Background: Traditional treatment modalities for vertebral compression fractures (VCFs) include bed rest, pain medications, muscle relaxants, back braces, and physical therapy. In cases where conservative treatment proves ineffective, a new procedure called core decompression of the vertebral body is explored. Core decompression of the vertebral body has the potential to lower and stabilize the intraosseous pressure, resulting in enhanced blood circulation, which contributes to pain reduction. In this trial, we evaluated the efficacy of core decompression of the vertebral body in patients with painful VCFs compared with conventional conservative treatment. Methods: This prospective randomized controlled trial was conducted at a tertiary education hospital between June 2017 and May 2020. The participants were randomly assigned in a 1:1 ratio to one of two treatment groups: the core decompression group and the conservative treatment group. The primary outcome measure was the visual analog scale (VAS) pain score of the back 3 months after the procedure. Secondary outcome measures included the Oswestry Disability Index (ODI) for lumbar disabilities, the European Quality of Life-5 Dimensions (EQ-5D) score for quality of life, and radiographic outcomes such as changes in compression rate. Results: All patients underwent the assigned intervention (48 core decompression and 50 conservative treatments). At both 1 month and 3 months, there were no significant differences between the core decompression group and conservative treatment group in VAS pain score . In addition, there were no significant inter-group differences in ODI and EQ-5D scores throughout the follow-up period . Conclusion: Core decompression of the vertebral body did not demonstrate any significant improvement in pain and disability compared to conventional conservative treatment. Vertebral compression fractures (VCFs) can result in various complications, including chronic back pain, progressive spinal deformity, muscle wasting, pressure ulcers, and sleep disturbances ,2,3,4. AThe underlying mechanisms responsible for pain following VCFs have not been thoroughly investigated. However, increased pressure within the bone (intraosseous pressure) is considered the primary trigger ,11,12,13In this single center, randomized, controlled trial, we evaluated the efficacy of core decompression of the vertebral body in patients with painful VCFs compared with conventional conservative treatment. We hypothesized that patients who undergo core decompression of the vertebral body would report lesser pain and pain-related disabilities than those in the control group.ClinicalTrials.gov (NCT02902250). Before enrollment, written informed consent was obtained from each patient or their legal representatives. The trial was conducted in accordance with the principles of the Declaration of Helsinki. The clinical and radiographic data were monitored by an independent researcher.The participants were enrolled at a tertiary education hospital between June 2017 and May 2020. This trial was approved by the institutional review board of our hospital (B-1502-288-005) and registered at The participants were recruited from the outpatient department of our hospital by a single specialist. The inclusion criteria were as follows: (1) aged ≥ 40 years and voluntarily agreed to participate and (2) diagnosed with one to three painful osteoporotic VCFs between vertebral levels T10 and L5. In all patients with suspected compression fractures, lumbar X-rays and MRI were performed to confirm the fracture and bone marrow edema. Also, in all patients diagnosed with VCFs, bone density scans were performed to confirm osteoporotic VCFs. The exclusion criteria were as follows: (1) previous diagnosis of an inflammatory disease or malignant tumor, (2) presence of a bleeding or coagulation disorder, (3) use of any type of anticoagulant, (4) severe pain other than in the lower back, and (5) deemed unfit for participation in this clinical trial by the researcher.Participants who met the inclusion criteria were randomized into one of two treatment groups at a 1:1 ratio: the core decompression group and the conservative treatment group. The time from fracture to randomization was less than one month. Web-based randomization was employed to achieve concealed allocation. All participants were evaluated for the efficacy and safety of the treatment over a 3-month study period.At baseline, patients scored their back pain using the visual analog scale  and completed self-reported questionnaires, such as the Oswestry Disability Index (ODI) and the 5-level version of the European Quality of Life-5 (EQ-5D). The ODI is a widely used questionnaire designed to assess the level of disability or functional impairment experienced by individuals with low back pain. The EQ-5D score is a standardized instrument used to measure health-related quality of life. It assesses an individual’s general health and well-being and is commonly used in healthcare and clinical research to evaluate the overall health status and quality of life of patients. They also provided their demographic and clinical information. Evaluation measures were performed before randomization and at various times for up to 3 months. The primary outcome measure was the VAS pain score of the back. Secondary outcome measures included ODI and EQ-5D scores.The compression rate and wedge angle of the vertebral body were measured on plain radiographs . The heiAt the first visit, written informed consent, demographic information, medical history, medications, smoking status, and vital signs were obtained, and bone density scan, VAS, ODI, EQ-5D questionnaires, and radiologic examinations were conducted. On day 1 (visit 2), vital signs, VAS, ODI, and adverse event monitoring were performed. Visit 3 (at week 1) included vital signs, VAS, ODI, and adverse event monitoring, The fourth and fifth visits, at weeks 4 and 12, involved vital signs, VAS, ODI, EQ-5D, X-ray, and adverse event monitoring. Adverse events (AEs), vital signs, and physical examination were assessed. An adverse event is defined as a compression rate of 80% or more or a VAS pain score of 8 or higher.The practitioner of core decompression in this trial was an experienced orthopedic surgeon . Patients in the core decompression group were placed in a prone position. The fractured vertebrae were identified and marked under C-arm guidance. Under local anesthesia, a Jamshidi needle was used to perforate the fractured vertebrae. The needle was carefully inserted through the skin and soft tissues layer-by-layer with continuous monitoring through the pedicle using C-arm guidance. The core wire was extracted from the needle and a syringe was connected to the needle. Subsequently, the plunger of the syringe was gradually withdrawn, resulting in the evacuation of blood and intraosseous decompression. Subsequently, the needle was removed and an aseptic dressing was applied to the puncture site.In the conservative treatment group, the patients received a regimen that included bed rest, oral pain medication, use of external braces, and ambulation based on their pain tolerance.In both groups, pain management followed a stepladder approach starting with non-narcotics, low-dose narcotics, and high-dose narcotics. Pain treatment was categorized following the World Health Organization analgesic ladder as 0 (no prescription), 1 , 2 (low-dose narcotics), and 3 (high-dose narcotics). Esomeprazole, a proton pump inhibitor, was used for medication-induced dyspepsia.The degree of effectiveness of core decompression performed in this trial has not been previously reported. Therefore, the required sample size was calculated assuming that an effect similar to that of VP performed in general VCFs can be expected. Considering a minimal clinically significant difference of 1.5 on the VAS, a standard deviation of 2.5 in both groups , and a tracking loss rate of 0.1, a sample size of 49 individuals per group is required.For the primary analyses, we used an intention-to-treat strategy, and treatment effects and confidence intervals were calculated using analysis of covariance (ANCOVA) models with adjustment for baseline values of the outcome measure and age. In addition, we used analysis of variance for repeated measures (RM-ANOVA) to compare the ODI score, EQ-5D score, compression rate, and wedge angle. SPSS  was used for all statistical analyses.From June 2017 to May 2020, a total of 98 patients were enrolled and underwent randomization. Of the 98 patients, 48 and 50 were assigned to the core decompression and conservative treatment group, respectively. Both groups had six patients each who were lost to follow-up before the 3-month assessment . Demograp = 0.970). At 3 months, the mean VAS score in the core decompression group was 29.2 ± 17.0, as compared with 31.1 ± 17.1 in the conservative treatment group  (p < 0.001).The study groups did not differ significantly with respect to the primary outcome (VAS score) at 1 month and 3 months. At 1 month, the mean VAS score in the core decompression group was 43.0 ± 16.8, as compared with 43.4 ± 17.6 in the conservative treatment group  . Both grp = 0.917 and p = 0.704, respectively). This indicates that the ODI and EQ-5D scores did not differ significantly between the core decompression and conservative treatment groups throughout the follow-up period.p = 0.560). Similar results were observed for the wedge angle. At baseline, the wedge angle in the core decompression and conservative treatment groups were 12.2 ± 7.5 and 13.0 ± 7.1, respectively. After 3 months, the wedge angles in the core decompression and conservative treatment groups were 15.8 ± 9.0 and 14.0 ± 7.6, respectively. There were no significant differences between the study groups (p = 0.753).Radiological outcomes are summarized in No serious adverse events, resulting in life-threatening conditions or severe sequelae, were observed.This randomized controlled trial aimed to compare the effectiveness and safety of a newly designed vertebral body core decompression technique with those of conventional conservative treatment in patients with painful VCFs. The results indicated that the core decompression group did not demonstrate a significant difference in pain improvement compared with the conservative treatment group. Moreover, there were no significant differences in the measures of disability and quality of life between the two groups. Additionally, no significant differences were observed in the radiological parameters of the study groups at baseline and 3 months.The core decompression technique employed in this study was based on the principles of the widely used core decompression technique for ONFH . PreviouA few studies have conducted procedures similar to the core decompression performed in this study, referred to as vertebral body perforation, and have reported favorable results. Ogihara et al. demonstrated that vertebral body perforation provides sufficient pain relief in patients with painful lumbar compression fractures . They atThis study has several limitations. First, we were unable to compare the intraosseous pressure of patients in the conservative treatment and core decompression groups, despite recognizing the importance of intraosseous pressure in the pain reduction effect of core decompression. Second, patients’ knowledge of their treatment assignments may have influenced their responses to the questionnaire. Third, the follow-up period was limited to 3 months. Given that pain and disability resulting from vertebral compression fractures can affect a patient’s quality of life over an extended period, a short follow-up duration may not fully capture the long-term effects.In this prospective randomized controlled trial involving patients with osteoporotic VCF, we found no beneficial effects in performing core decompression of the vertebral body compared with that of conservative treatment in patients with painful VCFs.


The peptide sequence information is transformed into useful taxonomical information that allows to obtain the different biomass contributions at different taxonomical ranks. This new methodology is applied for the first time to analyze the composition of biofilms from minute quantities of material collected from a pool used to store radioactive sources in a nuclear facility. For these biofilms, we report the identification of three genera, namely Sphingomonas, Caulobacter, and Acidovorax, and their functional characterization by metaproteomics which shows that these organisms are metabolic active. Differential expression of Gene Ontology GOslim terms between the two main microorganisms highlights their metabolic specialization.Sampling small amounts of biofilm from harsh environments such as the biofilm present on the walls of a radioactive material storage pool offers few analytical options if taxonomic characterization and estimation of the different biomass contributions are the objectives. Although 16S/18S rRNA amplification on extracted DNA and sequencing is the most widely applied method, its reliability in terms of quantitation has been questioned as yields can be species-dependent. Here, we propose a tandem-mass spectrometry proteotyping approach consisting of acquiring peptide data and interpreting then against a generalist database without any  Deinococcus deserti isolated from the Sahara desert  and dose rate received on the walls were estimated taking into account water column attenuation.2, for less than 1 min, the pole was slowly raised, the tape removed and the sponge placed in a sterile hermetic plastic bag for transport. In the analytical laboratory, each sponge containing the biofilm was introduced inside a sterile flask containing 100 ml of sterile water. To suspend the biofilm, the sponge was stirred with a sterile spatula and the flask was shaken vigorously to detach the biological material from the sponge fibers. After removing the sponge, the resulting liquid was centrifuged at 15,000 g for 20 min at room temperature to pellet the biological material. All but 1 ml of the supernatant was discarded and the pellet was dissolved in this 1 ml volume. For metaproteomics analysis, a 750 μl volume of the suspension was stored at −20°C until metaproteomic analysis.Biofilms were sampled directly from the pool walls at different locations (specified in the Results section) using sterile Spontex scraping sponges (Spontex-Mappa). For each sample, a sponge was taped to a metal pole and was then used to scrub the wall underwater. After scrubbing an area of approximatively 100 cmg for 10 min. The resulting pellet was dissolved in 25 μl of LDS 1× Laemmli buffer (Invitrogen) consisting of 106 mM Tris/HCl, 141 mM Tris base, 2% lithium dodecyl sulfate, 10% glycerol, 0.51 mM EDTA, 0.22 mM SERVA Blue G250, 0.175 mM phenol red, buffered to pH 8.5 and supplemented with 2.5% beta-mercaptoethanol. The sample was heated at 99°C for 5 min, followed by treatment in an ultrasonic bath for 5 min. The sample was then loaded onto a 4–12% gradient 10-well NuPAGE gel (Invitrogen). Proteins were subjected to electrophoresis for 5 min as previously described  containing 200 mg of glass beads as recommended . The samescribed . After sescribed .3CN in presence of 0.1% formic acid. The gradient during MS/MS acquisition consisted in 50 min from 2 to 20% CH3CN, followed by 10 min from 20 to 32% CH3CN. A quick wash of the column was done with an additional gradient from 32 to 72% CH3CN in 1 min, before equilibrating the column with 2% CH3CN. The Q-Exactive HF instrument was operated with a Top20 strategy: full scan mass spectra were acquired from m/z 350 to 1,500 with an Automatic Gain Control Target and resolution set at 3 × 106 ions and 60,000, respectively, for the MS scan, and 1 × 105 ions and 15,000, respectively, for the MS/MS scan. MS/MS scan acquisition was started when the intensity threshold of 170,000 was reached. The maximum fill time was 60 ms and the isolation window for the quadrupole was 1.6 m/z. Only ions with potential charge states of 2+ and 3+ were selected for MS/MS with a dynamic exclusion time of 10 s.The extracted peptides (10 μl) were analyzed with a Q-Exactive HF mass spectrometer (Thermo) equipped with an ultra-high field Orbitrap analyser and coupled to an Ultimate 3000 176 RSL Nano LC System (ThermoFisher). This analysis was performed in data-dependent mode in similar conditions as those previously described , i.e., f1 Peptide-Spectrum Matches (PSMs) were validated with a Mascot expectation value below 0.05 using Mascot identity threshold (MIT), and by allowing multiple PSMs per MS/MS spectrum for ion scores higher than 98% of the top ion score. Peptides associated to spectra were assigned to taxa, resulting in Taxon-Spectrum Matches (TSMs) at each possible taxonomic level, species, genus, family, order, class, phylum, and superkingdom, as previously defined  contributes to the selection of high-confidence taxa, at the expense of sensitivity.At each taxonomical rank, the pertaining taxa were ranked by the number of specific peptides, from highest to lowest. The top taxon per parent was compared to the specific peptides threshold, cycling through parent taxa. If a validated parent had no validated subtaxon, the child with the maximum number of TSMs was validated as the best representative of the clade. TSMs corresponding to validated taxa were added to a pool of TSMs at the taxonomical rank . This was reiterated for the next taxa in order, one per parent, until no more taxa with sufficient specific peptides could be found. The subtaxa were then reordered by the number of corresponding TSMs, excluding RankValTSMs, and compared to the threshold TSM, cycling through the parent taxa. The process was ended when the number of RankValTSMs (rank) was above 0.992 × RankValTSMs (parents), or when no additional taxon could meet validation criteria. The conjunction of a Mascot MIT Sphingomonas, Caulobacter, and Acidovorax contained in the NCBInr fasta file, as well as the most common proteomics contaminants (MaxQuant contaminants.fasta database). The Sphingomonas-Caulobacter-Acidovorax pan-proteomic database comprises 498,167 protein sequence entries totaling 163,630,882 amino acids from 4,032 different annotated genomes. Peptide-to-MS/MS spectrum matches (PSMs) with a MASCOT expectation p-value below 0.002612 were selected. This p-value threshold was obtained by searching the merged data set on the forward and reverse database as a decoy, and adjusting the p-value to correspond to a false discovery rate of 1%. Proteins were grouped on the basis of their shared peptides. Protein abundances were assessed by their spectral counts. Gene Ontology (GO) annotations of proteins were obtained using DIAMOND  and 1,136 at the upper level, i.e., 4% of the TSMs are shared between both classes. Fractions based on specific peptides would estimate Betaproteobacteria at 55/580 = 9.5% of the bacterial content, while their quantification based on TSMs would be 171/1136 = 15%, which is more accurate in the case of low peptide overlap between clades. At the Order rank, the Betaproteobacteria taxon was explained by the presence of a unique order: Burkholderiales with 47 unique peptides and 160 TSMs. The Alphaproteobacteria taxon signal is subdivided into two children orders: Sphingomonadales and Caulobacterales. More taxonomic precision could be achieved at the Family and Genus levels. A unique genus and family could be delineated per order by phylopeptidomics. Finally, the Sphingomonas (Sphingomonadaceae), Caulobacter (Caulobacteraceae), and Acidovorax (Comamonadaceae) genera were validated with 340, 33, and 16 unique taxon-specific peptides, respectively. Their respective quantities may be roughly assessed from their TSM signal: 80% (898), 11% (122) and 9% (96). The proportion of each genus would be significantly different if based on the number of taxon-specific peptides: 87% (340), 8% (33), and 4% (16). Indeed, the number of taxon-specific specific peptides can vary significantly due to the phylogenetic relationships between the organisms present in the sample and the density of sequenced genomes per phylogenetic branch. Therefore, it is not a reliable parameter for quantifying the ratio of organisms as discussed previously (eviously .Sphingomonas sp. Leaf257 (TaxID 1736309), Sphingomonas parapaucimobilis (TaxID 28213), Sphingomonas melonis (TaxID 152682), Caulobacter vibrioides (TaxID 155892) and Acidovorax sp. CF316 (TaxID 1144317). As the number of bacteria whose genome has been sequenced and whose species taxonomy is well established in these important groups of environmental bacteria is still rather low , we cannot ascertain at the present stage the accuracy of taxa identified at the species taxonomical rank. For example, either the presence of a strain with intermediate genome characteristics between Sphingomonas sp. Leaf257, S. parapaucimobilis, and S. melonis, or the mixture of three closely related species could explain the Sphingomonadales signal.While the decrease in the number of specific peptides is rather sharp along the taxonomical ranks considered , the TSMs parameter is rather stable . Thus, no other main contributor to the biofilm, such as an unsequenced branch of life of bacteria, should be considered to explain the proteomic signal. The identification of organisms at a lower taxonomic rank than the genus rank would require a larger experimental dataset, thus more starting material, and a more comprehensive database for the three genera identified. Because the biofilm may comprise several closely related species whose genome have not yet been sequenced and reported in the NCBInr database, the identification at the species level in this study is only speculative. The most closely related major species identified by their specific peptides were: Sphingomonas, Caulobacter, and Acidovorax, respectively. Regarding the TWW biofilm, a large quantity of MS/MS spectra was recorded. In this case, a rather low number of TSMs was assigned to the superkingdom Bacteria (676) compared to the BNW biofilm (1168). Anyway, the three genera present in this sample were unambiguously identified by phylopeptidomics: Sphingomonas, Caulobacter and Acidovorax, certified by 214, 18 and 8 taxon-specific peptides, respectively. This shows the relative homogeneity of the biofilms in the pool, with the exception of the BWW biofilm as presented below.Alphaproteobacteria), with a large number of taxon-specific peptides (703). The Betaproteobacteria class counts only three specific peptides, which is below the validation threshold. Two orders, Sphingomonadales and Caulobacterales, contributed to this signal. The genera identified were: Sphingomonas and Caulobacter, with 497 and 7 taxon-specific peptides, respectively. Therefore, the BWW biofilm stands out from the others as no evidence of Acidovorax could be detected in this biofilm although a larger amount of material was available. At the species taxonomical rank, the most-closely related species identified by their specific peptides in all the sampled biofilms were the same as those identified for the BNW biofilm, with the exception of BWW, which does not comprise any detectable Acidovorax signal. Correlating these results with the radiation doses received by the different biofilms, irradiation of the north wall five months before sampling had no further effect on the microbial community in the BNW biofilm. In contrast, irradiation of the west wall in the three weeks prior to sampling could explain Acidovorax disappearance from the BWW biofilm community.The latest biofilm sample, BWW, provided the largest MS/MS spectra data set, i.e., corresponding to the most material-rich sample. In this case, only one class was validated  but explaining a Burkholderiales MS/MS-certified signal (nine taxon-specific peptides). The BWW biofilm signal differed from the others due to the absence of Betaproteobacteria phylopeptidomics signal. Some slight discrepancies in terms of relative ratio of each genus were noted between the different taxonomic ranks (ic ranks . HoweverSphingomonas (490), while proteins from Acidovorax and Caulobacter were less detected . The remaining 60 proteins were attributed to contaminants such as keratins. The numbers of protein groups identified in the four biofilms were relatively different: 340 (BNW), 173 (TNW), 388 (BWW), and 217 (TWW), respectively. In total, 71 protein groups were common to all four samples. Within this subset, the most common bacterial proteins were the elongation factors Tu and G involved in translation, the molecular chaperone GroEL, the subunit beta of DNA-directed RNA polymerase, and both the alpha and beta subunits of ATP synthase. This suggests that the cells from the three genera were metabolically active.Although the signal was relatively low, metaproteomics interpretation could be achieved using a reduced pan-proteomics database encompassing the protein sequences of all genome-wide sequenced strains belonging to the three genera detected in the samples and usual contaminants in proteomics experiments. This pan-proteomic database included a total of 498,167 protein sequences from 4,032 different annotated genomes. By merging the MS/MS datasets from the four biofilms, a total of 3,165 peptides were identified and 695 proteins were validated , 2. BaseSphingomonas were the flagellar motor proteins MotB and MotA, several TonB-dependent transporters (x8), several catalases (x8), several ribosomal proteins , the cell division protein FtsZ, and several enzymes of the central metabolism . Several TonB-dependent transporters (x3) and the two flagellar motor proteins MotB and MotA were also identified for Caulobacter, as well as two subunits of the nitrate reductase A . Specific proteins were identified in Acidovorax such as porins (x4), glutamate dehydrogenases (x2), outer membrane protein W, outer membrane protein/peptidoglycan-associated lipoprotein, alkyl hydroperoxide reductase, and glutamine synthetase. Interestingly, several proteins potentially linked to DNA repair were observed in Sphingomonas  and Caulobacter (RmB DNA/RNA helicase). Furthermore, two DNA-binding response regulators (OmpR and CitB–like transcription regulator proteins) have been identified in Sphingomonas. In several microorganisms, the SOS regulon encodes proteins involved in the coordinated response to DNA damage , which can theoretically be obtained by sequencing the equivalent of a nucleic acid sequence totaling 42 trillions of nucleotide positions. However, due to the history of Life starting from a common ancestor, the molecular relationships between organisms and the constraints of protein folding for stable structure and function, the sequences possibilities are more limited than this theoretical estimation. Anyway, the taxonomic identification based on the biofilm dataset collected in the present study is highly reliable because it is based on a large number of taxon-specific peptides. As previously demonstrated, metaproteomics and metagenomics deliver an identical taxonomic picture of complex samples because of the high density of molecular information analyzed  for facing the extreme environmental conditions encountered in Antarctica, i.e., hyper aridity, UV radiation and cold temperatures  was proven to be tolerant to 200 μM of uranium at pH 7 (Caulobacter has been identified in spent nuclear fuel storage pools in Sellafield (UK) (Caulobacter bacteria are well known for their distinctive ability to live in low-nutrient environments (The genus nd soils  and has nd soils . Its pre at pH 7 . The geneld (UK) . Caulobaronments . DetectiAcidovorax genus have been isolated from lawn soil, water, commensal flora, activated sludge plant water and have also been described as phytopathogen  while on the other hand, Caulobacter metabolism is oriented to specific metabolic process typical of stationary phase .The functional analysis performed with gene ontology terms reveals striking differences between the two main microorganisms. On the one hand, In conclusion, we have shown in this work that the microbial composition of biofilms can be directly analyzed using metaproteomics from as little as 0.02 mg of biological material. An original method for remote sampling of biofilms is presented, which can be employed to investigate other hostile environments. An innovative methodology for the processing of the MS/MS spectra data obtained after protein extraction has been developed for the proteotyping of microorganisms present in biofilms. The strategy developed here can be used to identify microorganisms present in any kind of samples and to decipher their respective contributions to the biomass. A last asset of proteomic analyses is that they provide direct access to the functioning of microorganisms present in microbial communities.doi: 10.6019/PXD039499 and are publicly available (http://www.ebi.ac.uk/pride/archive/projects/PXD039499).The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD039499 and OP, PP, CR, and JA: study conception and design and draft manuscript preparation. PP, GS, CR, and JA: data collection. All authors contributed to analysis and interpretation of results, reviewed the results, and approved the final version of the manuscript.


After COVID-19, there was an outbreak of a new infectious disease caused by monkeypox virus. So far, no specific drug has been found to treat it. Xuanbai Chengqi decoction (XBCQD) has shown effects against a variety of viruses in China.We searched for the active compounds and potential targets for XBCQD from multiple open databases and literature. Monkeypox related targets were searched out from the OMIM and GeneCards databases. After determining the assumed targets of XBCQD for monkeypox treatment, we built the PPI network and used R for GO enrichment and KEGG pathway analysis. The interactions between the active compounds and the hub targets were investigated by molecular docking and molecular dynamics (MD) simulations.In total, 5 active compounds and 10 hub targets of XBCQD were screened out. GO enrichment and KEGG analysis demonstrated that XBCQD plays a therapeutic role in monkeypox mainly by regulating signaling pathways related to viral infection and inflammatory response. The main active compound estrone binding to target AR was confirmed to be the best therapy choice for monkeypox.This study systematically explored the interactions between the bioactive compounds of XBCQD and the monkeypox-specific XBCQD targets using network pharmacological methods, bioinformatics analyses and molecular simulations, suggesting that XBCQD could have a beneficial therapeutic effect on monkeypox by reducing the inflammatory damage and viral replication via multiple pathways. The use of XBCQD on monkeypox disease was confirmed to be best worked through the estrone-target AR interaction. Our work could provide evidence and guidance for further research on the treatment of monkeypox disease. In JuneMonkeypox disease is a rare zoonotic disease caused by the monkeypox virus . It is eThe monkeypox virus is a double-stranded DNA virus that is related to the variola virus . The cliXuanbai Chengqi decoction (XBCQD) is a TCM consisting of four Chinese herbs, including mineral-based gypsum, herbal rhubarb, bitter almond, and trichosanthes . It was In fact, XBCQD is a representative Chinese medicine prescription in the Differentiation of Febrile Disease written by Wu Jutong in Qing dynasty. It has been widely used in China for the treatment of lung injury, pulmonary fibrosis, chronic obstructive pulmonary and other common respiratory diseases. It can effectively reduce phlegm, heat, cough, wheezing, chest tightness, and has less adverse reactions . With thin vivo binding patterns of ligand-receptor  of the targets.2.3.The targets of monkeypox disease were searched by using OMIM2.4.Using the Venny online database,2.5.The String2.6.Homo sapiens” in “List.” The target, the minimum overlap, the p-value and the minimum concentration were set to “human,” 3, 0.05 and 1.5, respectively. Then we ran the gene ontology function and KEGG pathway on the target genes. We screened and preserved the results of the most important BP, CC, MF, and KEGG pathways. Finally, the corresponding bubble diagram was derived.GO function and KEGG path enrichment were analyzed using Bioconductor2.7.To reveal the binding patterns between the active compounds and the targets, AutoDock Vina software, Discovery Studio 4.5 Client and PyMOL software were used. The 3D structures of the central targets (receptors) were obtained from the RCSB PDB2.8.The long-range electrostatic interactions were calculated using the particle mesh Ewald (PME) method. The target was placed in the center of simulation box filled with water TIP3P molecules with distance of 1.2 nm from the box boundary. There were 33 Na+ ions introduced in the water box to neutralize the charge of the whole system. The system was firstly balanced with energy minimization process which ran up to 100,000 steps using the steepest descent algorithm. Then the system was equilibrated to 310 K with v-rescale  method and backbone restrained. Subsequently, the system was further equilibrated at constant pressure (1 bar) and constant temperature (310 K). Finally, 200 ns MD test was conducted after all the restraints released. The simulation results were analyzed by Gromacs 5.1.2 built-in tools and our in-house scripts.3.3.1.In total, 285 compounds were screened out by the TCMSP database, among which 16, 19 and 11 compounds were from herbal rhubarb, bitter almond, and trichosanthes, respectively. And 43 active compounds met the OB ≥ 30% and DL ≥ 0.18 standards, which were selected for further analysis after removal of the duplicates. The TCMSP database was used to obtain the corresponding targets. All targets were then entered into the Uniprot database and normalized by removing the repeated ones. Finally, 116 potential targets were obtained.3.2.The GeneCards database  and the 3.3.According to the PPI network analysis , the 36 The ranking of the nodes of the most important active compounds in XBCQD was summarized in 3.4.p value of the BP item.The 36 XBCQD-monkey pox virus intersection targets were imported into the Metascape platform. GO functional enrichment analysis was performed on the targets of the active ingredients in the treatment of monkey pox virus from the levels of BP, CC, and MF . The topp value in p value of the path.It can be found that BP was related to rhythmic process, positive regulation of cell cycle, response to drug, epithelial cell proliferation, DNA-templated transcription, initiation, response to steroid hormone, regulation of epithelial cell proliferation, negative regulation of protein phosphorylation, regulation of epithelial cell proliferation, and transcription initiation from RNA polymerase II promoter. There were 155 items in the CC analysis , and thep values from the KEGG enrichment analysis and related targets were used to construct the KEGG key pathway network , Stigmasterol (MOL003035), Gamma-Aminobutyric Acid (MOL000388), Phytol (MOL001442) and estrone (MOL008204). The protein structures of hub targets were acquired online from RCSB PDB, including AR (PDB ID: 2QPY), CASP3 (PDB ID: 6BDV), CCNB1 (PDB ID: 2B9R), CDKN1A (PDB ID: 3TS8), ESR1 (PDB ID: 1A52), IL1B (PDB ID: 4DEP), JUN (PDB ID: 1FOS), MYC (PDB ID: 7C36), TP53 (PDB ID: 4MZI) and TNF (PDB ID: 1TNF). The values of affinity energy were obtained by docking analyses . Notably3.6.The two best dockings between the compounds and the targets (estrone-AR and estrone-ESR1) were selected to perform the MD simulations. After 200 ns of MD simulations, the dynamic evolutions of estrone-AR and estrone-ESR1 complexes were studied. The conformations of estrone-AR and estrone-ESR1 complexes as well as their contact residues were shown in As shown in From 4.TCM has accumulated a long history of clinical experience and efficacious prescriptions in preventing and treating diseases . It has The PPI analysis of the 36 targets showed that the top 10 hub targets, including ESR1, TP53, CASP3, JUN, TNF, MYC, CDKN1A, 1L1B, CCNB1, and AR may be the key targets of the treatment of monkeypox virus. The top 5 beta-sitosterol, Stigmasterol, Gamma-Aminobutyric Acid, Phytol, estrone and the top 10 targets were selected in this study. These bioactive compounds were found to effectively bind to the 10 targets according to the results of molecular docking. We further performed GO enrichment analysis on the 36 selected targets to better understand the multiple effects of XBCQD against monkeypox virus from a systematic perspective. The top 10 GO functional categories indicated that XBCQD may exert its effects through the involvement in the BP, MF, and CC. Among these targets, TP53 and ESR1 ranked highest. It was reported that TP53 could be one of the key enzymes in prostaglandin biosynthesis, which had an association with inflammation and mitosis . In addivia these pathways.The concentration of GO terms in hub targets showed that XBCQD treatment of monkeypox was mainly involved the rhythmic process, positive regulation of cell cycle, response to drug, epithelial cell proliferation and DNA-templated transcription, etc. KEGG pathway enrichment analysis of these targets indicated that they were involved in Hepatitis B, AGE-RAGE signal pathway of diabetes complications, Breast cancer, Proteoglycans in cancer, Human cytomegalovirus infection, etc. The AGE-RAGE signaling pathway was reported to have a regulatory role in diabetes . Some stThe results of network pharmacology were validated by molecular docking of the top 10 targets and 5 active compounds. The two complexes with lowest binding energy scores were found to be estrone-AR and estrone-ESR1. Furthermore, MD simulations showed that the average value of interaction energy of estrone-AR was −186 kcal/mol , while was lower than the value of −157 kcal/mol  of the estrone-ESR1 complex. Therefore, the interaction of estrone-AR complex was more stable comparing to estrone-ESR1 complex.via multiple pathways. In addition, the use of XBCQD on monkeypox disease was confirmed to be best worked through the estrone-target AR interaction. By using a series of computational approaches, our study established the drug screening of XBCQD on monkeypox disease for the first time, hopefully could provide some guidance for future drug development.In summary, this study systematically explored the interactions between the bioactive compounds of XBCQD and the monkeypox-specific XBCQD targets using network pharmacological methods, bioinformatics analyses and molecular simulations, suggesting that XBCQD could have a beneficial therapeutic effect on monkeypox by reducing the inflammatory damage and viral replication 5.By combining network pharmacology, molecular docking and MD simulation, the molecular mechanism of XBCQD in the treatment of monkeypox virus was systematically investigated. According to our results, the top 5 compounds  and the top 10 targets  were identified. We also found that XBCQD in the treatment of monkeypox virus could be mainly related to Hepatitis B, AGE-RAGE signal pathway of diabetes complications, Breast cancer, Proteoglycans in cancer, Human cytomegalovirus infection, MAPK signaling pathway, Estrogen signaling pathway, Epstein–Barr virus infection, Human immunodeficiency virus 1 infection and Chemical carcinogenesis-receptor activation. Moelcular docking showed that estrone-AR and estrone-ESR1 exhibited stronger binding affinities than the other moieties. And the estrone-AR possessed higher structural and interaction stabilities than the estrone-ESR1 with the extension of molecular simulation time. Our study provides a comprehensive explanation of the multi-component, multi-target and multi-pathway intervention mechanism of XBCQD in the treatment of monkeypox, which is expected to provide a basis and new insights for further pharmacological research.The original contributions presented in the study are included in the article/YS designed the simulations. YJ and CS performed the simulations and analyzed the data. YS, YJ, and CS wrote the manuscript. All authors contributed to the article and approved the submitted version.This work is supported by the National Natural Science Foundation of China (Ref: 12102113) and the Major program of the National Natural Science Foundation of China (T2293720/T2293722).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Background: Trigeminal neuralgia (TN) is defined as a spontaneous painful sensation in the trigeminal nerve territory. The pain intensity of TN is classified into different grades of suffering that affect a patient’s quality of life. Percutaneous balloon compression of the ganglion is a neurosurgical option that is easy, reproducible, and can reduce the morbidity of TN.Methods: We retrospectively analyzed all patients treated with trigeminal nerve percutaneous balloon compression at the Clinical Hospital of the University of Chile between January 2012 and May 2022. Data collected from electronic records included demographic information, medical and surgical history, type of anesthesia and drugs used during surgery, balloon inflation time, surgery time, operative room time, intraoperative events, postoperative complications, duration of hospitalization, and duration of follow-up. Results: We identified 63 patients who met our inclusion criteria. The median patient age was 62 years (interquartile range [IQR] 57-69 years). Sixty-five percent of the patients were female. The simultaneous involvement of the second and third branches of the trigeminal nerve was the most frequent symptom. Before surgery, the patients experienced an average of 6.6 years of pain (IQR 2-10 years). Right neuralgia was the most frequent laterality type (69%). Forty percent of the patients had a previous surgical procedure for neuralgia, with treatment failure being the most frequent surgical indication (63%). According to the procedure, the mean balloon insufflation volume was 0.89±0.12 mL with a median compression time of 2.5 min (IQR 2.1-4.0 min). No hemorrhagic complications were observed. Furthermore, during follow-up, there were no surgical complications among any of the patients; however, 6.4% of patients required a second intervention. The pain-free period was two years in 60% of patients and five years in 23% of patients.Conclusions: TN is a painful condition. Although there are multiple surgical approaches, we believe that percutaneous balloon compression is an excellent alternative treatment option that offers high effectiveness, low morbidity, and low hospital stay.  Trigeminal neuralgia (TN) is defined as a spontaneous painful sensation in the trigeminal nerve territory. Pain intensity involves different grades of suffering that affect the quality of life. The main criteria for diagnosis are brief, intense, and sudden paroxysms of pain triggered by innocuous stimuli such as talking, chewing, or light touch of the overlying skin, and is restricted to one or more branches of the trigeminal nerves . AlthougTreatment options include medications and surgery. The initial management of choice for TN is medical pharmacotherapy. Patients who do not tolerate or fail medical management may benefit from surgery ,4. The oIn general, all techniques have an initial response of more than 80% at five years, and MVD has a superior response compared to the other techniques. Significant risks associated with MVD occur at rates of less than 2% and include stroke, meningitis, and death .Percutaneous balloon compression (PBC) of the Gasserian ganglion (GG) is an alternative neurosurgical treatment option. The technique involves needle insertion through the foramen ovale under the GG. With the needle tip placed in Meckel's cave, a Fogarty catheter is inserted, and balloon insufflation with contrast material is performed under direct radioscopic vision. The balloon is loaded with up to 1 mL of the solution, compressing the ganglion for 1.3-3 minutes (Mullan technique). Compression of trigeminal fibers allows for immediate pain relief in 80-90%. Advantages of this technique include high effectiveness, a low hospital stay, and minimal morbidity and mortality .In 1991, we started treating TN using PBC of the GG. Here, we present the results of a 10-year retrospective review (2012-2022) at the Clinical Hospital of the University of Chile.This article was presented as a poster at our Chilean neurosurgical meeting in September 2022.Study design and settingA retrospective study was conducted using digitized records of patients who underwent PBC of the trigeminal nerve at the Clinical Hospital of the University of Chile between January 2012 and May 2022. The study received approval from the ethics committee (Ref: OAIC:33/2022). All personal information was handled confidentially.A search was conducted within the electronic clinical records of all surgeries identified using the terms "Percutaneous balloon compression" and "Neurolysis with microsurgical technique." Patients who underwent alternative procedures, including peripheral neurectomies, percutaneous glycerol rhizotomy, percutaneous radiofrequency rhizotomy, radiosurgery, and MVD, were excluded from the analysis.Data collected from electronic records included demographic information, medical and surgical history, type of anesthesia and drugs used during surgery, balloon inflation time, surgery time, operative room time, intraoperative events, postoperative complications, duration of hospitalization, and follow-up time of patients Table .Quantitative variables and statistical methodsCategorical variables are summarized as relative frequencies. Continuous variables were expressed as means (and standard deviations) or medians (and interquartile ranges [IQR]). We found 63 patients treated with PBC within the 10-year lookback period  was the most frequent symptom, followed by the independent involvement of each of the territories. Patients were treated after 6.6 years of pain (IQR 2-10 years), with right neuralgia as the most common laterality (69%). Medical treatment failure was the most frequent surgical indication (63%), and in the 10-year study period, the recurrence rate was 25%.All procedures were performed under general anesthesia. Propofol/remifentanil site-based target control infusion was the most frequent technical procedure for the administration and delivery of anesthesia.All surgeons performed the same procedure Table , 2. DuriAfter surgery, only one patient presented with pain and 16 (25%) had hypoesthesia or paresthesia of the V2-V3 territory. The mean postoperative stay was one day. No hemorrhagic complications were observed in our case series.Based on our records, during a median follow-up period of four months, no patient experienced any surgical complications; however, 10 patients presented with pain recurrence that required a second PBC procedure after 2.7 years.Finally, a phone interview with the entire cohort was performed. Of the 42 patients who had their first surgery at our center, 29 responded to the telephone evaluation. Of these, 18 patients showed evolution to mild facial hypoesthesia. Additionally, three patients required a new surgery. Of these, one patient was pain-free for three months, one patient for one year, and one patient for two years before requiring a second surgery.TN is a benign disease that does not modify a patient's life expectancy but has a meaningful impact on daily living activities. In this study, we reported our 10-year single-center experience with PBC of the GG for the treatment of TN.Although TN is a rare condition, affecting 4-13 people per 100,000 annually, it is the most common facial neuralgia. It is more common in women 2:1), with an incidence that increases with age and is more frequent in the fifth or sixth decade of life [:1, with Neurological examination and neuroimaging should exclude secondary causes, such as vascular malformations, multiple sclerosis, or expansive processes (secondary TN). Angio-magnetic resonance allows anatomical structure visualization of the trigeminal nerve and its relationships and is especially useful for ruling out neurovascular compression that may lead to posterior fossa surgical exploration, particularly in younger patients ,6-8.The pathophysiological origin of classic and secondary TN is nerve damage at its entry into the pons, and the trigeminal nerve  loses its Schwann cell myelin sheath, which is replaced by central myelin generated by oligodendrocytes. This transition zone is vulnerable to damage, particularly demyelination. Demyelination at these sites has been demonstrated by neurophysiological, neuroimaging, and histological studies . When thPharmacological and surgical management are used as TN treatment modalities Table . MedicalSurgical intervention for TN patients is necessary whenever adverse effects or symptoms are not abated with medical treatment. These options include MVD, PBC, percutaneous radiofrequency rhizotomy, percutaneous glycerol rhizotomy, and stereotactic radiosurgery. The most used techniques are MVD and PBC, which are strong, viable alternative options due to the low morbidity associated with these techniques. In general, all techniques have an initial response of more than 80% at five years .MVD is a suboccipital craniotomy. It is the most invasive technique but has a superior response at a 10-year follow-up compared with the other percutaneous techniques. Significant risks associated with MVD occur at rates of less than 2% and include stroke, meningitis, and death, but in experienced hands, the complication rate is relatively low .Radiofrequency rhizotomy provides a high initial success rate of 90%, though recurrence occurs in 25% of cases, with a long-term response rate of 57.7% at five years. PBC provides immediate pain relief in 80-90% of patients, with a long-term response rate of 69-80% at five years ,2,16-18.An interesting finding was that patients underwent surgery with 6.6 years of symptoms, probably due to low suspicion and low referral to specialists (neurologist or neurosurgeon). Furthermore, in the 10 years reviewed, we recorded a recurrence rate of 25%, which is compatible with the literature. These patients underwent a second surgery in an average of 2.7 years. PBC of the GG damages the thick myelinated fibers (which trigger pain by ephaptic-type contact with nociceptive fibers), usually leaving the thin and unmyelinated myelinated fibers unharmed ,9,10. FiDuring compression, pear-shaped balloon insufflation indicates adequate balloon pressure in the cavity. An hourglass image denotes balloon protrusion into the prepontine cistern. In such cases, the catheter is deflated, removed a few millimeters, and again insufflated. Exaggerated balloon insufflation compresses the upper inner wall of the cavum, with cranial nerve VI involvement. In our series, we observed a first-attempt, pear-shaped image in more than 90% of the patients.Although percutaneous techniques are "less invasive," they also carry a risk of loss of sensitivity in the trigeminal territory, dysesthesia, loss of corneal reflex, painful anesthesia, meningitis, subdural hematoma, and a very low risk of mortality ,21-24.If a patient's pain is refractory to PBC performed using the proper technique, it is necessary for the practitioner to review the patient’s diagnosis. Patients with long-standing neuralgia and treatment-resistant trigeminal naturopathy can develop atypical facial pain.TN is a severely painful and disabling condition. Our 10-year experience with GG PBC highlights its efficacy after medical treatment failure. We observed high effectiveness and low morbidity, as evidenced by the absence of surgical complications and minimal symptomatic recurrence. While MVD remains the gold standard, our analysis suggests that PBC presents a compelling and easily implementable alternative for neurosurgical teams.As a team, it seems interesting for future studies to protocolize the postoperative management of patients based on pain scales and local protocols in order to offer different surgical options depending on each patient. Even so, sufficient data have not yet been obtained and further studies are needed.


E2F8, FOXM1, and MYBL2 has broad applicability, with an AUC value of up to 0.877 at one year. As a high-risk clinical factor, its abnormal disorder may lead to upregulation of the activity of pathways related to cell proliferation. This model can also be used to predict the adverse effects of immunotherapy in patients with TNBC. Molecular docking was used to screen three drugs that target TFs: Trichostatin A (TSA), Doxorubicin (DOX), and Calcitriol. In vitro and in vivo experiments showed that TSA + DOX was able to effectively reduce DOX dosage, and TSA + DOX + Calcitriol may be able to effectively reduce the toxic side effects of DOX on the heart. In conclusion, the machine learning model based on three TFs provides new biomarkers for clinical and prognostic diagnosis of TNBC, and the combination targeted drug strategy offers a novel research perspective for TNBC treatment.Transcription factors (TFs) have been shown to play a key role in the occurrence and development of tumors, including triple-negative breast cancer (TNBC), with a worse prognosis. Machine learning is widely used for establishing prediction models and screening key tumor drivers. Current studies lack TF integration in TNBC, so targeted research on TF prognostic models and targeted drugs is beneficial to improve clinical translational application. The purpose of this study was to use the Least Absolute Shrinkage and Selection Operator to build a prognostic TFs model after cohort normalization based on housekeeping gene expression levels. Potential targeted drugs were then screened on the basis of molecular docking, and a multi-drug combination strategy was used for both in vivo and in vitro experimental studies. The machine learning model of TFs built by  Triple-negative breast cancer (TNBC) is a highly invasive and heterogeneous subtype of breast cancer that accounts for approximately 10–20% of breast cancer . Breast ACTB, GAPDH, TFRC, TUBB) to normalize the expression levels of nine Hub TFs genes across the training and validation groups. The Least Absolute Shrinkage and Selection Operator (LASSO) regression algorithm was used to build the Hub TF feature score system (HTFSS), consisting of E2F8, FOXM1, and MYBL2 genes. The sensitivity of HTFSS for prediction of prognosis was high, with 1-year Area Under Curve (AUC) values as high as 0.877 and 0.987 in the training and validation cohorts, respectively. Our assessment of multiple clinical features demonstrated that HTFSS can be used as an independent clinical risk factor for predicting worse patient prognosis and poor outcomes with immunotherapies. To extend the clinical application of HTFSS, a Nomogram was also constructed, and an online version of the interactive webpage was deployed to aid clinicians in their diagnosis. Based on scRNA-seq cohorts, we investigated the relationship between HTFSS and the tumor immune microenvironment (TIME). We found that E2F8 and FOXM1 were significantly associated with MKI67+ progenitor cells, while MYBL2 was significantly associated with dendritic cells of the lymphoid lineage. Multiple TF target gene databases were used to explore the mechanism of regulation of the downstream target genes, and changes in pathway activity caused by imbalances in the three TFs were explored. Interestingly, when the three TFs were dysregulated, cell proliferation-related pathways, such as the PI3K-AKT and FOXO signaling pathways, were upregulated. Next, by combining the Comparative Toxicogenomics Database (CTD) and molecular docking, three targeted drugs (Doxorubicin (DOX), Trichostatin A (TSA), and Calcitriol) with strong binding activity to the expression proteins of the Hub TF gene were tested. We next tested the efficacy of the combination treatment in the murine model and at the cellular level. At the cellular level, RT-qPCR results showed that all three TFs were significantly upregulated in TNBC cell lines, and both DOX and TSA were able to effectively reduce their abnormal expression. DOX and TSA can also inhibit tumor cell progression by inhibiting the expression of all three TFs and regulating the PI3K-AKT and FOXO signaling pathways. In vivo and in vitro experimental results demonstrated that the combination of TSA + DOX was capable of effectively reducing the DOX dosage, which was beneficial to patients because it has been reported that DOX has multiple toxic side effects. TSA + DOX + Calcitriol can be used in combination to effectively reduce the toxic side effects of DOX on the heart, which is further optimization of the DOX, reducing the toxic side effects of the drug based on enhancing the antitumor effect. In conclusion, we believe that the clinical application of HTFSS provides a more sensitive and widely applicable biomarker of TFs integrated for TNBC, and the strategy of combined use of targeted drugs has the potential to optimize the antitumor effect of a single drug to some extent.In this study, bulk RNA-seq and single-cell RNA-seq (scRNA-seq) technologies were combined to conduct an exhaustive study of TFs in the TNBC cohort, and the specific research workflow is shown in First, 3396 differentially expressed genes (DEGs) were obtained from TCGA-BRCA, of which 1975 were upregulated and 1394 were downregulated A. WGCNA MYBL2Exp − 0.010 × FOXM1Exp − 0.295 × E2F8Exp. Next, the predictive value of the HTFSS was evaluated by the time-dependent Receiver Operating Characteristic (ROC) curve. The results demonstrated that the AUC value for the patient’s first-year survival rate was as high as 0.877, and the AUC values for survival at 3, 5, and 7 years were 0.699, 0.587, and 0.663, respectively  based on the underlying formula of the Nomogram  results revealed that low HTFSS levels had a higher immune score A. HTFSS In this study, we performed low-resolution dimensionality reduction and cluster analysis on scRNA-seq cohorts of 10 patients with TNBC. We obtained 12 major cell clusters A, consisE2F8  and TSA (0.6 μM), the expression of both FOXM1 and MYBL2 was downregulated, and the expression of E2F8 was upregulated in the presence of DOX treatment  and TSA (0.6 μM) for 48 h in the presence of MDA-MB-231. Based on Western blotting detection results, DOX and TSA were shown to significantly decrease mTOR and PI3K protein expression levels and the levels of phosphorylation of P70, S6, and 4EBP1 within the PI3K-AKT signaling pathway  and the type 1 trichorhinophalangeal syndrome (TRPS-1), as markers of breast cancer diagnosis by IHC, have been included in TNBC [BACH1 regulates cancer cell invasion genes, aiding in the migration and invasion of breast cancer cells, whereas acetyl tricyclic bis (cyanoketone) can produce novel activity and destroy BACH1 expression [TNBC is among the most prevalent female cancers worldwide, with the hallmarks of high deterioration, high invasiveness, high heterogeneity, and a poor prognosis. As the largest branch of the artificial intelligence field, machine learning has invaluable opportunities in the diagnosis and prediction of TNBC . As crit in TNBC  diagnostpression . The preACTB, GAPDH, TFRC, and TUBB) and three TFs , which had an excellent ability to predict prognosis. In the training and validation cohorts, the AUC values at 1 year were 0.877 and 0.987, respectively. Similarly, the results of the Cox regression analysis showed that the HR for the HTFSS was much greater than one, which may serve as an independent clinical risk factor. Applying HTFSS to different clinical features of TCGA, HTFSS was found to be significantly correlated with the N stage of the tumor. This means that the higher the HTFSS, the worse the patient’s lymph node involvement. Studies of immune infiltration and immunotherapy cohorts have also shown that HTFSS positively correlates with poor immunotherapy outcomes. This means that the higher the HTFSS, the worse the patient’s immunotherapeutic effect. We conclude that the HTFSS developed by us based on TFs has a great deal of clinical prognostic value, which may not only predict patient prognosis and lymph node involvement but also the effectiveness of immunotherapies. More importantly, we have developed a Nomogram-based online interactive Web page that can be applied to clinical diagnostics by detecting the expression levels of only four housekeeping genes and three genes encoding TFs, greatly enhancing the translation of clinical research. Subsequent studies demonstrated that HTFSS affected the occurrence and development of TNBC and TIME through the regulation of pathways related to MKI67+ progenitor cell and PDCs proliferation. To suppress the prognostic issues caused by the imbalance in HTFSS, we developed a combination strategy and screened three targeted drugs, DOX, TSA, and Calcitriol, by molecular docking and studied their antitumor effects. We found that DOX and TSA effectively inhibited E2F8, FOXM1, and MYBL2 overexpression and the abnormal expression of PI3K-AKT and FOXO signaling pathways. DOX + TSA combination therapy can effectively reduce DOX dosing. The combination of DOX + TSA + Calcitriol can effectively reduce cardiotoxic side effects caused by DOX while ensuring the antitumor effect, which undoubtedly led to the optimization of the clinical treatment plan for TNBC patients. In summary, studies on the prognostic mechanism and the combination of targeted drugs based on HTFSS provide a new perspective on the molecular mechanism and clinical treatment of HTFSS. However, there are also some ethical considerations when using HTFSS for clinical applications. Undeniably, the machine learning model has some limitations as well. Thus, HTFSS guidance for clinical diagnosis can only be used as a reference and cannot be relied upon for the final conclusion. Other clinical indicators must be considered to determine the specific drug response and prognosis of patients.The purpose of this study was to use machine learning to construct HTFSS highly correlated with TNBC, which was made up of the ratio of four housekeeping genes  were screened, and a new scheme for combined therapy was proposed. Calcitriol is a synthetic physiologically active Vitamin D analog that can induce cell cycle arrest in the G0/G1 phase, promote cell differentiation, and induce apoptosis, thereby inhibiting the proliferation of tumor cells. It is reported that Calcitriol can be used to treat advanced prostate cancer, hypercalcemia, and hyperphosphatemia ,28. DOX aterials ,35, whicaterials . In termLastly, this study has some limitations. The sample sizes of both Bulk RNA-seq and scRNA-seq cohort studies were small, leading us to ignore some information. Thus, in future joint analyses of multiple omics, we can consider using the strategy of multi-sample integration to broaden the search scope. Second, this study revealed that dysregulation of HTFSS can affect changes in the activity of four key pathways. The precise molecular mechanism remains to be explored, and further experimentation is required to verify these findings, which will be the focus of future work. Furthermore, the combination drug strategy remains in the preliminary exploratory stage, and further optimization is required in future research work, such as dose control of combination drugs and assembly of targeted drug delivery.We downloaded the TCGA-BRCA  from UCSp-value < 0.05. Using DEGs as the input of WGCNA [The patients with TCGA-BRCA were divided into tumor and normal groups for differential analysis by the DEseq2 package . A thresof WGCNA , the genof WGCNA  in R, whof WGCNA ,43, tookACTB, GAPDH, TFRC, and TUBB). After the HTFSS was constructed, the predictive performance of the scoring system was evaluated by the AUC value of the area under the time-dependent ROC curve.LASSO regression  is a macnormalized = APM − APMcancer_min × APMcancer_max − APMcancer_min; TIGS = APMnormalized × ln(TMB + 1).Univariate Cox and Multivariate Cox regression were used to study the relationship between HTFSS and clinical features. A Nomogram  was consThe scRNA-seq cohort was studied by the Seurat package (v4.0) in R. The mitochondrial gene content (percent. mt) was set to <15% to filter the single-cell matrix. The matrix was normalized; PCA dimensionality reduction was passed through the SNN algorithm (resolution = 0.4) to cluster cell clusters and display single-cell clusters with dimensionality reduction through the UMAP algorithm. Then, the FindAllMarkers function  was used to find the marker genes of all cell clusters. The cell clusters were annotated in a semi-supervised manner, that is, automated annotation through SingleR first, followed by CellMaker and PanglaoDB ,48 databWe downloaded the collection of targeted small molecule compounds from the CTD , the 3D In Bulk RNA-seq, the DEGs of the Hub TF gene were calculated using the scRNA-seq grouping method. Significant co-expressed genes were screened using Pearson correlation. Subsequently, the target genes of the Hub TF gene were downloaded from the databases of GTRD , Cistrom2. The medium was changed every 24 h, and experiments were performed with logarithmically growing cells. TSA (HY-15144) and Calcitriol (HY-10002) were purchased from MCE.Human triple-negative breast cancer cells MDA-MB-231 were cultured in DMEM medium  supplemented with 10% fetal bovine serum  and 1% penicillin/streptomycin and incubated at 37 °C and 5% COMDA-MB-231 cells were cultured to the logarithmic growth stage and treated with DOX, TSA, and Calcitriol at different concentrations in 48-well plates. Four groups were set up: the DOX group, the TSA group, the Calcitriol group, and the TSA and DOX group. After 48 h of administration, the cell medium was removed, and the cells were purified twice with 1x PBS. Complete medium and 10% detection reagent were added to the cells, and the cells were incubated at 37 °C for 30 min in the dark. The OD values of each well were detected with an enzyme marker at a 450 nm wavelength.RNA was extracted from cells. We collected the cell precipitates and added 1 mL of Trizol to lyse the cells for 5 min on ice. We added 200 μL of trichloromethane, mixed it by shaking, and placed it on ice. When the solution forms stratification, transfer it to a low-temperature centrifuge and centrifuge at 12,000 rpm for 10 min at 4 °C. The solution will form three RNA, DNA, and organic phase layers. Then, we transferred the upper RNA layer to a new 1.5 mL EP tube and added 0.8 times the volume of isopropyl alcohol. After inverted mixing, we stored the solution in the refrigerator at −20 °C for 30 min and then centrifuged again at a low temperature and 12,000 rpm for 10 min. The supernatant and purification of the RNA precipitation with 75% and 95% ethanol were discarded. After the remaining ethanol was absorbed and volatilized, we added 30–50 μL of DEPC water to dissolve the RNA precipitation. The quality and concentration of the extracted RNA were detected.cDNA synthesis by reverse transcription. The RNA was converted into cDNA using the Hifair III 1st Strand cDNA Synthesis SuperMix for qPCR (Yeasen Biotechnology (Shanghai) Co., Ltd., Shanghai, China) reverse transcription kit. The system for reverse transcription was configured according to the instructions provided by the kit. After the run, the resulting cDNA was used for subsequent RT-qPCR experiments.GAPDH was used as an internal reference gene. Data were calculated using the 2−∆∆Ct method. Primer sequences for the Hub TF genes are shown in RT-qPCR. Gene expression was analyzed using the FastStart Universal SYBR Green Master (Roche) test kit. The system was configured according to the instructions provided by the kit, with three replicate wells per sample. After sampling, the 96-well plates were put into the fluorescence quantitative PCR instrument for computer detection. MDA-MB-231 cells were treated with DOX (250 ng/mL) and TSA (0.6 μM) for 48 h. After treatment, the cells were collected and lysed in RIPA buffer to extract the protein. The protein concentration was determined using the BCA protein quantification method. Equal amounts of protein were subjected to electrophoresis with a 10% SDS-PAGE gel  and transferred to a PVDF membrane . The membrane was blocked with 5% skim milk or 2% BSA for 2 h and incubated overnight with the primary antibodies. These antibodies include GAPDH , β-Actin , Vinculin , PI3K , AKT , mTOR , P70 , S6 , 4EBP1 , p-P70 , p-S6 , p-4EBP1 , FoxO1 , FoxO3 , p-FoxO1 , p-FoxO3 . Then, the membranes were washed with Tris-buffered saline and Tween-20 for 10 min and incubated with secondary antibodies for 2 h at room temperature. The developer solution was prepared according to the horseradish peroxidase (HRP) instructions. The chemiluminescence signals were detected with enhanced chemiluminescence (ECL).The mice were purchased from Yunnan University, and adriamycin was purchased from Pu ‘er People’s Hospital. The BALB/C nude mice were kept in SPF houses, and their drinking water was changed every 2 days. Their food was added every 5 days, and their bedding was changed weekly. Twenty BALB/C nude mice were randomly divided into five groups: Control group, TSA group, DOX group, TSA + DOX group, and TSA + DOX + Calcitriol group. After one week of adaptive feeding, MDA-MB-231 tumor cells were mixed with matrix glue, and nude mice (1 × 106/cell) were injected in situ under the skin to construct the nude mouse orthotopic transplantation tumor model. After tumor formation, the nude mice were treated with intraperitoneal injections of DOX (2 mg/kg), TSA (1 mg/kg), and Calcitriol (0.5 μg/kg) once every three days, while the control group was treated with the same dose of normal saline. The changes in body weight and tumor size were recorded every three days during feeding. Twenty-one days after administration, the mice were killed by neck dislocation, and tumor, heart, and other tissues were collected for follow-up experiments.Clean female BALB/C nude 2O2 solution was added and incubated for 20 min at room temperature, away from light. Antigen repair was performed by boiling the sections in a 2% sodium citrate solution. To seal the sections, 5% goat serum was added. The primary antibody ki67  was added and incubated overnight at 4 °C. Next, the sections were incubated at 37 °C for 1 h with the HRP RABBIT/MOUSE  secondary antibody. DAB was used for the color reaction, followed by post-staining with hematoxylin for 1 min and conversion to blue using ammonia. After dehydration using an alcohol gradient, the sections were sealed with neutral gum, and the tumor tissue was viewed under a microscope.Fresh heart tissue was soaked in 4% paraformaldehyde for 24 h. After dehydration, transparency, and paraffin embedding, the paraffin block was sliced into sections of 4 μm thickness. The sections were dewaxed using xylene and an ethanol gradient, followed by inactivation. Next, a 3% HFresh heart tissue was soaked in 4% paraformaldehyde for 24 h. After dehydration, transparency, and paraffin embedding, the paraffin block was sliced into sections of 4 μm thickness. The sections were then dewaxed in xylene Ⅰ and xylene Ⅱ for 10 min each and rehydrated using a gradient of ethanol. Hematoxylin was used to stain the sections for 1 min, followed by hydrochloric acid differentiation and ammonia rehydration. Eosin was then used to stain the sections for 30 s, and after dehydration, the slides were sealed with neutral gum. The pathological changes in myocardial tissue were observed under a microscope.p-value < 0.05 indicated statistical significance.Statistical analysis was mainly performed by R language (v4.1.1) and GraphPad Prism (v8.0.2) in this study. The Wilcoxon test was used for statistical testing between different groups, and the Logrank test was used to test the significant difference in survival probability between samples. All of the experiments were repeated at least three times, with the data presented as means ± standard error of the mean (SEM) using one-way ANOVA to determine significant differences between multiple groups. E2F8, FOXM1, and MYBL2) in HTFSS were discussed from different angles. The three Hub TF genes are highly upregulated in TNBC and affect the proliferation of tumor cells through the PI3K-Akt signaling pathway, Wnt signaling pathway, FOXO signaling pathway, and mTOR signaling pathway, resulting in a poor prognosis. Targeted drug screening and virtual docking based on Hub TF genes provide a new idea for combined drug use. Cellular and mouse experiments have proved that TSA has anti-TNBC activity, and the combination of TSA and DOX can effectively reduce the dose of DOX. Although Calcitriol does not have anti-TNBC activity, the combination of TSA and DOX can significantly reduce the cardiac side effects caused by DOX. In conclusion, our study provides a new point of view on the prognostic diagnosis and targeted drug therapy of TNBC.In the present study, a machine learning model, HTFSS, which has outstanding prognostic and predictive value for immunotherapy efficacy, was established based on transcription factors. As an independent clinical high-risk factor, HTFSS can be used to predict lymph node involvement in patients with TNBC. A Shiny App based on HTFSS is helpful for clinical diagnosis. The pathway activity changes caused by the imbalance of three Hub TF genes (


In the intervention group, we found significant improvement in Pittsburgh sleep quality index of −2.49 points (95% CI −3.84 to −1.14) in comparison to the control group (p = 0.0003) and sleep efficiency measured with objective instruments . Conclusion: Our results found that physical exercise programs in older adults improve sleep quality and efficiency measured with objective instruments.Background: Given the beneficial effects of exercise in different populations and the close relationship between healthy ageing and sleep quality, our objective was to determine if physical exercise delivered through a structured program improves sleep quality in older adults. Methods: Embase, PubMed/MEDLINE, Web of Science, and Cochrane Register of Clinical Trials (CENTRAL) were searched to 15 January 2023. Studies that applied physical exercise programs in older adults were reviewed. Two independent reviewers analysed the studies, extracted the data, and assessed the quality of evidence. Results: Of the 2599 reports returned by the initial search, 13 articles reporting on 2612 patients were included in the data synthesis. The articles used interventions based on yoga ( The number of older people is growing worldwide. Currently, about 10% of the global population is over 65 years old . The chaSufficient lifetime physical activity, healthy food habits, and significant sleep quality are major contributors to healthy ageing ,7. On thSleep disorders, such as altered sleep duration and increased sleep fragmentation, are common among older adults . The mosPoor sleep quality in older adults is expressed, among other ways, as insomnia, a sleep–wake disorder characterised by difficulty initiating or maintaining sleep or both, and older adults are more likely to be prescribed medication for insomnia treatment . MedicatExercise is a widely accepted approach adopted to improve physical function, cardiovascular health, and mental health, and it may also be beneficial for sleep ,22,23,24Given the beneficial effects of exercise in different populations and the close relationship between healthy ageing and sleep quality, our objective was to determine if physical exercise delivered through a structured program improves sleep quality in older adults.A systematic review was reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines . The revRandomised controlled trials (RCTs) that included older adults (>65 yr)  performiThe Embase, PubMed/MEDLINE, Web of Science, and Cochrane Register of Clinical Trials (CENTRAL) were reviewed on 23 October 2022 and updated on 15 January 2023. Manual searches with the following terms were conducted: ((elderly) OR (older people) OR (older adults) OR (ageing)) AND ((Exercise) OR  OR  OR (resistance training) OR (aerobic) OR (gym) OR (gymnastic) OR (strength) OR ) AND  OR (sleepiness) OR (apnea-hypopnea index)) . No language or publication restrictions were imposed.The terms selected were combined using Boolean logical operators . Additionally, a manual search of the references included in the selected articles was conducted. All references were analysed using Rayyan web software .The selected articles were reviewed independently by investigators with experience in meta-analysis and training in the literature review. First, the titles and abstracts of all identified studies were reviewed in pairs by four investigators (LSN-MOY-OM-RTC). Studies deemed irrelevant based on the title and abstract were excluded. Any possible disagreements were resolved by a third reviewer (MSR). Second, the first step’s full-text versions of the articles selected were reviewed and rechecked against the eligibility criteria (LSN-RTC). Again, any potential disagreements were resolved by a third reviewer (MSR). There were no disagreements between the investigators. Finally, study authors were contacted as required to determine study eligibility.Two authors (OM-RTC) extracted the data using a standardised protocol and reporting forms. The following information was extracted from each included study: design, population characteristics, and exercise program characteristics . If some relevant data were not in the article, the author was contacted to request the information.The primary articles’ methodological quality was assessed using the Cochrane Collaboration tool to assess the risk of bias (the Cochrane Handbook for Systematic Reviews of Interventions) . The too2 statistic and classified as low (I2 < 25%), moderate (I2 = 25–50%), or high (I2 > 50%) [A narrative synthesis was used to summarise the characteristics of studies, including population and intervention characteristics. Summaries of the association between the outcomes for each study in terms of mean differences (MDs) or standard mean differences (SMDs) using Review Manager 5  were reported. Absolute values and obtained combined measures of the effect of each primary outcome through meta-analysis with a random-effect model due to the expected heterogeneity between studies were compared . Statist2 > 50%) . A sub-a2 > 50%) . Disagre2 > 50%) .The initial search yielded 2599 potential studies. In total, 376 duplicate records were deleted. We screened 2223 titles and abstracts and excluded 2169 records that did not meet the inclusion criteria. Fifty-four of these were assessed as full text. Of these, 30 were excluded for including people under 65 years old, 14 for being conference abstracts, 3 for not being RCTs, 3 for not including exercise as intervention, and 1 for duplicated data. Ultimately, 13 studies met the criteria for eligibility and were included in the review ,42,43,44Three studies were conducted in Taiwan ,41,42, t2 . Sample sizes varied between 21  and 16352 . Most ofFive articles used interventions based on yoga ,38,39,43All studies had a high or unclear risk of bias in at least one domain. The majority of studies claimed to be randomised. However, only half of them explain how the randomisation was conducted ,40,41,43n = 1) [p = 0.0002) (2 = 95%). If we analysed only the studies with PSQI [p < 0.0001) (2 = 98%). The certainty of evidence, according to the GRADE methodology, was moderate. Although four studies [Sleep Quality: A total of 13 studies reported sleep quality ,42,43,44n = 1) , and an n = 1) ,37. A men = 1) ,41,43,44 0.0002) . The hetith PSQI ,40,41,43 0.0001) . The het studies ,37,39,42 studies ,39,42.p = 0.003) with a high heterogeneity (I2 = 64%). However, the programs carried out between 12 weeks and 12 months  and more than 12 months  did not improve sleep quality. The heterogeneity was high in both cases  (p = 0.02) with a high heterogeneity (I2 = 97%), and the programs of more than 12 months  did not improve sleep quality  . If we a quality .p = 0.06). However, the programs carried out in the facilities showed a significant improvement in sleep quality   . No sign2 = 38%). Older people in the IG had no change after intervention  in comparison to CG (p = 0.10). The heterogeneity of the comparison was high (I2 = 82%) (p < 0.0001) (2 = 0%). The certainty of evidence, according to the GRADE methodology, was low.Sleep efficiency: Five studies examined sleep efficiency ,37,38,422 = 82%) . If we a 0.0001) . The hetSleepiness: Only one study examined the Epworth sleepiness scale (ESS) post-intervention . No signThis systematic review provides new information about the effects of exercise on sleep quality measured exclusively in the older adult population, showing that exercise improves sleep quality, particularly in facilities programs, and improves sleep efficiency measured with objective instruments. However, these results must be analysed with caution because there is a high heterogeneity in interventions and baseline characteristics of the population.Our meta-analysis reports that exercise improves sleep quality. One possible explanation could be that physical activity regulates sleep patterns . With inPoor sleep quality is closely related to mental health problems in older adults. Although this research did not explore anxiety or depression as an outcome, exercise in older adults has already been reported to have a positive effect in reducing depression and anxiety . AnotherThe mechanisms that explain the improvement in sleep quality due to exercise in older adults are still not completely clear. One possible explanation is that exercise training provides a benefit on sleep quality by decreasing the sleep latency and the use of sleep medication in older adults . AnotherSleep quality is a concept that includes quantitative aspects of sleep and more subjective aspects, such as “depth” or “restfulness” of sleep . The mosSleep is a complex phenomenon, and the literature suggests that objective and subjective indices measure different components of sleep in older adults . SpecifiOne of the outcomes analysed was sleep efficiency, which was objectively reported through accelerometry and actigraphy ,37 and sMeta-analyses showed very high heterogeneity. This is probably because the populations are very different and range from healthy subjects to older adults with pathologies. However, one aspect that was even more heterogeneous was the intervention. Although most of the programs had a similar frequency (three to five times per week), the duration was highly variable, with programs that lasted between 8 and 12 weeks ,43,44, uThe most common interventions included were yoga ,38,39,43When analysing by setting, we found that home-based programs (defined as at least 50% of their completion at home) were not effective in improving sleep quality ,38,40, wAn important point to consider is the type of physical training. Although most of the studies comply with the principles of a structured exercise program, with frequency, intensity, time, and duration, as usually happens, physical exercise programs are confused with those that encourage physical activity that consists of increasing moderate to vigorous physical activity (MVPA) . These pThis study shows that exercise has effects on sleep quality. Clinicians and decision makers can review these data and carry out exercise programs focused on older adults who report sleep problems with or without pathologies and who report the use of medications for the management of sleep disorders or to improve mental health. This may be particularly relevant at community care levels, such as nursing homes or primary care. On the other hand, it is recommended to carry out supervised programs that comply with the principles of training to achieve the best possible effect on the population.Our study has some limitations. Most of the studies included populations with different diseases or comorbidities, which may limit the extrapolation of the results and recommendations to the entire spectrum of older people, even though our results were statistically significant. The heterogeneous nature of populations implies that many subjects have different pathophysiological behaviours and conditions, a broad spectrum of severity, and the implication or impact that suffering from associated comorbidities may influence the magnitude of reported results. On the other hand, we tried to sub-analyse by health status; however, it was very complex since the definition of healthy is not uniform among the different authors and, therefore, it loses reliability. Additionally, the studies did not report adherence, which is a fundamental factor in determining the effectiveness of an intervention. Future studies must describe the adherence to exercise, since this factor must be analysed together with the results when evaluating the effectiveness of an intervention.Physical exercise programs in older adults improve sleep quality, particularly in facilities, and sleep efficiency measured with objective instruments.