The circadian clock plays a critical role in regulating various physiological processes, including gene expression, metabolic regulation, immune response, and the sleep-wake cycle in living organisms. Post-translational modifications (PTMs) are crucial regulatory mechanisms to maintain the precise oscillation of the circadian clock. By modulating the stability, activity, cell localization and protein-protein interactions of core clock proteins, PTMs enable these proteins to respond dynamically to environmental and intracellular changes, thereby sustaining the periodic oscillations of the circadian clock. Different types of PTMs exert their effects through distincting molecular mechanisms, collectively ensuring the proper function of the circadian system. This review systematically summarized several major types of PTMs, including phosphorylation, acetylation, ubiquitination, SUMOylation and oxidative modification, and overviewed their roles in regulating the core clock proteins and the associated pathways, with the goals of providing a theoretical foundation for the deeper understanding of clock mechanisms and the treatment of diseases associated with circadian disruption.
Protein Processing, Post-Translational/physiology* ; Circadian Rhythm/physiology* ; Humans ; Animals ; CLOCK Proteins/physiology* ; Circadian Clocks/physiology* ; Phosphorylation ; Acetylation ; Ubiquitination ; Sumoylation

As one of the chronic diseases with high incidence in contemporary society, cervical spondylosis has increasing patient groups who gradually present a low age, and it seriously affects social and public health. Although modern medicine has made great progress in the pathological research and clinical treatment of cervical spondylosis, patients still face gastrointestinal side effects of nonsteroidal anti-inflammatory drugs(NSAIDs), neck pain, limited mobility, upper limb numbness, and other symptoms after conservative or surgical treatment. In the theory of traditional Chinese medicine(TCM), cervical spondylosis belongs to the categories of "Bi syndrome" "stiff neck" "stiff Bi", etc. With the change of the times, the change of lifestyle, and the application of western medicine treatment, the etiology and pathogenesis of TCM in cervical spondylosis also show new characteristics. In terms of etiology and pathogenesis, it involves the invasion of wind, cold, and dampness, long-term strain, liver and kidney deficiency, Qi and blood stasis, which are associated with factors such as cervical degeneration, muscle tension and spasm, intervertebral disc herniation, and nerve root compression in modern medicine. In terms of the evolution of pathogenesis, in the early stage, wind, cold, and dampness, were more common in Xuanfu, resulting in unfavorable muscles and bones, poor flow of Qi and blood, and cervical spondylosis and radiculopathy. Medium-term phlegm stasis and internal knots, sluggish muscles and veins, and long-term weathering and fire are more likely to occur in the vertebral artery and sympathetic radiculopathy. In the later stage, the positive Qi is depleted; the true Yin is damaged, and the viscera Qi and blood are deficient, which is most common in cervical myelopathy. The strategy of treating cervical spondylosis with TCM classic formulas applies Gegen Decoction, Wutou Decoction, Qianghuo Shengshi Decoction, Mahuang Jiazhu Decoction to patients with wind, cold, and dampness. Patients with phlegm dampness and blood stasis are treated with Huoxue Xiaoling Dan, Jinlingzi Powder, Siwu Decoction, Banxia Baizhu Tianma Decoction, Shuanghe Decoction, etc. For those patients with liver, spleen, and kidney deficiency, Huangqi Guizhi Wuwu Decoction, Tianma Gouteng Decoction, Guishao Dihuang Pills, Shenling Baizhu Powder, and Lizhong Decoction are used to invigorate the spleen, nourish Qi and blood, and tonify liver and kidney. In clinical practice, the authors advocate a safe and effective treatment plan of classic formulas based on deficiency and excess, the integration of formulas and syndromes, and the combination of modern research results, so as to relieve symptoms, reduce recurrence, and reduce medical burden.
Humans ; Spondylosis/drug therapy* ; Medicine, Chinese Traditional/methods* ; Drugs, Chinese Herbal/therapeutic use* ; Cervical Vertebrae/pathology*

Objective To analyze the disposition of medical support equipment support units for medical service support of the synthetic battalion to carry out efficiently demand matching and replenishment for war loss,so as to enhance the efficiency of deployment and decision making of medical support equipment.Methods War loss of the medical support equipment was analyzed with queuing theory with the support service of the synthetic battalion as the basic research framework and desired equipment intact rate in the battlefield as the goal.A M/M/1/∞/∞ model and a M/M/c/∞/∞ model were established to investigate the disposition process of medical support equipment support units by simulating actual conditions of the battlefield.Multi synthetic battalions were assumed to be deployed to carry out 2 missions with different tasks and combat intensities,of which 40 synthetic battalions were involved in for mission 1 and 35 synthetic battalions for mission 2.The disposition of medical support equipment support units was calculated with the two models constructed.Results With the M/M/c/∞/∞ model it's suggested 5 medical support equipment support units be deployed to serve for 40 synthetic battalions;with the M/M/1/∞/∞ model only one unit was employed to support 35 battalions.The two models both met the requirements of 90％intact rate.Conclusion The research contributes to the decision making of medical support equipment support forces during synthetic battalion medical support and enhances the precision support of medical support equipment.[Chinese Medical Equipment Journal,2024,45(4):83-87]

Objective To evaluate the effect of delayed cleaning on the cleaning and disinfection quality of gastro-scopes after pre-treatment with different solutions.Methods According to the factorial design table,combination of the pre-treatment cleaning solutions(factor A)(including multi-enzyme cleaning solution[A1],clean water[A2])and delayed cleaning durations(factor B)(including 0 minutes after pre-treatment[B1],30 minutes after pre-treat-ment[B2],1 hour after pre-treatment[B3],and 3 hours after pre-treatment[B4])yielded eight groups(A1B1,A1B2,A1B3,A1B4,A2B1,A2B2,A2B3,A2B4).According to the usage order of gastroscopes,96 gastroscopes used in the digestive endoscopy center of a tertiary first-class hospital from May to September,2023 were randomly assigned to each group by random number table method,with 12 gastroscopes in each group.Specimens were taken at four time points:after pre-treatment,before cleaning,after cleaning,and after disinfection.Due to instant clea-ning,no specimen before cleaning were taken from A1B1 and A2B1 groups,thus only 3 specimens were taken from these two groups each.Four specimens were taken from gastroscopes in the rest groups,resulting in 360 specimens in total.The internal condition of the biopsy cavity was observed through a cavity detector during each delayed cleaning period after pre-treatment,and specimens were taken at the subsequent reprocessing processes of the gas-troscopes.The microbial conditions of the gastroscopes after pre-treatment,before cleaning,after cleaning,and af-ter disinfection were compared.Results After pre-treatment with multi-enzyme cleaning solution and clean water,there was no statistically significant difference in microbiological detection result(P＞0.05).The biopsy cavity re-mained moist during the delayed cleaning period.There was no statistically significant difference in the microbial de-tection results of factors A and B before and after delayed cleaning as well as after disinfection(all P＞0.05).There was no interaction effect between factor A and B.The distribution of bacterial colonies and disinfection qualified rate of gastroscopes after pre-treatment with two cleaning solutions were also not statistically different(both P＞0.05).Conclusion Delayed cleaning for 30 minutes,1 hour,and 3 hours after pre-treatment does not affect the cleaning and disinfection quality of gastroscopes.When clinical demand is urgent,immediate cleaning should be carried out.However,a certain buffering time(no longer than 3 hours)before cleaning is acceptable,when cleaning and disin-fection workload is heavy and timely cleaning cannot be carried out.

Objective To observe the residual of lumbago and leg pain with contained type(CT)and non-contained type(NCT)lumbar disc herniation(LDH)after transforaminal endoscopic treatment,and to explore the role of hypoxia-inducible factor-1α(HIF-1α)and transient receptor potential vanillate 1(TRPV1)pathway.Methods A total of 68 single-segment LDH patients were selected from July 2021 to October 2022,including 44 males and 24 females;aged 26 to 67 years old with an av-erage of(43.63±11.94)years old;course of disease was 4 to 36(18.91±10.34)months;body mass index was(24.45±4.00)kg·m-2;there were 7 cases of L3.4 segments,32 cases of L4,5 segments,and 29 cases of L5S1 segments.All of them were per-formed with percutaneous intervertebral endoscopic extraction of nucleus pulposus and were divided into contained group(CT group)and non-contained group(NCT group)with 34 cases respectively according to the integrity of outer layer of fibrous an-nulus observed during operation.A total of 17 patients who underwent open surgery for scoliosis or vertebral fracture were se-lected as control group,including 12 males and 5 females;aged 21 to 65 years old with an average of(39.41±12.80)years old;body mass index was(24.86±4.11)kg·m-2.The relative mRNA expression quantity of HIF-1α,TRPV1 in nucleus pulposus were measured by quantitative real-time PCR.The contents of neurokinin 1 receptor(NK1R),nerve growth factor(NGF),vascular endothelial growth factor(VEGF)in nucleus pulposus and the serum substance P(SP)and calcitonin gene-related peptide(CGRP)were detected by enzyme linked immunosorbent assay(ELISA).The threshold of lumbar tenderness was de-tected by a pressure pain meter.The degree of lumbago and lumbar function were evaluated by visual analog scale(VAS)and Oswestry disability index(ODI)separately.The residual rate of postoperative lumbago and leg pain was assessed.Results The mRNA relative expression quantity of HIF-1α and TRPV1,and the contents of NK1R,NGF and VEGF in nucleus pulposus,and the levels of serum SP and CGRP before surgery in the NCT group were higher than those in the CT group(P＜0.05),and those in the CT group were higher than the control group(P＜0.05).At day 7 after surgery,the serum SP and CGRP levels,lum-bago and leg pain VAS scores and lumbar ODI index in two LDH groups were lower than before surgery(P＜0.05),and those in the NCT group were higher than the CT group(P＜0.05),and the threshold of lumbar tenderness in the NCT group was lower than the CT group(P＜0.05).The differences of lumbago and leg pain VAS scores,lumbar ODI index and lumbar tenderness threshold between preoperative and postoperative 7 days in the NCT group were lower than those in the CT group(P＜0.05).The residual rate of lumbago and leg pain at 7 days after surgery in the NCT group was higher than that in the CT group(P＜0.05).Conclusion HIF-1α and TRPV1 pathway promoted the excessive production of NGF,VEGF,NK1R in nucleus pulposus and serum neuropeptides SP and CGRP,which may lead to the higher residual rate of lumbago and leg pain with non-contained lumbar disc herniation postoperative.

This work was aimed at predicting and verifying B-cell epitopes of SARS-CoV-2 E protein through bioinformatics methods,and clarifying the dominant B cell epitopes with mouse polyclonal antibody serum prepared through SARS-CoV-2 re-combinant E protein immunization and human positive serum vaccinated with inactivated SARS-CoV-2 vaccine.The structural and B-cell linear epitopes of SARS-CoV-2 E protein were predicted with SOPMA,Expasy,SWISS-MODEL,IEDB database,and Bepid-2.0 software.Candidate epitopes were expressed as GST-tagged recombinant protein fragments in an E.coli sys-tem,and their immunoreactivity with mouse and human poly-clonal positive serum against SARS-CoV-2 E protein was de-tected by western blotting and indirect ELISA,respectively.The epitope prediction results showed that E protein contained linear B cell epitopes Ser6-Val14 and Tyr57-Pro71,and the conformational epitopes of Glu8-Val12,Leu39-Tyr59,and Ser60-Leu65.The GST tagged recombinant E protein fragments of E1 and E3,containing Ser6-Val14 and Tyr57-Pro71 epitopes,respectively,as well as E2 without an epitope sequence as a control,were expressed in an E.coli expression system and successfully purified with an Ni-NTA column.Western blotting and indirect ELISA analysis indicated that all mouse and human SARS-CoV-2 positive sera positively reacted with only E1 and E3 proteins,but negatively reacted with E2 protein,thus indicating that the corresponding epitope prediction with Ser6-Val14 and Tyr57-Pro71 was correct.This study successfully predicted and preliminarily identified two linear B cell epitopes of SARS-CoV-2 E protein,thus providing a reference for the preparation of new coronavirus vaccines and the analysis of immune response characteristics.

Objective:To investigate the efficacy and prognosis of chemotherapy regimen containing Bruton's tyrosine kinase(BTK)inhibitor in the treatment of relapsed/refractory mantle cell lymphoma(R/R MCL).Methods:The clinical data of 134 patients with R/R MCL were collected and analyzed retrospectively.The clinical characteristics of patients and effect of chemotherapy regimen on efficacy,overall survival(OS)and progression-free survival(PFS)were observed.Results:The median age of the patients was 58(56-61)years old,and male to female ratio was about 2.9∶1.Patients with Ann Arbor stage Ⅲ-Ⅳ accounted for 77.6％,extranodal involvement＞2 for 43.3％,bone marrow involvement for 60.4％,gastrointestinal involvement for 24.6％,and hepatosplenomegaly for 38.1％.The median follow-up time was 30(2-103)months,overall response rate(ORR)was 41.8％,3-year PFS was not reached,and 3-year and 5-year OS rate was 62.7％and 53.8％,respectively.The ORR of BTK inhibitor group was 56.9％,which was higher than 32.5％of non-BTK inhibitor group(P=0.006).The difference was statistically significant in PFS between the two groups(P=0.002),but was not in OS(P＞0.05).The difference was statistically significant in OS between classical and special morphology(P＜0.001),but was not in PFS(P＞0.05).Ki-67 was an influencing factor for OS and PFS.Multivariate analysis showed that Ki-67,B symptoms,MIPI score,and Ann Arbor stage were independent prognostic factors affecting patients'OS.The second-line treatment regimen was an independent prognostic factor affecting patients'PFS.Conclusions:The chemotherapy regimen containing BTK inhibitors can effectively improve the efficacy and prolong the PFS of R/R MCL patients.Ki-67,B symptoms,MIPI score,and Ann Arbor stage are independent prognostic factors for R/R MCL patients.

Aim To investigate the effect of homocys-teine(Hcy)on the permeability of pulmonary micro-vascular endothelial cells(PMVECs)and the role and mechanism of RASAL1.Methods CBS+/-mice were fed a high methionine diet(HMD)for 16 weeks to replicate an animal model of hyperhomocysteinemia(HHcy).HE staining was used to observe the changes in lung tissue structure.qRT-PCR was used to detect the levels of RASAL1 and DNMT1 mRNA in lung tis-sue.Western blot was used to detect the expression of RASAL1,DNMT1,ZO-1,and VE cadherin proteins.Methylation specific PCR was used to detect methyla-tion in the RASAL1 promoter region.PMVECs were transfected with Ad-RASAL1 to detect the expression of ZO-1 and VE cadherin.The si-DNMT1 interference fragment was transfected into PMVECs,and the ex-pression of the RASAL1 was detected by qRT-PCR and Western blot.Results Serum Hcy level of HMD mice was significantly raised,and HE staining showed severe structural disorder in lung tissue.The expres-sion of RASAL1,ZO-1,and VE cadherin was de-creased,while the expression of DNMT1 was in-creased.The degree of methylation in the RASAL1 promoter region was raised.The expression of ZO-1 and VE cadherin increased after PMVECs were trans-fected with Ad-RASAL1.After knocking down DN-MT1,RASAL1 expression was increased.Conclusion Hcy can increase the permeability of PMVECs,and its mechanism is related to the upregulation of RASAL1 methylation level.

This paper aimed to study the effect of Erjing Pills on the improvement of neuroinflammation of rats with Alzheimer's di-sease(AD) induced by the combination of D-galactose and Aβ_(25-35) and its mechanism. SD rats were randomly divided into a sham group, a model control group, a positive drug group(donepezil, 1 mg·kg~(-1)), an Erjing Pills high-dose group(9.0 g·kg~(-1)), and an Erjing Pills low-dose group(4.5 g·kg~(-1)), with 14 rats each group. To establish the rat model of AD, Erjing Pills were intragastrically administrated to rats for 5 weeks after 2 weeks of D-galactose injection. D-galactose was intraperitoneally injected into rats for 3 weeks, and then Aβ_(25-35) was injected into the bilateral hippocampus. The new object recognition test was used to evaluate the learning and memory ability of rats after 4 weeks of intragastric administration. Tissues were acquired 24 h after the last administration. The immunofluorescence method was used to detect the activation of microglia in the brain tissue of rats. The positive expressions of Aβ_(1-42) and phosphory protein Tau~(404)(p-Tau~(404)) in the CA1 area of the hippocampus were detected by immunohistochemistry. The levels of inflammatory factors interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6) in the brain tissue were determined by enzyme-linked immunosorbent assay(ELISA). Toll-like receptor 4(TLR4)/nuclear factor kappa B(NF-κB)/nucleotide-binding oligomerization domain-like receptors 3(NLRP3) pathway-associated proteins in the brain tissue were determined by Western blot. The results showed that as compared with the sham group, the new object recognition index of rats in the model control group decreased significantly, the deposition of Aβ_(1-42) and p-Tau~(404) positive protein in the hippocampus increased significantly, and the levels of microglia activation increased significantly in the dentate gyrus. The levels of IL-1β, TNF-α, and IL-6 in the hippocampus of the model control group increased significantly, and the expression levels of TLR4, p-NF-κB p65/NF-κB p65, p-IκBα/IκBα, and NLRP3 proteins in the hippocampus increased significantly. Compared with the model control group, the Erjing Pill groups enhanced the new object recognition index of rats, decreased the deposition of Aβ_(1-42) and the expression of p-Tau~(404) positive protein in the hippocampus, inhibited the activation of microglia in the dentate gyrus, reduced the levels of inflammatory factors IL-1β, TNF-α, and IL-6 in the hippocampus, and down-regulated the expression levels of TLR4, p-NF-κB P65/NF-κB P65, p-IκBα/IκBα, and NLRP3 proteins in the hippocampus. In conclusion, Erjing Pills can improve the learning and memory ability of the rat model of AD presumably by improving the activation of microglia, reducing the expression levels of neuroinflammatory factors IL-1β, TNF-α, and IL-6, inhibiting the TLR4/NF-κB/NLRP3 neuroinflammation pathway, and decreasing hippocampal deposition of Aβ and expression of p-Tau, thereby restoring the hippocampal morphological structure.
Animals ; Rats ; Rats, Sprague-Dawley ; NF-kappa B ; NF-KappaB Inhibitor alpha ; NLR Family, Pyrin Domain-Containing 3 Protein ; Galactose ; Interleukin-6 ; Neuroinflammatory Diseases ; Toll-Like Receptor 4 ; Tumor Necrosis Factor-alpha

OBJECTIVE: To screen the prognostic biomarkers of metabolic genes in patients with multiple myeloma (MM), and construct a prognostic model of metabolic genes. METHODS: The histological database related to MM patients was searched. Data from MM patients and healthy controls with complete clinical information were selected for analysis.The second generation sequencing data and clinical information of bone marrow tissue of MM patients and healthy controls were collected from human protein atlas (HPA) and multiple myeloma research foundation (MMRF) databases. The gene set of metabolism-related pathways was extracted from Molecular Signatures Database (MSigDB) by Perl language. The biomarkers related to MM metabolism were screened by difference analysis, univariate Cox risk regression analysis and LASSO regression analysis, and the risk prognostic model and Nomogram were constructed. Risk curve and survival curve were used to verify the grouping effect of the model. Gene set enrichment analysis (GSEA) was used to study the difference of biological pathway enrichment between high risk group and low risk group. Multivariate Cox risk regression analysis was used to verify the independent prognostic ability of risk score. RESULTS: A total of 8 mRNAs which were significantly related to the survival and prognosis of MM patients were obtained (P<0.01). As molecular markers, MM patients could be divided into high-risk group and low-risk group. Survival curve and risk curve showed that the overall survival time of patients in the low-risk group was significantly better than that in the high risk group (P<0.001). GSEA results showed that signal pathways related to basic metabolism, cell differentiation and cell cycle were significantly enriched in the high-risk group, while ribosome and N polysaccharide biosynthesis signaling pathway were more enriched in the low-risk group. Multivariate Cox regression analysis showed that the risk score composed of the eight metabolism-related genes could be used as an independent risk factor for the prognosis of MM patients, and receiver operating characteristic curve (ROC) showed that the molecular signatures of metabolism-related genes had the best predictive effect. CONCLUSION Metabolism-related pathways play an important role in the pathogenesis and prognosis of patients with MM. The clinical significance of the risk assessment model for patients with MM constructed based on eight metabolism-related core genes needs to be confirmed by further clinical studies.
Humans ; Cell Cycle ; Multiple Myeloma/genetics* ; Prognosis ; Risk Factors