectively included 60 patients with diabetes who underwent total knee replacement were admitted from August 2021 to April 2022 and were divided into 2 groups according to the (1 : 1) ratio. The control group was mainly given routine nursing care. On the basis of the control group, the observation group received triple prerehabilitation nursing. The postoperative knee flexion, hospital for the special surgery knee score (HSS), the daily living ability (Barthel) score, the modified fall efficacy scale (MFES) score, the recovery of the lower-limb muscle strength, and the incidence of complications were compared between the two groups. Results: The knee flexion degree and lower-limb muscle recovery of the observation group were better than those of the control group at 3 d, 7 d, and 14 d after operation (P < 0.05). The HSS score, Barthel score, and MFES score of the observation group were higher than those of the control group (P < 0.05). There was no significant difference in postoperative complications between the two groups (P > 0.05). Conclusion: The triple prerehabilitation nursing care for diabetic patients undergoing total knee replacement can promote the recovery of limb function.

Characterization of a pathogenic variant in GBA for Parkinson's disease with mild cognitive impairment patients.

Parkinson's disease (PD) is the second most common neurodegenerative disease, and mild cognitive impairment (MCI) is a well-established risk factor for the development of dementia in PD. A growing body of evidence suggests that low expression of glucocerebrosidase (GBA) promotes the transmission of α-synuclein (α-Syn) interpolymers and the progression of PD. However, how GBA mutations affect the pathogenesis of PD via abnormal aggregation of α-Syn is unclear, and no clinically valid PD-MCI genetic markers have been identified. Here, we first located a GBA eQTL, rs12411216, by analysing DHS, eQTL SNP, and transcription factor binding site data using the UCSC database. Subsequently, we found that rs12411216 was significantly associated with PD-MCI (P < 0.05) in 306 PD patients by genotyping. In exploring the relationship between rs12411216 and GBA expression, the SNP was found to be associated with GBA expression in 50 PD patients through qPCR verification. In a further CRISPR/Cas9-mediated genome editing module, the SNP was identified to cause a decrease in GBA expression, weaken enzymatic activity and enhance the abnormal aggregation of α-Syn in SH-SY5Y cells. Additionally, using an electrophoretic mobility shift assay, we confirmed that the binding efficiency of transcription factor E2F4 was affected by the rs12411216 SNP. In conclusion, our results showed that rs12411216 regulated GBA expression, supporting its potential role as a PD-MCI genetic biomarker and highlighting novel mechanisms underlying Parkinson's disease.

CT pulmonary angiography using different noise index values with an iterative reconstruction algorithm and dual energy CT imaging using different body mass indices: Image quality and radiation dose.

OBJECTIVE: To investigate the differences in imaging quality and radiation dose in CT pulmonary angiography (CTPA) by using fast-kV switching dual energy CT imaging and 3D Smart mA modulation at different body mass indices (BMIs) and at different noise index (NI) values with an adaptive statistical iterative reconstruction (ASIR) algorithm. METHODS: Four hundred patients who underwent CTPA were equally divided into two groups: A (18.5 kg/m2 ≦ BMI <24.9 kg/m2) and B (24.9 kg/m2 ≦ BMI ≦ 4.9 kg/m2). The groups were randomly subdivided into four subgroups (n = 50): A1-A4 and B1-B4. The patients in subgroups A1 and B1 underwent fast-kV switching dual energy CT imaging. The other patients underwent 3D Smart mA modulation with the ASIR algorithm at NI values 26, 36, and 46 for A2/B2, A3/B3, and A4/B4, respectively. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) of all images were calculated after CTPA. Images were then subjectively evaluated using a 5-point scale. The volume CT dose index and dose-length product (DLP) were recorded and their means calculated. The DLP was converted to the effective dose (ED). RESULTS: In group A, the SNR, CNR, and subjective image scores showed no statistical differences (P > 0.05). The ED in subgroup A4 was 67.12% and 31.53% lower than that in A1 and A2, respectively. In group B, the variables showed no significant differences between the subgroups B3, B1, and B2 (P > 0.05). The ED in subgroup B3 was 50.12% and 35.95% lower than that in B1 and B2, respectively. CONCLUSIONS: Setting different NI values according to BMIs and applying the ASIR algorithm can more effectively reduce the radiation dose in CTPA than in fast-kV switching dual energy CT, while maintaining image quality. Imaging may be performed at NI = 46 in patients with lower BMI (group A) and at NI = 36 in patients with higher BMI (group B).

Correlation between conjunctival and corneal calcification and cardiovascular calcification in patients undergoing maintenance hemodialysis.

The purpose of this study was to investigate the correlation of conjunctival and corneal calcification (CCC) with cardiovascular calcification in patients undergoing maintenance hemodialysis (MHD). A total of 122 patients undergoing MHD in our hospital were included in this study. Conjunctival and corneal calcification was examined by slit lamp and graded. Abdominal aortic calcification (AAC), aortic valve calcification (AVC), and mitral valve calcification (MVC) were determined by X-ray or ultrasound. The correlation of CCC with AAC, AVC, and MVC was analyzed. Biochemical, hematological, and cardiovascular data were compared between patients with different severity of CCC or AAC. Mitral valve calcification was significantly associated with AAC in our patients. Conjunctival and corneal calcification positively correlated with AAC. We also found that patients with severe CCC exhibited significantly higher levels of serum calcium, phosphate, product of calcium and phosphate, serum copper, cystatin, intact parathyroid hormone, and vitamin D than patients with mild CCC. In addition to significantly increased levels of serum calcium, product of calcium and phosphate, serum copper, and cystatin, patients with severe AAC also had higher high-sensitivity C-reactive protein level and greater left ventricular posterior wall thickness and left ventricular end-diastolic interventricular septum thickness than patients with mild AAC. Our results suggest that patients undergoing MHD with severe CCC or AAC have high degree of mineral metabolism disorder, inflammation, and cardiovascular function disorder. The strong correlation between CCC and AAC indicates that CCC score might be used as an indirect indicator to predict cardiovascular risks in patients undergoing MHD.

ROS-Ca(2+) is associated with mitochondria permeability transition pore involved in surfactin-induced MCF-7 cells apoptosis.

The surfactin can inhibit proliferation and induce apoptosis in cancer cells. Moreover, surfactin can induce cell death in human breast cancer MCF-7 cells through mitochondrial pathway. However, the molecular mechanism involved in this pathway remains to be elucidated. Here, the reactive oxygen species (ROS) and Ca(2+) on mitochondria permeability transition pore (MPTP) activity, and MCF-7 cell apoptosis which induced by surfactin were investigated. It is found that surfactin evoked mitochondrial ROS generation, and the surfactin-induced cell death was prevented by N-acetylcysteine (NAC, an inhibitor of ROS). An increasing cytoplasmic Ca(2+) concentration was detected in surfactin-induced MCF-7 apoptosis, which was inhibited by 1,2-bis (2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA-AM, a chelator of calcium). In addition, the relationship between ROS generation and the increase of cytoplasm Ca(2+) was determined. The results showed that surfactin initially induced the ROS formation, leading to the MPTP opening accompanied with the collapse of mitochondrial membrane potential (ΔΨ(m)). Then the cytoplasmic Ca(2+) concentration increased in virtue of the changes of mitochondrial permeability, which was prevented by BAPTA-AM. Besides, cytochrome c (cyt c) was released from mitochondria to cytoplasm through the MPTP and activated caspase-9, eventually induced apoptosis. In summary, surfactin has notable anti-tumor effect on MCF-7 cells, however, there was no obvious cytotoxicity on normal cells.

Up regulation of annexin A2 on murine H22 hepatocarcinoma cells induced by cartilage polysaccharide.

In this study, we investigated the antitumor effect of a tumor vaccine prepared from H22 hepatocarcinoma cells induced by cartilage polysaccharide. We found out there were specific antigens which combined with antigen-specific antibodies from immune murine serum. Results of western blot analysis showed that about 36 kDa make specific antibodies appeared specific antibodies in antiserum of immune mice, whereas the best immune effects became visible at the induction time of 48 h. Analyses of 2-dimensional electrophoresis identified the specific antigen was annexin A2, which was a glycosylated protein that contained a glycosylation site, closely related to oncogenesis, cancer development, invasion and metastasis. Proteomics indicated that both quantity and conformation of annexin A2 were changed after induced by cartilage polysaccharide. Lastly, we found there was a major increase of annexin A2 mRNA on H22 cells induced by cartilage polysaccharide. In summary, our data suggested that annexin A2, a specific antigen played a key role in antitumor immune response and activating the immune system. It would be a potential type of tumor vaccine which provided new ideas for tumor immunoprophylaxis.

[Effects of surfactin on proliferation, apoptosis and cytoskeleton in human breast cancer MCF-7 cells].

We studied the effect of surfactin on cell proliferation, apoptosis and the cytoskeleton in human breast cancer cell line MCF-7 in vitro. The result of 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) showed that the surfactin inhibited proliferation of MCF-7 cells in a dose- and time-dependent manner, with IC50 at 48 h of 27.3 micromol/L. Surfactin-induced cell death was considered to be apoptotic by observing the typical apoptotic morphological changes by AO/EB staining. Flow cytometric analysis also demonstrated that surfactin caused time-dependent apoptosis of MCF-7 cells through cell arrest at G2/M phase. Immunofluorescence and Western blotting showed that surfactin significantly suppressed the expression of vimentin, induced the alpha-tubulin depolymerization and rearrangement and then the skeleton system of the cells changed dramatically. Based on our findings, surfactin can significantly inhibit the growth of MCF-7 cells and induce apoptosis.