Inhibition of KIR2.1 decreases pulmonary artery smooth muscle cell proliferation and migration.

The investigation of effective therapeutic drugs for pulmonary hypertension (PH) is critical. KIR2.1 plays crucial roles in regulating cell proliferation and migration, and vascular remodeling. However, researchers have not yet clearly determined whether KIR2.1 participates in the proliferation and migration of pulmonary artery smooth muscle cells (PASMCs) and its role in pulmonary vascular remodeling (PVR) also remains elusive. The present study aimed to examine whether KIR2.1 alters PASMC proliferation and migration, and participates in PVR, as well as to explore its mechanisms of action. For the in vivo experiment, a PH model was established by intraperitoneally injecting Sprague­Dawley rats monocrotaline (MCT). Hematoxylin and eosin staining revealed evidence of PVR in the rats with PH. Immunofluorescence staining and western blot analysis revealed increased levels of the KIR2.1, osteopontin (OPN) and proliferating cell nuclear antigen (PCNA) proteins in pulmonary blood vessels and lung tissues following exposure to MCT, and the TGF­ß1/SMAD2/3 signaling pathway was activated. For the in vitro experiments, the KIR2.1 inhibitor, ML133, or the TGF­ß1/SMAD2/3 signaling pathway blocker, SB431542, were used to pre­treat human PASMCs (HPASMCs) for 24 h, and the cells were then treated with platelet­derived growth factor (PDGF)­BB for 24 h. Scratch and Transwell assays revealed that PDGF­BB promoted cell proliferation and migration. Immunofluorescence staining and western blot analysis demonstrated that PDGF­BB upregulated OPN and PCNA expression, and activated the TGF­ß1/SMAD2/3 signaling pathway. ML133 reversed the proliferation and migration induced by PDGF­BB, inhibited the expression of OPN and PCNA, inhibited the TGF­ß1/SMAD2/3 signaling pathway, and reduced the proliferation and migration of HPASMCs. SB431542 pre­treatment also reduced cell proliferation and migration; however, it did not affect KIR2.1 expression. On the whole, the results of the present study demonstrate that KIR2.1 regulates the TGF­ß1/SMAD2/3 signaling pathway and the expression of OPN and PCNA proteins, thereby regulating the proliferation and migration of PASMCs and participating in PVR.

Whole Transcriptome Analysis Identifies the Taxonomic Status of a New Chinese Native Cattle Breed and Reveals Genes Related to Body Size.

Wandong (WD) cattle has recently been identified as a new Chinese native cattle breed by the National Commission for Livestock and Poultry Genetic Resources. The population size of this breed is less than 10,000. WD cattle and Dabieshan (DB) cattle are sympatric but are raised in different ecological environments, on mountains and plains, respectively, and the body sizes of these two breeds are markedly different. Blood samples were obtained from 8 adult female WD cattle and 7 adult female DB cattle (24 months old). The total RNA was extracted from leukocyte cells, and sequencing experiments were conducted on the Illumina HiSeqTM 4000 platform. After the removal of one outlier sample from the WD cattle breed as determined by principal component analysis (PCA), phylogenetic and population structure analyses indicated that WD and DB cattle formed a distinct Central China cattle group and showed evidence of hybridization between Bos. taurus and Bos. indicus. The immune-regulator CD48 (P = 1.3E-6) was associated with breed-specific traits according to loss-of-function variant enrichment analysis. In addition, 113 differentially expressed genes were identified between the two breeds, many of which are associated with the regulation of body growth, which is the major difference between the two breeds. This study showed that WD cattle belong to the group of hybrids between Bos. Taurus and Bos. indicus, and one novel gene associated with breed traits and multiple differentially expressed genes between these two closely related breeds was identified. The results provide insights into the genetic mechanisms that underlie economically important traits, such as body size, in cattle.

Correction: N-Primary-amine tetrapeptide-catalyzed highly asymmetric Michael addition of aliphatic aldehydes to maleimides.

Correction for 'N-Primary-amine tetrapeptide-catalyzed highly asymmetric Michael addition of aliphatic aldehydes to maleimides' by Zhi-Hong Du et al., Org. Biomol. Chem., 2020, 18, 6899-6904, DOI: 10.1039/D0OB01457E.

N-Primary-amine tetrapeptide-catalyzed highly asymmetric Michael addition of aliphatic aldehydes to maleimides.

The highly asymmetric Michael addition reaction between maleimides and aliphatic aldehydes catalyzed by low-loading ß-turn tetrapeptides with excellent yields and enantioselectivities at room temperature was reported. α-Branched and α-unbranched aldehydes both are suitable nucleophiles. N-Aryl, alkyl and hydrogen maleimides all are well tolerated and led to high yields and enantioselectivities. The transformation can be enlarged to the gram scale without decrease in the yield and enantioselectivity. Furthermore, the succinimides were converted into γ-lactams and γ-lactones, showing good practicality of this work. Some reaction intermediates in the proposed reaction mechanism can be captured with the HR-MS method.

Transcriptional activity of FIGLA, NEUROG2, and EGR1 transcription factors associated with polymorphisms in the proximal regulatory region of GPR54 gene in cattle.

Activity of transcription factors affect synthesis of G-protein coupled receptor 54 (GPR54), an important factor in regulation of initiation of puberty. Expression of the GPR54 gene in cattle is associated with polymorphisms in the proximal regulatory region (PRR) of the GPR54 gene. Transcription resulting in production of GPR54 mRNA transcript occurs as a result of transcription factor (TF) interactions in the PRR. Polymorphisms in the PRR may be associated with extent of activity of these TFs. Folliculogenesis-specific BHLH TF (FIGLA), neurogenin 2 (NEUROG2), and early growth response 1 (EGR1) are important in modulation of ovarian follicle development and neurons synthesizing GnRH, thus, regulating biosynthesis of luteinizing hormone. The aim of this study, therefore, was to assess the transcription-activating potential of binding sites for FIGLA, NEUROG2, and EGR1 TFs in the GPR54 promoter of cattle. Two luciferase-based promoters, ATC and CCT, which contain three single nucleotide polymorphisms (SNPs), A/C-794, T/C-663, and C/T-601, in the GPR54 PRR, were analyzed to evaluate gene expression and activation of different promoters by FIGLA, NEUROG2, and EGR1. The FIGLA induced GPR54 transcription through the CCT, whereas NEUROG2 and EGR1 induced GPR54 transcription through the ATC promoter-binding site. The CCT-activating effects of FIGLA were greater (2.56-fold) than the ATC-activating effects (P < 0.05). The ATC-activating effects of NEUROG2 and EGR1 were markedly greater (12.91- and 8.41-fold; P < 0.01) than CCT-activating effects. The polymorphisms, CCT and ATC, of the cattle GPR54 affect the activity of transcription factors, therefore, have an important effect on production of GPR54 mRNA transcript.

Peptide-Catalyzed Highly Asymmetric Cross-Aldol Reaction of Aldehydes to Biomimetically Synthesize 1,4-Dicarbonyls.

ß-Turn tetrapeptides were demonstrated to catalyze asymmetric aldol reaction of α-branched aldehydes and α-carbonyl aldehydes, i.e. glyoxylates and α-ketoaldehydes, to biomimetically synthesize acyclic all-carbon quaternary center-bearing 1,4-dicarbonyls in high yield and excellent enantioselectivity under mild conditions. The spatially restricted environment of the tetrapeptide warrants high enantioselectivity and yield with broad substrates. Using this protocol, (R)-pantolactone, the key intermediate of vitamin B5, was readily accessed in a practical, efficient, and environmentally benign process from inexpensive starting materials.

Double-edge Role of B Cells in Tumor Immunity: Potential Molecular Mechanism.

B cells are a heterogeneous population, which have distinct functions of antigen presentation, activating T cells, and secreting antibodies, cytokines as well as protease. It is supposed that the balance among these B cells subpopulation (resting B cells, activated B cells, Bregs, and other differentiated B cells) will determine the ultimate role of B cells in tumor immunity. There has been increasing evidence supporting opposite roles of B cells in tumor immunity, though there are no general acceptable phenotypes for them. Recent years, a new designated subset of B cells identified as Bregs has emerged from immunosuppressive and/or regulatory functions in tumor immune responses. Therefore, transferring activated B cells would be possible to become a promising strategy against tumor via conquering the immunosuppressive status of B cells in future. Understanding the potential mechanism of double-edge role of B cells will help researchers utilize activated B cells to improve their anti-tumor response. Moreover, the molecular pathways related to B cell differentiation are involved in its tumor-promoting effect, such as NF-κB, STAT3, BTK. So, we review the molecular and signaling pathway mechanisms of B cells involved in both tumor-promoting and tumor-suppressive immunity, in order to help researchers optimize B cells to fight cancer better.

Fibrin glue-assisted for the treatment of corneal perforations using glycerin-cryopreserved corneal tissue.

AIM: To evaluate the outcomes and safety of lamellar keratoplasty (LK) assisted by fibrin glue in corneal perforations. METHODS: Six eyes of 6 patients affected by different corneal pathologies (2 posttraumatic corneal scar and 3 bacterial keratitis) underwent LK procedures by using fibrin glue. The mean corneal perforation diameter was 1.35±0.64mm (range, 0.7-2.5mm), and the greatest diameter of the ulcerative stromal defect was 2.47±0.77mm in average (range, 1.5-3.5mm). The donor corneal lamella diameters were 0.20-mm larger and thicker than the recipient to restore a physiologic corneal thickness and shape: mean donor diameter was 8.34±0.28mm (range, 8.2-8.7mm) and mean thickness was 352±40.27mm (range, 220-400mm). Mean follow-up was 7.33±1.97 months (range, 6-11 months). Postoperatively, the graft status, graft clarity, anterior chamber response, the visual prognosis, intraocular pressures, and postoperative complications were recorded. RESULTS: All the corneal perforations were successfully healed after the procedure. The best-corrected visual acuity (BCVA) ranged from 20/1 000 to 20/50 in their initial presentation, and from 20/100 to 20/20 in their last visit, showed increase in all the patients. No major complications such as graft dislocation and graft failure were noted. Neovascularization developed in the superficial stroma of donor graft in 1 case. High intraocular pressure developed on day 2 after surgery, while was remained in normal range after application of anti-glaucomatous eyedrops for 1 week in 1 case. CONCLUSION: Fibrin glue-assisted sutureless LK is valuable for maintaining the ocular integrity in the treatment of corneal perforations.

Modulation of TGFß(2) and dopamine by PKC in retinal Müller cells of guinea pig myopic eye.

AIM: To investigate the effect of protein kinase C (PKC) on transforming growth factor-ß(2) (TGFß(2)) and dopamine in retinal Müller cells of guinea pig myopic eye. METHODS: Myopia was induced by translucent goggles in guinea pig, whose retinal Müller cells were cultured using the enzyme-digesting method. Retinal Müller cells were divided into 5 groups: normal control, myopia, myopia plus GF109203X, myopia plus PMA, myopia plus DMSO. PKC activities were detected by the non-radioactive methods. TGFß(2) and tyrosine hydroxylase (TH) proteins were analyzed by Western Blotting in retinal Müller cells. Dopamine was determined by the high-performance liquid chromatography-electrochemical detection in suspensions. RESULTS: After 14 days deprived, the occluded eyes became myopic with ocular axle elongating. Müller cells of guinea pigs were obtained using enzyme digestion. Compared with normal control group, the increase in PKC activity and the up-regulation in TGFß(2) expression were found in retinal Müller cells of myopic eyes, with the decrease of TH and dopamine content (P<0.05). After PKC activated by PMA, TGFß(2) and TH content were up-regulated with the increase of dopamine content (P<0.05). While the PKC activities was inhibited by GF109203X, proteins of TGFß(2) and TH were down-regulated in the myopic eyes, with the decrease of dopamine content (P<0.05). CONCLUSION: TGFß(2) and dopamine are modulated by PKC in Müller cells of the myopic eyes in guinea pig.

[Dosimetric analysis of radiotherapy with middle shielding blocks of different widths at the lower cervical supraclavicular field for stage N2-3 nasopharyngeal carcinoma].

BACKGROUND & OBJECTIVE: Anterior tangential field irradiation with middle shielding block at the lower cervical and supraclavicular region is needed in the conventional radiotherapy for stage N2-3 nasopharyngeal carcinoma (NPC), but there are still some disagreements on block width. This study was to explore a reasonable block width by dosimetric analysis of anterior tangential field irradiation with middle shielding blocks of different widths designed by the 3-dimensional treatment planning system (3D-TPS) at the lower cervical supraclavicular region for stage N2-3 NPC. METHODS: Ten untreated patients with stage N2-3 NPC received 3D-TPS-designed irradiation plan. For every patient, a gradual shrinking field technique was adopted. Single anterior tangential fields were set at the lower cervical and supraclavicular region and irradiated with middle shielding blocks of different widths: 0 cm (Plan A), 2.1 cm (Plan B), 2.5 cm (Plan C), and 3 cm (Plan D) for the first 40 Gy, then 3 cm for residual dose for all 4 plans. The prescribed doses were the same for 4 plans for every patient. The irradiated volumes and doses of target volumes and organs at risk among the 4 plans were compared. RESULTS: The high dose coverage (V95 and V90) of plan target volume (PTV) for the subclinical lesion region at the lower cervical supraclavicular region (PTV50a) was significantly higher in Plan A than in Plans B, C, and D (82.44% vs. 78.21%, 77.10% and 73.80% for V95, 87.89% vs. 84.03%, 82.68% and 77.50% for V90, P<0.05), and significantly higher in Plans B and C than in Plan D (P<0.05), but there was no difference between Plans B and C (P>0.05). There was no significant difference in high dose coverage (V95 and V90) of PTV for the primary gross tumor region (PTVnx), PTV for the cervical metastatic nodes (PTVnd), PTV for the high risk region around primary gross tumor (PTVnx60), PTV for the high risk region around metastatic nodes (PTVnd60), and subclinical lesion region above cricoid cartilage (PTV50b) among the 4 plans. There was no difference in the doses for the spinal cord and larynx among the 4 plans. The maximal doses for 50% volumes of target organs (D50) were significantly higher in Plan A than in Plans B, C, and D (49.47 Gy vs. 41.95 Gy, 38.73 Gy, and 26.82 Gy for the thyroid gland, 44.52 Gy vs. 8.41 Gy, 7.03 Gy, and 5.63 Gy for the esophagus, 44.18 Gy vs. 10.16 Gy, 8.55 Gy, and 7.60 Gy for the trachea, P<0.05), and higher in Plans B and C than in Plan D (P<0.05), but there was no difference between Plans B and C (P>0.05). The normal tissue complication probability (NTCP) of the thyroid gland was significantly higher in Plan A than in Plans B, C, and D (7.9% vs. 4.8%, 4.3%, and 3.0%, P<0.05), and higher in Plans B and C than in Plan D, but there was no difference between Plans B and C (P>0.05). There were no difference in the doses for the spinal cord and larynx among the 4 plans. The maximal doses for 50% volumes of target organs (D50) were significantly higher in Plan A than in Plans B, C, and D (49.47 Gy vs. 41.95 Gy, 38.73 Gy, and 26.82 Gy for the thyroid gland, 44.52 Gy vs. 8.41 Gy, 7.03 Gy, and 5.63 Gy for the esophagus, 44.18 Gy vs. 10.16 Gy, 8.55 Gy, and 7.60 Gy for the trachea, P<0.05), and higher in Plans B and C than in Plan D (P<0.05), but there was no difference between Plans B and C (P>0.05). The normal tissue complication probability (NTCP) of the thyroid gland was significantly higher in Plan A than in Plans B, C, and D (7.9% vs. 4.8%, 4.3%, and 3.0%, P<0.05), and higher in Plans B and C than in Plan D, but there was no difference between Plans B and C (P>0.05). There were no difference in the NTCP of other organs at risk among the 4 plans (P>0.05). CONCLUSIONS: Not obviously increasing the irradiation doses for critical organs at risk, Plan A has the best high dose coverage at the subclinical lesion region of the lower cervical and supraclavicular region, while Plan D the worst. We recommend to use no middle shielding block in the anterior tangential field for the first 40 Gy, then use individual middle shielding blocks of 2.1-2.5 cm in the institutes at where set up error is small.

[Sparing normal oral tissues with individual dental stent in radiotherapy for primary nasopharyngeal carcinoma patients].

BACKGROUND & OBJECTIVE: With the progression of radiotherapy techniques, the 5-year overall survival rate of nasopharyngeal carcinoma (NPC) patients has increased obviously. As the survival time prolonged, more and more attention was paid to various radiation sequelae and the quality of life of the patients. This study was to explore the role of individual dental stent in sparing normal oral tissues for primary NPC patients in radiotherapy by pushing the tongue and a part of oral mucous membrane away from the radiation fields. METHODS: Irradiation dose and volume of the tongue of a NPC patient before and after wearing dental stent was evaluated. A total of 43 patients were randomized into 2 groups: 19 in trial group and 24 in control group. Trial group wore dental stent during radiotherapy, while control group did not. Patients' weight, taste, oral mucous reaction, and tongue mucous reaction before radiotherapy and every week during radiotherapy were examined. RESULTS: Dosimetric analysis proved that the irradiation dose and volume of the tongue decreased obviously in trial group. The occurrence rate of grade 1-2 mucositis of the oral cavity was higher in trial group than in control group (73.68% vs. 62.50%), but the occurrence rate of grade 3-4 mucositis was lower in trial group than in control group (26.32% vs. 37.50%, P=0.470). By the completion of radiotherapy, 4 (21.05%) patients in trial group and 19 (79.17%) in control group suffered from taste dysfunction (P<0.001). CONCLUSION: Individual dental stent is useful in sparing the oral mucous membrane and preserving taste for primary NPC patients in radiotherapy.