Cyclin-dependent kinase 9 (CDK9) is a member of the transcription CDK subfamily and plays a role in transcriptional regulation. Selective CDK9 degraders possess potent clinical advantages over reversible CDK9 inhibitors. Herein, we report the first ATG101-recruiting selective CDK9 degrader, AZ-9, based on the hydrophobic tag kinesin degradation technology. AZ-9 showed significant degradation effects and selectivity toward other homologous cell cycle CDKs in vitro and in vivo, which could also affect downstream related phenotypes. Mechanism research revealed that AZ-9 recruits ATG101 to initiate the autophagy-lysosome pathway, and forms autophagosomes through the recruitment of LC3, which then fuses with lysosomes to degrade CDK9 and the partner protein Cyclin T1. These dates validated the existence of non-proteasomal degradation pathway of hydrophobic driven protein degradation strategy for the first time, which might provide research ideas for chemical induction intervention on other types of pathogenic proteins.

[This corrects the article DOI: 10.1016/j.apsb.2021.08.008.].

Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

Critical care medicine-related treatment is an interdisciplinary and multi-professional process,often leading to secondary or concomitant mental disorders in clinical practice.Currently,there is no consensus on the pharmacological treatment of related mental illnesses in China.The Chinese Society of Psychosomatic Medicine collaborated with the Critical Care Medicine expert group to form a consensus writing expert group.After a systematic review of relevant literature,summarizing published domestic and foreign literature,and extensive discussions,the consensus was developed.The consensus elaborates on the principles and processes of the standardized use of psychotropic drugs in critical care medicine,as well as the clinical indications,precautions,and specific drug selection of various psychiatric medications,providing feasible suggestions and guidance for the clinical application of psychiatric medications in the intensive care unit.

Objective To observe the daily bladder and bowel preparation of patients with prostate cancer by cone-beam computed tomography (CBCT), and analyze its impact on the precise implementation of radiotherapy for prostate cancer and side effects. Methods We retrospectively analyzed 38 patients with prostate cancer who underwent volumetric modulated arc therapy. The number of radiation fractions for each patient ranged from 25 to 35. A CBCT scan was performed before each daily radiation therapy, and the number of scans for each patient ranged from 25 to 40. Setup errors were adjusted to ensure that the tumor was targeted and the rectum wall was not in the high-dose target area of the prostate. There were 93 instances where treatment could not be successfully implemented and re-preparation and re-scanning were required. We calculated the success rate of treatment and setup errors, compared radiotherapist-adjusted error values under different bladder and bowel preparation conditions, and recorded radiotherapy-related side effects. Results The success rate of treatment in the 38 patients was (92.14 ± 5.25)%. Among the 93 instances of seriously inadequate preparation, 48.4% were due to insufficient bladder filling, and 30.1% were due to intestinal bloating. Radiotherapy side effects were negatively correlated with the success rate of treatment (r = −0.393, P = 0.015). When bladder filling was sufficient, there were no significant differences in radiotherapist-adjusted error values in the left-right (LR), superior-inferior (SI), and anterior-posterior (AP) directions between adequate and inadequate bowel preparation (P > 0.05). When the bladder was moderately or insufficiently filled, there were significant differences in radiotherapist-adjusted error values in the LR, SI, and AP directions between adequate and inadequate bowel preparation (P < 0.05). Conclusion Insufficient bladder filling and intestinal bloating are the main factors influencing the successful implementation of radiotherapy for prostate cancer. When the bladder is sufficiently filled, bowel preparation does not affect prostate position change.

Substantial progress in the use of chemo-photodynamic nano-drug delivery systems (nano-DDS) for the treatment of the malignant breast cancer has been achieved. The inability to customize precise nanostructures, however, has limited the therapeutic efficacy of the prepared nano-DDS to date. Here, we report a structurally defined tandem-responsive chemo-photosensitive co-nanoassembly to eliminate primary breast tumor and prevent lung metastasis. This both-in-one co-nanoassembly is prepared by assembling a biocompatible photosensitive derivative (pheophorbide-diphenylalanine peptide, PPA-DA) with a hypoxia-activated camptothecin (CPT) prodrug [(4-nitrophenyl) formate camptothecin, N-CPT]. According to computational simulations, the co-assembly nanostructure is not the classical core-shell type, but consists of many small microphase regions. Upon exposure to a 660 nm laser, PPA-DA induce high levels of ROS production to effectively achieve the apoptosis of normoxic cancer cells. Subsequently, the hypoxia-activated N-CPT and CPT spatially penetrate deep into the hypoxic region of the tumor and suppress hypoxia-induced tumor metastasis. Benefiting from the rational design of the chemo-photodynamic both-in-one nano-DDS, these nanomedicines exhibit a promising potential in the inhibition of difficult-to-treat breast tumor metastasis in patients with breast cancer.

Methyltransferase like 13 (METTL13), a kind of methyltransferase, is implicated in protein binding and synthesis. The upregulation of METTL13 has been reported in a variety of tumors. However, little was known about its potential function in head and neck squamous cell carcinoma (HNSCC) so far. In this study, we found that METTL13 was significantly upregulated in HNSCC at both mRNA and protein level. Increased METTL13 was negatively associated with clinical prognosis. And METTL13 markedly affected HNSCC cellular phenotypes in vivo and vitro. Further mechanism study revealed that METTL13 could regulate EMT signaling pathway by mediating enhancing translation efficiency of Snail, the key transcription factor in EMT, hence regulating the progression of EMT. Furthermore, Snail was verified to mediate METTL13-induced HNSCC cell malignant phenotypes. Altogether, our study had revealed the oncogenic role of METTL13 in HNSCC, and provided a potential therapeutic strategy.
Head and Neck Neoplasms ; Humans ; Squamous Cell Carcinoma of Head and Neck/genetics*

Objective:To establish a disease risk prediction model for the newborn screening system of inherited metabolic diseases by artificial intelligence technology.Methods:This was a retrospectively study. Newborn screening data ( n=5 907 547) from February 2010 to May 2019 from 31 hospitals in China and verified data ( n=3 028) from 34 hospitals of the same period were collected to establish the artificial intelligence model for the prediction of inherited metabolic diseases in neonates. The validity of the artificial intelligence disease risk prediction model was verified by 360 814 newborns ' screening data from January 2018 to September 2018 through a single-blind experiment. The effectiveness of the artificial intelligence disease risk prediction model was verified by comparing the detection rate of clinically confirmed cases, the positive rate of initial screening and the positive predictive value between the clinicians and the artificial intelligence prediction model of inherited metabolic diseases. Results:A total of 3 665 697 newborns ' screening data were collected including 3 019 cases ' positive data to establish the 16 artificial intelligence models for 32 inherited metabolic diseases. The single-blind experiment ( n=360 814) showed that 45 clinically diagnosed infants were detected by both artificial intelligence model and clinicians. A total of 2 684 cases were positive in tandem mass spectrometry screening and 1 694 cases were with high risk in artificial intelligence prediction model of inherited metabolic diseases, with the positive rates of tandem 0.74% (2 684/360 814)and 0.46% (1 694/360 814), respectively. Compared to clinicians, the positive rate of newborns was reduced by 36.89% (990/2 684) after the application of the artificial intelligence model, and the positive predictive values of clinicians and artificial intelligence prediction model of inherited metabolic diseases were 1.68% (45/2 684) and 2.66% (45/1 694) respectively. Conclusion:An accurate, fast, and the lower false positive rate auxiliary diagnosis system for neonatal inherited metabolic diseases by artificial intelligence technology has been established, which may have an important clinical value.

Objective:To analyze the value of shear wave velocity (SWV) combined with thyroid stimulating hormone (TSH) in the diagnosis of hyperthyroidism.Methods:Thirty-five patients with hyperthyroidism who were treated and confirmed in the Fuyang Affiliated Hospital of Anhui Medical University from December 2017 to September 2019 were selected as hyperthyroidism group, and 30 cases of normal health check-up patients in the outpatient department were selected as control group. All of the patients and medical persons were checked by conventional two-dimensional ultrasound and SWV, and the SWV and serum TSH, thyrotrophin receptor antibody(TRAb), thyroglobulin antibody (TGAb), thyroid peroxidase antibody (TPOAb) expression levels of two groups were tested and compared. The correlation relationship in SWV value and serum TSH, TRAb, TGAb, TPOAb levels of hyperthyroidism patients was analyzed by Pearson methods. The receiver operating characteristic curve (ROC curve) method was used to analyze the value of SWV, serum TSH and SWV combined with serum in diagnosis of hyperthyroidism.Results:The SWV values of upper pole, middle pole and lower pole in the hyperthyroidism group had no significant differences ( P>0.05). The SWV values of upper hole, middle pole and lower pole of the left and right lobe of thyroid in the hyperthyroidism group were significantly higher than those in the control group [left lobe: (2.41 ± 0.34) m/s vs. (2.07 ± 0.28) m/s, (2.44 ± 0.39) m/s vs. (2.08 ± 0.25) m/s, (2.46 ± 0.43) m/s vs. (2.04 ± 0.30) m/s; right lobe: (2.47 ± 0.42) m/s vs.(2.01 ± 0.25) m/s, (2.41 ± 0.40) m/s vs. (1.95 ± 0.23) m/s, (2.43 ± 0.35) m/s vs. (2.06 ± 0.24) m/s] ( P<0.01). The serum TSH level in the hyperthyroidism group were significantly lower than that in the control group [(0.05 ± 0.03) kU/L vs. (2.74 ± 1.17) kU/L], while serum TRAb, TGAb and TPOAb levels were significantly higher than those in the control group [(15.82 ± 5.54) U/L vs. (0.55 ± 0.13) U/L, (290.63 ± 145.03) kU/L vs. (25.63 ± 7.12) kU/L, (627.17 ± 250.33) kU/L vs. (34.32 ± 5.95) kU/L], and the differences were statistically significant ( P<0.01). In the hyperthyroidism group, the SWV was negatively correlated with serum TSH ( r=- 0.862, P<0.05), but positively correlated with serum TRAb, TGAb and TPOAb ( r=0.763, 0.837, 0.804, P<0.05). The area under curve(AUC), sensitivity and specificity of combined diagnosis of hyperthyroidism with SWV value and serum TSH were 0.936, 94.29% and 91.43%, which was better than that of SWV (0.803, 80.00%, 74.29%) and serum TSH (0.842, 82.86%, 77.14%). Conclusions:SWV combined with TSH has a high clinical value in the diagnosis of hyperthyroidism.