AIM: To investigate the mechanism of Hexue Mingmu Tablets(HXMMT)in improving diabetic retinopathy(DR)based on transcriptomics.METHODS: Zebrafish DR models were established by 3-day glucose induction(130 mmol/L)starting at 3 days post-fertilization(dpf). Larvae were randomized into four groups: control group(CG; aquaculture water), model group(MG; 130 mmol/L glucose), low-dose HXMMT treatment group(L-HX; 130 mmol/L glucose +7.5 mg/L HXMMT), and high-dose HXMMT treatment group(H-HX; 130 mmol/L glucose +75 mg/L HXMMT), with a 3-day intervention period until 6 dpf. The area and length of eyes, and body length of zebrafish were observed by stereomicroscopy, retinal morphology was observed by hematoxylin-eosin staining(HE), and retinal vessel diameter was observed under fluorescence microscope. Differentially expressed genes(DEGs)were identified by RNA-sequencing(RNA-seq)technology to further elucidate the molecular mechanism of HXMMT in improving DR in zebrafish, and the sequencing accuracy was validated through quantitative real-time polymerase chain reaction(qRT-PCR).RESULTS: HE staining demonstrated that the intervention with HXMMT significantly improved the disordered cell arrangement, widened gaps, and thickened inner nuclear layer(INL)in ganglion cell layer GCL); retinal vascular diameter quantification revealed that the retinal vessel diameter of the MG significantly increased compared with the CG, and it was significantly changed after the intervention of HXMMT, with significant efficacy in the H-HX(P<0.05); transcriptomics profiling identified 1 470 reversed DEGs, predominantly enriched in the AMPK signaling pathway, FoxO signaling pathway, retinal developmental processes, and tight junction regulation. Technical validation confirmed strong correlation between qRT-PCR and RNA-seq data(R2=0.8571, P<0.05).CONCLUSION: HXMMT may improve retinal vascular microcirculation disorders in DR by regulating core targets including vsx1, pde6c, arr3a, plk1, fbp1b, foxo1a, pcna, and cdk1, as well as synergistically modulating processes such as retinal development in camera-type eyes, visual perception, microtubule cytoskeletal organization, tight junctions, and the AMPK signaling pathway, Foxo signaling pathway.

OBJECTIVE To identify core genes associated with the treatment of coronary heart disease （CHD） with Tongmai yangxin pills， and predict their potential immunological and metabolic mechanisms. METHODS Mendelian randomization （MR） analysis was conducted using protein quantitative trait loci （pQTL） data from the UK Biobank and Icelandic，and data from genome- wide association study to screen core genes related to Tongmai yangxin pills in the treatment of CHD. Gene expression changes were further validated using transcriptomic sequencing data. Mediation analyses of immune cells and plasma metabolites were subsequently performed to explore the downstream regulatory networks of these core genes. RESULTS A total of 62 positive pQTL genes showed significant causal associations with CHD. MR analysis combined with transcriptomic sequencing validation identified three core genes FAM3D，OXT， and ENPP5-associated with Tongmai yangxin pills in the treatment of CHD. The transcriptomic sequencing results showed that after treatment with Tongmai yangxin pills， the expression levels of FAM3D and OXT were significantly reduced （P＜0.01）， while the expression level of ENPP5 was significantly increased （P＜0.05）. Mediation analyses between immune cells and plasma metabolites indicated that these genes may positively or negatively regulate CHD through immune pathways involving regulatory T cells and myeloid dendritic cells expressing CD11c and CD62L， as well as through metabolic pathways related to lipid and fatty acid metabolism， cholesterol metabolism， and bile acid metabolism. CONCLUSIONS This study identified FAM3D，OXT， and ENPP5 as core genes associated with the treatment of CHD by Tongmai yangxin pills， which may exert therapeutic effects via modulation of immune cells and plasma metabolic pathways involving fatty acids and bile acids.

Objective To assess inter-observer variations(IOV)in the delineation of target volumes and organs-at-risk(OAR)for intensity-modulated radiotherapy(IMRT)of nasopharyngeal carcinoma(NPC)among physicians from different levels of cancer centers,thereby providing a reference for quality control in multi-center clinical trials.Methods Twelve patients with NPC of different TMN stages were randomly selected.Three physicians from the same municipal cancer center manually delineated the target volume(GTVnx)and OAR for each patient.The manually modified and confirmed target volume(GTVnx)and OAR delineation structures by radiotherapy experts from the regional cancer center were used as the standard delineation.The absolute volume difference ratio(△V_diff),maximum/minimum volume ratio(MMR),coefficient of variation(CV),and Dice similarity coefficient(DSC)were used to compare the differences in organ delineation among physicians from different levels of cancer centers and among the 3 physicians from the same municipal cancer center.Furthermore,the IOV of GTVnx and OAR among physicians from different levels cancer centers were compared across different TMN stages.Results Significant differences in the delineation of GTVnx were observed among physicians from different levels of cancer centers.Among the 3 physicians,the maximum values of △V_diff,MMR,and CV were 97.23%±83.45%,2.19±0.75,and 0.31±0.14,respectively,with an average DSC of less than 0.7.Additionally,there were considerable differences in the delineation of small-volume OAR such as the left and right optic nerves,chiasm,and pituitary,with average MMR>2.8,CV>0.37,and DSC<0.51.However,relatively smaller differences were observed in the delineation of large-volume OAR such as the brainstem,spinal cord,left and right eyeballs,and left and right mandible,with average△V_diff<42%,MMR<1.55,and DSC>0.7.Compared with the differences among physicians from different levels cancer centers,the differences among the 3 physicians from the municipal cancer center were slightly reduced.Furthermore,there were also differences in the delineation of target volumes for NPC among physicians from different levels cancer centers,depending on the staging of the disease.Compared with the delineation of target volumes for earlier stage patients(stages I or II),the differences among physicians in the delineation of target volumes for advanced stage patients(stages III or IV)were smaller,with average △V_diff and DSC of 98.31%±67.36%vs 69.38%±72.61%(P<0.05)and 0.55±0.08 vs 0.72±0.12(P<0.05),respectively.Conclusion There are differences in the delineation of GTVnx and OAR in radiation therapy for NPC among physicians from different levels of cancer centers,especially in the delineation of target volume(GTVnx)and small-volume OAR for early-stage patients.To ensure the accuracy of multicenter clinical trials,it is recommended to provide unified training to physicians from different levels of cancer centers and review their delineation results to reduce the effect of differences on treatment outcomes.

Objective:To analyze the early predictive value of olfactory function for cognitive decline in older adults with mild cognitive impairment (MCI) .Methods:From March 2023 to January 2024, convenience sampling was used to select 385 older adults with MCI from two community health service centers in Dongcheng District and Fengtai District of Beijing as participants. Participants were followed up for six months to collect data on general information, olfactory function testing, and cognitive function. According to whether there were changes in clinical cognitive function, older adults were divided into a cognitive function decline group and a cognitive function non-decline group. Binary Logistic regression was used to analyze the independent influencing factors of cognitive decline in older adults with MCI, and the receiver operating characteristic (ROC) curve was used to evaluate the predictive effect of olfactory function on cognitive decline.Results:Among 385 older adults with MCI, 113 (29.4%) experienced cognitive decline. Binary Logistic regression analysis showed that education level, passive cognition activity, social activity, and olfactory dysfunction were independent influencing factors for cognitive decline in older adults with MCI ( P<0.05). The area under the ROC curve was 0.819 [95% CI (0.773, 0.865), P<0.01], with an optimal cutoff value of 9.5 points, a sensitivity of 81.3% and specificity of 31.0%. Conclusions:The olfactory function has good predictive value for cognitive decline in older adults with MCI, and can be used for early screening of MCI high-risk individuals with rapid cognitive decline.

Objective To explore the construction and practical effects of a multidisciplinary collaborative nursing management model of"organization-training-emergency response-integration-quality control"for pan-vascular disease patients in integrated wards.Methods On the basis of literature research and expert meetings,a multidisciplinary collaborative nursing management model for patients with pan-vascular disease in intergrated wards has been formed.It includes the implementation of accurate dynamic and intelligent overall allocation of beds in the hospital,the formulation of multidisciplinary training programs,the establishment of an emergency and critical case treatment guarantee mechanism,the construction of a standardized path for the joint management of multidisciplinary nursing teams,and the establishment of an intelligent and homogeneous nursing quality control chain.From July 2022 to December 2023,the program was implemented in the cardio-cerebrovascular ward of a tertiary A hospital in Zhejiang province,and indicators such as the average length of stay,cure and improvement rate,and cardiopulmonary resuscitation success rate were collected,and the satisfaction of patients with pan-vascular disease was investigated.Results During the application of multidisciplinary collaborative nursing management model for patients with pan-vascular disease,5416 patients were admitted,with an average length of stay of 6.11±0.73 days,a cure and improvement rate of 98.06%(5 311/5 416),a cardiopulmonary resuscitation success rate of 94.12%,and no adverse events.The overall satisfaction score of patients improved from 92.74±1.68 points in the 4th quarter of 2022 to 98.21±0.55 points in the 4th quarter of 2023(P<0.001).Conclusion The construction of the multidisciplinary collaborative nursing management model in the integrated pan-vascular ward has explored a new path for the hospital to cultivate cross-professional nursing personnel,promoted effective resource integration,and optimized the health management path,effectively improved the utilization rate of existing resources in the hospital and patients'medical experience.

As a ligand-dependent transcription factor,retinoid-associated orphan receptor γt(RORyt)that controls T helper(Th)17 cell differentiation and interleukin(IL)-17 expression plays a critical role in the pro-gression of several inflammatory and autoimmune conditions.An emerging novel approach to the therapy of these diseases thus involves controlling the transcriptional capacity of RORyt to decrease Th17 cell development and IL-17 production.Several RORyt inhibitors including both antagonists and inverse agonists have been discovered to regulate the transcriptional activity of RORyt by binding to orthosteric-or allosteric-binding sites in the ligand-binding domain.Some of small-molecule inhibitors have entered clinical evaluations.Therefore,in current review,the role of RORyt in Th17 regulation and Th17-related inflammatory and autoimmune diseases was highlighted.Notably,the recently developed RORyt inhibitors were summarized,with an emphasis on their optimization from lead compounds,ef-ficacy,toxicity,mechanisms of action,and clinical trials.The limitations of current development in this area were also discussed to facilitate future research.

Objective:To investigate the role and mechanism of N 6-methyladenosine (m 6A) methyltransferase-like 3 (METTL3) in vascular calcification (VC) of chronic kidney disease (CKD) through apoptosis-associated protein. Methods:(1) Real-time fluorescence quantitative PCR was used to test METTL3 mRNA in serum of maintenance hemodialysis (MHD) patients. (2) Western blotting was used to detect the expression of METTL3 protein in high-phosphorus stimulated vascular smooth muscle cells (VSMCs), and immunofluorescence double lable was used to observe the distribution of METTL3 and Runt-related transcription factor 2 (Runx2). The METTL3 overexpressed and knockdown plasmids were constructed and transfected into VSMCs. Alizarin red staining was used to detect calcification degree. Western blotting was used to detect the expressions of osteogenic markers [Runx2, bone morphogenetic protein-2(BMP-2), collagen Ⅰ] and apoptosis- related proteins Bax and Bcl-2. (3) SD rats were randomly divided into control group, CKD-VC group and S-adenosylhomocysteine (SAH) intervention group. The calcification of thoracic aorta was evaluated by von Kossa staining, and the protein expressions of Runx2, Bax and Bcl-2 were detected by immunohistochemistry and Western blotting.Results:(1) METTL3 mRNA expression in MHD patients with VC was significantly lower than that in non-VC patients ( P<0.05), and was negatively correlated with coronary artery calcium score ( r=-0.65, P<0.001). (2) The expression of METTL3 in VSMCs stimulated by high phosphorus was decreased and showed a time dependence. Immunofluorescence double label showed that METTL3 and Runx2 were co-expressed in the nucleus. METTL3 was overexpressed in high-phosphorus induced VSMCs, and the expressions of Runx2, collagen I and BMP-2 were significantly decreased, accompanied by the decrease of calcified nodules and Bax/Bcl-2 ratio (all P<0.05). Conversely, METTL3 knockdown aggravated VSMCs calcification by inducing apoptosis. (3) Furthermore, METTL3 inhibitor SAH was administered in vivo, and it was found that inhibition of METTL3 expression significantly increased the calcification of rat thoracic aorta, and the Bax/Bcl-2 ratio and Runx2 expression were up-regulated. Conclusions:Serum METTL3 level is reduced in MHD patients with VC. In vivo and in vitro studies demonstrate that METTL3 inhibits VC in CKD by mediating the apoptosis-related protein Bax/Bcl-2.

L-glutamic acid is the world's largest bulk amino acid product that is widely used in the food, pharmaceutical and chemical industries. Using Corynebacterium glutamicum G01 as the starting strain, the fermentation by-product alanine content was firstly reduced by knocking out the gene encoding alanine aminotransferase (alaT), a major by-product related to alanine synthesis. Secondly, since the α-ketoglutarate node carbon flow plays an important role in glutamate synthesis, the ribosome-binding site (RBS) sequence optimization was used to reduce the activity of α-ketoglutarate dehydrogenase and enhance the glutamate anabolic flow. The endogenous conversion of α-ketoglutarate to glutamate was also enhanced by screening different glutamate dehydrogenase. Subsequently, the glutamate transporter was rationally desgined to improve the glutamate efflux capacity. Finally, the fermentation conditions of the strain constructed using the above strategy were optimized in 5 L fermenters by a gradient temperature increase combined with a batch replenishment strategy. The glutamic acid production reached (135.33±4.68) g/L, which was 41.2% higher than that of the original strain (96.53±2.32) g/L. The yield was 55.8%, which was 11.6% higher than that of the original strain (44.2%). The combined strategy improved the titer and the yield of glutamic acid, which provides a reference for the metabolic modification of glutamic acid producing strains.
Glutamic Acid ; Corynebacterium glutamicum/genetics* ; Ketoglutaric Acids ; Metabolic Engineering ; Alanine

Objective:To establish and apply the electronic further modified early warning score system (e-fMEWS), and explore its role in the condition evaluation and early warning of inpatients in non-critical units, so as to provide clinical nurses with an early and dynamic method to identify the potential deterioration risk of patients' condition.Methods:A retrospective analysis of 262 805 inpatients in multiple non-critical units of the Second Affiliated Hospital of Zhejiang University School of Medicine from January to December 2018 and January to December 2020 was performed. The patients who were hospitalized from January to December 2018 were used as the control group, and the responsible nurse used the traditional single evaluation index to start the emergency response system; the patients from January to December 2020 were used as the research group, and the emergency response system was started using e-fMEWS. The inclusion criteria were as follows: (1) hospitalization time ≥24 h; (2) patient ≥14 years old. Exclusion criteria were as follows: (1) patients had cardiopulmonary resuscitation before admission; (2) patients discontinued treatment or were transferred to another hospital during treatment; (3) patients received palliative care; (4) patients were admitted to non-critical wards in grade I of emergency pre-examination and triage. The activation of the rapid response team (RRT), the activation of the cardiorespiratory arrest team, the incidence of cardiac and respiratory arrest, the number of cases of invasive mechanical ventilation, the number of cases admitted to the intensive care unit, the length of hospital stay and the prognosis were compared. Statistical software SPSS 22.0 was used for data analysis.Results:Under the e-fMEWS assessment, compared with the control group, the rate of initiation of the research group decreased by 0.03%. For patients who initiated RRT, the average length of hospital stay was shortened, and the number of in-hospital respiratory cardiac arrest decreased (12.2% vs. 13.2%) and the number of cases transferred to the intensive care unit was less (42.8% vs. 50.6%), the rate of improvement and recovary increased (58.4% vs. 56.1%).Conclusions:The application of e-fMEWS can help clinical nurses to quickly and accurately identify the potential risk of deterioration of the patient's condition. Through early identification of potentially critically ill patients in non-critical units, early intervention and timely treatment can avoid adverse events and improve the patient prognosis.

Objective:To explore the status quo of self-stigma in diabetic patients and analyze its correlation with self-compassion.Methods:In this cross-sectional study, from December 2020 to January 2021, totally 240 diabetic patients admitted at the Department of Endocrinology in two ClassⅢ Grade A hospitals in Hangzhou, Zhejiang Province were selected by convenient sampling and investigated with the general information questionnaire, Self-Stigma Scale (SSS) , and Self-Compassion Scale (SCS) . Hierarchical regression analysis was used to explore the relationship between self-stigma and self-compassion in diabetic patients. A total of 240 questionnaires were issued, and 225 valid questionnaires were recovered. The effective response rate was 93.75% (225/240) .Results:The total SSS score of the 225 diabetic patients was (56.47±16.72) , and the average score of all items was (2.01±0.60) ; the total SCS score was (86.46±12.79) , and the average score of all items was (3.33±0.49) . Pearson correlation analysis showed that the total SSS score of diabetic patients was negatively correlated with the total SCS score ( P < 0.01) . Hierarchical regression analysis revealed that self-compassion was an influencing factor of self-stigma in diabetic patients ( P< 0.01) , which could independently explain 13.1% of self-stigma (Δ R2=0.131, P<0.001) . Conclusions:Self-stigma of diabetic patients is at a moderate level, and self-compassion is an influencing factor of self-stigma of diabetic patients. Nursing staff can reduce patients ' self-stigma by improving their self-compassion.