Objective:To analyze the level and clinical significance of IL-18 and IL-18-binding protein (BP) in the bone marrow of patients with myelodysplastic syndrome (MDS) .Methods:A total of 43 newly diagnosed patients with MDS who were admitted to the Department of Hematology, Tianjin Medical University General Hospital, from July 2020 to February 2021 were randomly selected. The control group consisted of 14 patients with acute myeloid leukemia (AML) and 25 patients with iron-deficiency anemia (IDA). The levels of IL-18 and IL-18 BP in the bone marrow supernatant were measured, and their correlations with MDS severity, as well as the functionality of CD8 + T cells and natural killer cells, was analyzed. Results:The levels of IL-18, IL-18 BP, and free IL-18 (fIL-18) in the bone marrow supernatant of patients with MDS were higher than in the IDA group. The level of fIL-18 was linearly and negatively correlated with the MDS-International Prognostic Scoring System (IPSS) score. IL-18 receptor (IL-18Rα) expression on CD8 + T cells in the MDS group was lower than in the IDA group, and the levels of fIL-18 and IL-18Rα were positively correlated with CD8 + T-cell function in the MDS group. Conclusion:IL-18 BP antagonizes IL-18, leading to a decrease in fIL-18 in the bone marrow microenvironment of patients with MDS, affecting CD8 + T-cell function, which is closely related to MDS severity; therefore, it may become a new target for MDS treatment.

Objective To compare the differences in dosimetric parameters between gradient optimization(GO)method and dynamic multi leaf collimator automatic optimization(dMLC-AO)method in craniospinal irradiation(CSI)to provide references for enhancing the feasibility of CSI planning of Monaco treatment planning system.Methods Fifteen patients undergoing CSI at some hospital were retrospectively selected,and a GO plan(enrolled into a GO group)and a dMLC-AO plan(enrolled into a dMLC-AO group)were designed for volumetric modulated arc therapy for each patient by the Monaco treatment planning system.The two groups were compared in terms of the dosimetric parameters of the planning target volume,dose transition zone and organ at risk.SPSS 24.0 statistical software was used for data analysis.Results The dMLC-AO group behaved slightly better than the GO group in D98％ and homogeneity index,with the differences being significant(P＜0.05),while the two groups were not different statistically in D2％,D50％and conformity index(P＞0.05).There were no significant differences between the two groups in radiation doses to the organs at risk except intestinal Dmean(P＞0.05).The GO group gained advantages over the dMLC-AO group in dose distribution conformity at the drop zone,and the target volume length covered by the 36 Gy isodose line for the intermdediate-section plan of the GO group was significantly longer than that of the dMLC-AO group.Conclusion GO method behaves better than dMLC-AO method in controlling effectively the dose drop at the dose transition zone of CSI plan.[Chinese Medical Equipment Journal,2024,45(9):62-66]

Spinal cord stimulation (SCS)-induced analgesia was characterized, and its underlying mechanisms were examined in a spared nerve injury model of neuropathic pain in rats. The analgesic effect of SCS with moderate mechanical hypersensitivity was increased with increasing stimulation intensity between the 20% and 80% motor thresholds. Various frequencies (2, 15, 50, 100, 10000 Hz, and 2/100 Hz dense-dispersed) of SCS were similarly effective. SCS-induced analgesia was maintained without tolerance within 24 h of continuous stimulation. SCS at 2 Hz significantly increased methionine enkephalin content in the cerebrospinal fluid. The analgesic effect of 2 Hz was abolished by μ or κ opioid receptor antagonist. The effect of 100 Hz was prevented by a κ antagonist, and that of 10 kHz was blocked by any of the μ, δ, or κ receptor antagonists, suggesting that the analgesic effect of SCS at different frequencies is mediated by different endorphins and opioid receptors.
Analgesics ; Animals ; Narcotic Antagonists/pharmacology* ; Neuralgia/therapy* ; Opioid Peptides ; Rats ; Receptors, Opioid/physiology* ; Receptors, Opioid, kappa ; Spinal Cord ; Spinal Cord Stimulation

Objective:To investigate the effect of decreased TCOF1 mRNA level on apoptosis of human embryonic stem cell (ESC) line H9-derived neural crest cells (H9-NCC), and to elucidate the post-transcriptional regulation of TCOF1 expression by microRNAs (miRNAs), as well as the role and mechanism of the teratogenic factor retinoic acid (RA) in regulating TCOF1 expression. Methods:Human H9 ESC was induced into H9-NCC and the surface markers of neural crest cells (NCCs), which included P75, HNK-1 and AP2, were detected by immunofluorescence staining and flow cytometry. The expressions of NCCs marker genes SOX9, ZIC1, AP2, P75, PAX3 and SOX10 were detected by real-time quantitative PCR (RT-qPCR). TCOF1 targeting lentiviral RNA interference vector (sh- TCOF1) was used to knock out TCOF1 in H9-NCC, and apoptosis level was measured. Negative control group was set up to observe apoptosis. MiR-654-5p was predicted as the miRNA that may bind to TCOF1 mRNA with bioinformatics analysis. The effects of miRNA mimics on TCOF1 mRNA levels was compared with negative control by RT-qPCR results. The expression of TCOF1 mRNA was detected by treating H9-NCC with 0.1 μmol/L RA, an important metabolite during embryogenesis. The upstream transcription factor of TCOF1 was predicted, which contained HOXA3. HOXA3 knockdown with small interference RNA and TCOF1 mRNA change with RT-qPCR was used to explore the regulatory mechanism of TCOF1 transcription by RA signaling. Results:(1) H9-NCC cell line was successfully induced. Immunofluorescence staining, flow cytometry analysis and RT-qPCR result showed that the induced H9-NCC expressed specific markers of NCCs. (2)Compared with the negative control group, sh- TCOF1 could knock down TCOF1 expression level in H9-NCC (sh- TCOF1 vs. sh-Scramble, P=0.029). TCOF1 knockdown could increase the level of late apoptosis (sh- TCOF1 vs. sh-Scramble, P=0.029) and up-regulate the expression of caspase-3 (sh- TCOF1 vs. sh-Scramble, P<0.001). (3) Compared with the negative control group, miR-654-5p mimic decreased TCOF1 mRNA level in H9-NCC (miR-654-5p vs. miR-Ctrl, P=0.019). (4) Compared with the negative control group, the expression of TCOF1 mRNA in H9-NCC was up-regulated after RA treatment (RA vs. Ctrl, P=0.041); HOXA3 knockdown inhibited the RA induced TCOF1 mRNA up-regulation in H9-NCC (siHOXA3 vs. siCtrl, P=0.008). Conclusions:The decreased expression of TCOF1 induces H9-NCC apoptosis, miR-654-5p down-regulates TCOF1 mRNA through post-transcriptional regulation, and RA promotes TCOF1 expression through HOXA3 mediated pathway.

Objective:To investigate the effect of decreased TCOF1 mRNA level on apoptosis of human embryonic stem cell (ESC) line H9-derived neural crest cells (H9-NCC), and to elucidate the post-transcriptional regulation of TCOF1 expression by microRNAs (miRNAs), as well as the role and mechanism of the teratogenic factor retinoic acid (RA) in regulating TCOF1 expression. Methods:Human H9 ESC was induced into H9-NCC and the surface markers of neural crest cells (NCCs), which included P75, HNK-1 and AP2, were detected by immunofluorescence staining and flow cytometry. The expressions of NCCs marker genes SOX9, ZIC1, AP2, P75, PAX3 and SOX10 were detected by real-time quantitative PCR (RT-qPCR). TCOF1 targeting lentiviral RNA interference vector (sh- TCOF1) was used to knock out TCOF1 in H9-NCC, and apoptosis level was measured. Negative control group was set up to observe apoptosis. MiR-654-5p was predicted as the miRNA that may bind to TCOF1 mRNA with bioinformatics analysis. The effects of miRNA mimics on TCOF1 mRNA levels was compared with negative control by RT-qPCR results. The expression of TCOF1 mRNA was detected by treating H9-NCC with 0.1 μmol/L RA, an important metabolite during embryogenesis. The upstream transcription factor of TCOF1 was predicted, which contained HOXA3. HOXA3 knockdown with small interference RNA and TCOF1 mRNA change with RT-qPCR was used to explore the regulatory mechanism of TCOF1 transcription by RA signaling. Results:(1) H9-NCC cell line was successfully induced. Immunofluorescence staining, flow cytometry analysis and RT-qPCR result showed that the induced H9-NCC expressed specific markers of NCCs. (2)Compared with the negative control group, sh- TCOF1 could knock down TCOF1 expression level in H9-NCC (sh- TCOF1 vs. sh-Scramble, P=0.029). TCOF1 knockdown could increase the level of late apoptosis (sh- TCOF1 vs. sh-Scramble, P=0.029) and up-regulate the expression of caspase-3 (sh- TCOF1 vs. sh-Scramble, P<0.001). (3) Compared with the negative control group, miR-654-5p mimic decreased TCOF1 mRNA level in H9-NCC (miR-654-5p vs. miR-Ctrl, P=0.019). (4) Compared with the negative control group, the expression of TCOF1 mRNA in H9-NCC was up-regulated after RA treatment (RA vs. Ctrl, P=0.041); HOXA3 knockdown inhibited the RA induced TCOF1 mRNA up-regulation in H9-NCC (siHOXA3 vs. siCtrl, P=0.008). Conclusions:The decreased expression of TCOF1 induces H9-NCC apoptosis, miR-654-5p down-regulates TCOF1 mRNA through post-transcriptional regulation, and RA promotes TCOF1 expression through HOXA3 mediated pathway.

Berberine(BBR)is an isoquinoline alkaloid extracted from Coptis chinensis that improves diabetes,hyperlipidemia and inflammation.Due to the low oral bioavailability of BBR,its mechanism of action is closely related to the gut microbiota.This study focused on the CYP51 enzyme of intestinal bacteria to elucidate a new mechanism of BBR transformation by demethylation in the gut microbiota through multiple analytical techniques.First,the docking of BBR and CYP51 was performed;then,the pharma-cokinetics of BBR was determined in ICR mice in vivo,and the metabolism of BBR in the liver,kidney,gut microbiota and single bacterial strains was examined in vitro.Moreover,16S rRNA analysis of ICR mouse feces indicated the relationship between BBR and the gut microbiota.Finally,recombinant E.coli con-taining cyp51 gene was constructed and the CYP51 enzyme lysate was induced to express.The metabolic characteristics of BBR were analyzed in the CYP51 enzyme lysate system.The results showed that CYP51 in the gut microbiota could bind stably with BBR,and the addition of voriconazole(a specific inhibitor of CYP51)slowed down the metabolism of BBR,which prevented the production of the demethylated metabolites thalifendine and berberrubine.This study demonstrated that CYP51 promoted the deme-thylation of BBR and enhanced its intestinal absorption,providing a new method for studying the metabolic transformation mechanism of isoquinoline alkaloids in vivo.

Objective: To determine whether T1 mapping could monitor the dynamic changes of injury in myocardial infarction (MI) and be histologically validated. Materials and Methods: In 22 pigs, MI was induced by ligating the left anterior descending artery and they underwent serial cardiovascular magnetic resonance examinations with modified Look-Locker inversion T1 mapping and extracellular volume (ECV) computation in acute (within 24 hours, n = 22), subacute (7 days, n = 13), and chronic (3 months, n = 7) phases of MI. Masson’s trichrome staining was performed for histological ECV calculation. Myocardial native T1 and ECV were obtained by region of interest measurement in infarcted, peri-infarct, and remote myocardium. Results: Native T1 and ECV in peri-infarct myocardium differed from remote myocardium in acute (1181 ± 62 ms vs. 1113 ± 64 ms, p = 0.002; 24 ± 4% vs. 19 ± 4%, p = 0.031) and subacute phases (1264 ± 41 ms vs. 1171 ± 56 ms, p < 0.001; 27 ± 4% vs. 22 ± 2%, p = 0.009) but not in chronic phase (1157 ± 57 ms vs. 1120 ± 54 ms, p = 0.934; 23 ± 2% vs. 20 ± 1%, p = 0.109). From acute to chronic MI, infarcted native T1 peaked in subacute phase (1275 ± 63 ms vs. 1637 ± 123 ms vs. 1471 ± 98 ms, p < 0.001), while ECV progressively increased with time (35 ± 7% vs. 46 ± 6% vs. 52 ± 4%,p < 0.001). Native T1 correlated well with histological findings (R2 = 0.65 to 0.89, all p < 0.001) so did ECV (R2 = 0.73 to 0.94, all p < 0.001). Conclusion T1 mapping allows the quantitative assessment of injury in MI and the noninvasive monitoring of tissue injury evolution, which correlates well with histological findings.

Objective: To determine whether T1 mapping could monitor the dynamic changes of injury in myocardial infarction (MI) and be histologically validated. Materials and Methods: In 22 pigs, MI was induced by ligating the left anterior descending artery and they underwent serial cardiovascular magnetic resonance examinations with modified Look-Locker inversion T1 mapping and extracellular volume (ECV) computation in acute (within 24 hours, n = 22), subacute (7 days, n = 13), and chronic (3 months, n = 7) phases of MI. Masson’s trichrome staining was performed for histological ECV calculation. Myocardial native T1 and ECV were obtained by region of interest measurement in infarcted, peri-infarct, and remote myocardium. Results: Native T1 and ECV in peri-infarct myocardium differed from remote myocardium in acute (1181 ± 62 ms vs. 1113 ± 64 ms, p = 0.002; 24 ± 4% vs. 19 ± 4%, p = 0.031) and subacute phases (1264 ± 41 ms vs. 1171 ± 56 ms, p < 0.001; 27 ± 4% vs. 22 ± 2%, p = 0.009) but not in chronic phase (1157 ± 57 ms vs. 1120 ± 54 ms, p = 0.934; 23 ± 2% vs. 20 ± 1%, p = 0.109). From acute to chronic MI, infarcted native T1 peaked in subacute phase (1275 ± 63 ms vs. 1637 ± 123 ms vs. 1471 ± 98 ms, p < 0.001), while ECV progressively increased with time (35 ± 7% vs. 46 ± 6% vs. 52 ± 4%,p < 0.001). Native T1 correlated well with histological findings (R2 = 0.65 to 0.89, all p < 0.001) so did ECV (R2 = 0.73 to 0.94, all p < 0.001). Conclusion T1 mapping allows the quantitative assessment of injury in MI and the noninvasive monitoring of tissue injury evolution, which correlates well with histological findings.

The pain-relieving effect of acupuncture is known to involve primary afferent nerves (PANs) via their roles in signal transmission to the CNS. Using single-unit recording in rats, we characterized the generation and transmission of electrical signals in Aβ and Aδ fibers induced by acupuncture-like stimuli. Acupuncture-like signals were elicited in PANs using three techniques: manual acupuncture (MAc), emulated acupuncture (EAc), and electro-acupuncture (EA)-like peripheral electrical stimulation (PES). The discharges evoked by MAc and EAc were mostly in a burst pattern with average intra-burst and inter-burst firing rates of 90 Hz and 2 Hz, respectively. The frequency of discharges in PANs was correlated with the frequency of PES. The highest discharge frequency was 246 Hz in Aβ fibers and 180 Hz in Aδ fibers. Therefore, EA in a dense-disperse mode (at alternating frequency between 2 Hz and 15 Hz or between 2 Hz and 100 Hz) best mimics MAc. Frequencies of EA output >250 Hz appear to be obsolete for pain relief.

Objective To develop chitosan composite keratinocyte growth factor-2 mutant(KGF-2M)temperature-sen-sitive dressing and evaluate its physicochemical properties and dynamic release rule were used.Methods Chitosan, chi-tosan quaternary ammonium salt,β-glycerophosphate and other adjuvant materials to configure different formulations which were compounded with KGF-2M in order to develop temperature-sensitive dressing.Gelling time, temperature,the release rate of KGF-2M and other indicators were measured to analyze the physical and chemical properties of the temperature -sen-sitive dressing.Results Chitosan-KGF-2M composite dressing with temperature-sensitive properties was obtained by opti-mizing the formulation components of chitosan and related adjuvant materials.When the liquid dressing was above 35℃,it could be converted from liquid to solid gelatin within 10 minutes.The compound KGF-2M released from the gel was more than 98%at 4 h,and its bioactivity remained stable.Conclusion The thermo-sensitive gel has the characteristics of good conformability,moisturizing(moisture),isolation,wound healing,and a controlled release effect,which has great potential in wartime for wound repair.