OBJECTIVE: To investigate the effects of removing microglia from spinal cord on nerve repair and functional recovery after spinal cord injury (SCI) in mice. METHODS: Thirty-nine 6-week-old female C57BL/6 mice were randomly divided into control group ( n=12), SCI group ( n=12), and PLX3397+SCI group ( n=15). The PLX3397+SCI group received continuous feeding of PLX3397, a colony-stimulating factor 1 receptor inhibitor, while the other two groups were fed a standard diet. After 14 days, both the SCI group and the PLX3397+SCI group were tested for ionized calcium binding adapter molecule 1 (Iba1) to confirm that the PLX3397+SCI group had completely depleted the spinal cord microglia. The SCI model was then prepared by clamping the spinal cord in both the SCI group and the PLX3397+SCI group, while the control group underwent laminectomy. Preoperatively and at 1, 3, 7, 14, 21, and 28 days postoperatively, the Basso Mouse Scale (BMS) was used to assess the hind limb function of mice in each group. At 28 days, a footprint test was conducted to observe the gait of the mice. After SCI, spinal cord tissue from the injury site was taken, and Iba1 immunofluorescence staining was performed at 7 days to observe the aggregation and proliferation of microglia in the spinal cord. HE staining was used to observe the formation of glial scars at the injury site at 28 days; glial fibrillary acidic protein (GFAP) immunofluorescence staining was applied to astrocytes to assess the extent of the injured area; neuronal nuclei antigen (NeuN) immunofluorescence staining was used to evaluate neuronal survival. And 5-hydroxytryptamine (5-HT) immunofluorescence staining was performed to assess axonal survival at 60 days. RESULTS: All mice survived until the end of the experiment. Immunofluorescence staining revealed that the microglia in the spinal cord of the PLX3397+SCI group decreased by more than 95% compared to the control group after 14 days of continuous feeding with PLX3397 ( P<0.05). Compared to the control group, the BMS scores in the PLX3397+SCI group and the SCI group significantly decreased at different time points after SCI ( P<0.05). Moreover, the PLX3397+SCI group showed a further decrease in BMS scores compared to the SCI group, and exhibited a dragging gait. The differences between the two groups were significant at 14, 21, and 28 days ( P<0.05). HE staining at 28 days revealed that the SCI group had formed a well-defined and dense gliotic scar, while the PLX3397+SCI group also developed a gliotic scar, but with a more blurred and loose boundary. Immunofluorescence staining revealed that the number of microglia near the injury center at 7 days increased in the SCI group than in the control group, but the difference between groups was not significant ( P>0.05). In contrast, the PLX3397+SCI group showed a significant reduction in microglia compared to both the control and SCI groups ( P<0.05). At 28 days after SCI, the area of spinal cord injury in the PLX3397+SCI group was significantly larger than that in SCI group ( P<0.05); the surviving neurons significantly reduced compared with the control group and SCI group ( P<0.05). The axonal necrosis and retraction at 60 days after SCI were more obvious. CONCLUSION The removal of microglia in the spinal cord aggravate the tissue damage after SCI and affecte the recovery of motor function in mice, suggesting that microglia played a neuroprotective role in SCI.
Animals ; Spinal Cord Injuries/surgery* ; Microglia/pathology* ; Female ; Mice ; Mice, Inbred C57BL ; Nerve Regeneration/drug effects* ; Spinal Cord/pathology* ; Pyrroles/administration & dosage* ; Aminopyridines/administration & dosage* ; Recovery of Function ; Disease Models, Animal ; Calcium-Binding Proteins/metabolism* ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor/antagonists & inhibitors* ; Microfilament Proteins/metabolism* ; Glial Fibrillary Acidic Protein/metabolism*

Objectives:To explore whether short-course radiotherapy (SCRT)-based total neoadjuvant therapy (TNT) combined with PD-1 inhibitors could further promote tumor regression and improve the prognosis.Methods:This is a prospective, multicenter, two-arm randomized controlled, seamless phase Ⅱ/Ⅲ trial for proficient mismatch repair or microsatellite stable (pMMR/MSS) locally advanced rectal cancer (LARC). Eligible patients were randomly assigned to the iTNT (TNT+PD-1) group or the TNT group. Patients in the TNT group received SCRT (5 Gy×5) followed by 4 cycles of CAPOX or 6 cycles of mFOLFOX chemotherapy, with the iTNT group receiving SCRT followed by the same regime in combination with 4 cycles of Sintilimab. Total mesorectal excision (TME) surgery or watch and wait (W&W) was performed after neoadjuvant therapy and then 2 cycles of same regimen as before were recommended. The primary endpoints are the complete response (CR) rate for phase Ⅱ trial and 3-year disease-free survival (DFS) for phase Ⅲ trial. A total of 588 patients will be enrolled for the phase Ⅱ/Ⅲ trial. Short-term efficacy and safety data from the initial 100 treated patients were analyzed as planned.Results:From 2022-8-31 to 2023-5-24 the initial 100 patients were enrolled from 10 hospitals in China, 76.0%(76/100) patients were male, and the median age was 61 years (21-74 years). More patients had tumors located in the lower rectum (78.0%, 78/100), staged T3-4 (97.0%, 97/100) and N1-2 (93.0%, 93/100), and about half of the tumors invaded the mesorectal fascia (52.0%, 52/100) and with extramural vascular invasion (51.0%, 51/100). Analyses were performed according to the per-protocal (PP) set. All patients in the iTNT group ( n=52) and the TNT group ( n=48) completed SCRT; The 4-cycle chemotherapy±Sintilimab completion rates were 86.5% and 100.0% in the iTNT and TNT groups, respectively. In the iTNT group, 82.7% (43/52), 11.5% (6/52), and 5.8% (3/52) of the patients received 4, 3, and 2 cycles of PD-1 inhibitor. After TNT, 68 patients underwent radical surgery and 15 patients achieved cCR and adopted W&W. The pathological complete response (pCR) rates were 48.5% (16/33) and 17.1% (6/35) in the iTNT and TNT groups, with CR rates of 50.0% (25/50) and 26.1% (12/46), respectively. The incidence of treatment-related grade 3-4 adverse events was 26.9% (14/52, iTNT group) and 18.8% (9/48, TNT group), with thrombocytopenia and leukopenia being the most common. Among patients receiving immunotherapy, grade 3 immunotherapy-related adverse events occurred in 2 (3.8%, 2/52) patients: one case was pancreatitis, another case was hepatitis combined with myositis and myocarditis. Conclusion:The preliminary results show that SCRT-based TNT combined with PD-1 inhibitors could further improve the CR rate for LARC without unexpected serious adverse events.

Objective To investigate the efficacy of surface electromyography biofeedback combined with respiratory training on post-stroke dysphagia and its impacts on serum neuron specific enolase(NSE)and insulin-like growth factor 1(IGF-1)in patients.Methods Totally 120 patients with post-stroke dysphagia in our hospital were stochastically assigned into the control group and the combined group,with 60 patients in each group.Both groups were given conventional treatment first,while the control group received respiratory training treatment.The combined group received surface electromyography biofeedback treatment on the top of the control group.The efficacy,serum NSE,central nervous system specific protein(S100β),IGF-1,functional oral intake scale(FOIS)score,standard swallowing function assessment scale(SSA)score,respiratory function,pharyngeal contraction rate and duration of pharyngeal contraction were compared between the two groups.Results After treatment,the total effective rate was higher in the combined group(x2=4.876,P＜0.05).Compared with before treatment,NSE,S100β,SSA score and pharyngeal contraction rate decreased after treatment in both groups,and those were even lower in the combined group(t=5.193,9.000,8.976,10.614,P＜0.05).Compared with before treatment,IGF-1,FOIS score,force vital capacity(FVC),forced expiratory volume in one second(FEV1),peak expiratory flow rate(PEF)and duration of pharyngeal contraction increased after treatment in both groups,with even higher levels in the combined group(t=4.212,6.220,3.765,6.935,5.020,4.249,P＜0.05).Conclusion Surface electromyography biofeedback combined with respiratory training can improve neuronal damage in patients with dysphagia,enhance swallowing efficiency,reduce serum NSE,S100β levels and pharyngeal retention time,with significant curative effect.

Objective To investigate the incidence of functional bowel disease(FBD)and its relationship with depression,anxiety and sleep in navy submarine forces.Methods A questionnaire survey on the incidence of FBD was conducted on 364 naval soldiers who were enrolled according to Rome Ⅳ classification and diagnostic criteria.The risk factors of FBD were analyzed.The incidence of specific diseases of FBD was compared among soldiers with different jobs.The depression,anxiety,and sleep quality were investigated in FBD patients.Results In the 364 participants,132(36.3%)were diagnosed with FBD according to Roman Ⅳcriteria.Military rank and marital status might be risk factors for FBD.There was no significant difference in the overall incidence of FBD among the participants with various jobs(P>0.05).The incidence of functional constipation in the submariners was significantly higher than that in the land soldiers,while the incidence of functional diarrhea in the land soldiers was higher than that in the submariners(all P<0.05).The incidences of depression,anxiety and sleep disorder in the participants with FBD were higher than those in the participants without FBD.Conclusion There is a high incidence of FBD in submariners,which may be related to their psychological states.

Objective To explore the mechanism of Luzhongjiangu Decoction in treating Osteonecrosis of the femoral head(ONFH).Methods Animal experiments were conducted to validate the therapeutic effect of Luzhongjiangu Decoction on ONFH.Network pharmacology was employed to screen potential targets,and molecular docking was used to verify the affinity between ligands and receptors.Fifty rats were randomly divided into blank group,model group,high-dose Luzhongjiangu Decoction groups,medium-dose Luzhongjiangu Decoction groups and low-dose Luzhongjiangu Decoction groups.After treatment,the therapeutic effect was evaluated by bone histomorphometry,bone histopathology and the expression of osteogenic related proteins.The serum metabolic differences were detected by UHPLC-QE-MS non-targeted metabonomics technique,followed by metabolic pathway enrichment analysis.Network pharmacology and metabolomics were integrated,and molecular docking was employed to further investigate the underlying mechanisms..Results The experimental results showed that Luzhongjiangu Decoction can significantly increase the expression of OCN,OPN and other osteogenic related factors,increase bone mineral density,alleviate the destruction of bone trabeculae,compared with that in the model group(P<0.01).The metabolomics test showed that compared with model group,Luzhongjiangu Decoction could callback 34 kinds of serum metabolic differences,involving 21 metabolic pathways,the main metabolic pathways were arachidonic acid metabolism,steroid hormone biosynthesis,primary bile acid biosynthesis,lysine degradation and so on.Network pharmacological results suggested that 185 active components and 1040 targets,551 disease targets and 76 intersection targets were obtained.The key pathways of Luzhongjiangu Decoction in the treatment of ONFH,which were analyzed by KEGG,involved HIF-1 signaling pathway,thyroid hormone signaling pathway,Calcium signaling pathway and so on.Integrated analysis showed that Luzhongjiangu Decoction may treat ONFH through seven metabolic pathways including arginine and proline metabolism,arachidonic acid metabolism,and steroid hormone biosynthesis;the molecular docking results also revealed correlation between potential pharmacodynamic substances and core target genes.Conclusion Luzhongjiangu Decoction can regulate the balance between proliferation and apoptosis of bone cells in a hypoxic environment by affecting the metabolic pathways such as arachidonic acid metabolism and HIF-1 signaling pathway,thereby exerting a therapeutic effect on osteonecrosis of the femoral head.

Objectives:To explore whether short-course radiotherapy (SCRT)-based total neoadjuvant therapy (TNT) combined with PD-1 inhibitors could further promote tumor regression and improve the prognosis.Methods:This is a prospective, multicenter, two-arm randomized controlled, seamless phase Ⅱ/Ⅲ trial for proficient mismatch repair or microsatellite stable (pMMR/MSS) locally advanced rectal cancer (LARC). Eligible patients were randomly assigned to the iTNT (TNT+PD-1) group or the TNT group. Patients in the TNT group received SCRT (5 Gy×5) followed by 4 cycles of CAPOX or 6 cycles of mFOLFOX chemotherapy, with the iTNT group receiving SCRT followed by the same regime in combination with 4 cycles of Sintilimab. Total mesorectal excision (TME) surgery or watch and wait (W&W) was performed after neoadjuvant therapy and then 2 cycles of same regimen as before were recommended. The primary endpoints are the complete response (CR) rate for phase Ⅱ trial and 3-year disease-free survival (DFS) for phase Ⅲ trial. A total of 588 patients will be enrolled for the phase Ⅱ/Ⅲ trial. Short-term efficacy and safety data from the initial 100 treated patients were analyzed as planned.Results:From 2022-8-31 to 2023-5-24 the initial 100 patients were enrolled from 10 hospitals in China, 76.0%(76/100) patients were male, and the median age was 61 years (21-74 years). More patients had tumors located in the lower rectum (78.0%, 78/100), staged T3-4 (97.0%, 97/100) and N1-2 (93.0%, 93/100), and about half of the tumors invaded the mesorectal fascia (52.0%, 52/100) and with extramural vascular invasion (51.0%, 51/100). Analyses were performed according to the per-protocal (PP) set. All patients in the iTNT group ( n=52) and the TNT group ( n=48) completed SCRT; The 4-cycle chemotherapy±Sintilimab completion rates were 86.5% and 100.0% in the iTNT and TNT groups, respectively. In the iTNT group, 82.7% (43/52), 11.5% (6/52), and 5.8% (3/52) of the patients received 4, 3, and 2 cycles of PD-1 inhibitor. After TNT, 68 patients underwent radical surgery and 15 patients achieved cCR and adopted W&W. The pathological complete response (pCR) rates were 48.5% (16/33) and 17.1% (6/35) in the iTNT and TNT groups, with CR rates of 50.0% (25/50) and 26.1% (12/46), respectively. The incidence of treatment-related grade 3-4 adverse events was 26.9% (14/52, iTNT group) and 18.8% (9/48, TNT group), with thrombocytopenia and leukopenia being the most common. Among patients receiving immunotherapy, grade 3 immunotherapy-related adverse events occurred in 2 (3.8%, 2/52) patients: one case was pancreatitis, another case was hepatitis combined with myositis and myocarditis. Conclusion:The preliminary results show that SCRT-based TNT combined with PD-1 inhibitors could further improve the CR rate for LARC without unexpected serious adverse events.

Objective To investigate the efficacy of surface electromyography biofeedback combined with respiratory training on post-stroke dysphagia and its impacts on serum neuron specific enolase(NSE)and insulin-like growth factor 1(IGF-1)in patients.Methods Totally 120 patients with post-stroke dysphagia in our hospital were stochastically assigned into the control group and the combined group,with 60 patients in each group.Both groups were given conventional treatment first,while the control group received respiratory training treatment.The combined group received surface electromyography biofeedback treatment on the top of the control group.The efficacy,serum NSE,central nervous system specific protein(S100β),IGF-1,functional oral intake scale(FOIS)score,standard swallowing function assessment scale(SSA)score,respiratory function,pharyngeal contraction rate and duration of pharyngeal contraction were compared between the two groups.Results After treatment,the total effective rate was higher in the combined group(x2=4.876,P＜0.05).Compared with before treatment,NSE,S100β,SSA score and pharyngeal contraction rate decreased after treatment in both groups,and those were even lower in the combined group(t=5.193,9.000,8.976,10.614,P＜0.05).Compared with before treatment,IGF-1,FOIS score,force vital capacity(FVC),forced expiratory volume in one second(FEV1),peak expiratory flow rate(PEF)and duration of pharyngeal contraction increased after treatment in both groups,with even higher levels in the combined group(t=4.212,6.220,3.765,6.935,5.020,4.249,P＜0.05).Conclusion Surface electromyography biofeedback combined with respiratory training can improve neuronal damage in patients with dysphagia,enhance swallowing efficiency,reduce serum NSE,S100β levels and pharyngeal retention time,with significant curative effect.

Objective To explore the mechanism of Luzhongjiangu Decoction in treating Osteonecrosis of the femoral head(ONFH).Methods Animal experiments were conducted to validate the therapeutic effect of Luzhongjiangu Decoction on ONFH.Network pharmacology was employed to screen potential targets,and molecular docking was used to verify the affinity between ligands and receptors.Fifty rats were randomly divided into blank group,model group,high-dose Luzhongjiangu Decoction groups,medium-dose Luzhongjiangu Decoction groups and low-dose Luzhongjiangu Decoction groups.After treatment,the therapeutic effect was evaluated by bone histomorphometry,bone histopathology and the expression of osteogenic related proteins.The serum metabolic differences were detected by UHPLC-QE-MS non-targeted metabonomics technique,followed by metabolic pathway enrichment analysis.Network pharmacology and metabolomics were integrated,and molecular docking was employed to further investigate the underlying mechanisms..Results The experimental results showed that Luzhongjiangu Decoction can significantly increase the expression of OCN,OPN and other osteogenic related factors,increase bone mineral density,alleviate the destruction of bone trabeculae,compared with that in the model group(P<0.01).The metabolomics test showed that compared with model group,Luzhongjiangu Decoction could callback 34 kinds of serum metabolic differences,involving 21 metabolic pathways,the main metabolic pathways were arachidonic acid metabolism,steroid hormone biosynthesis,primary bile acid biosynthesis,lysine degradation and so on.Network pharmacological results suggested that 185 active components and 1040 targets,551 disease targets and 76 intersection targets were obtained.The key pathways of Luzhongjiangu Decoction in the treatment of ONFH,which were analyzed by KEGG,involved HIF-1 signaling pathway,thyroid hormone signaling pathway,Calcium signaling pathway and so on.Integrated analysis showed that Luzhongjiangu Decoction may treat ONFH through seven metabolic pathways including arginine and proline metabolism,arachidonic acid metabolism,and steroid hormone biosynthesis;the molecular docking results also revealed correlation between potential pharmacodynamic substances and core target genes.Conclusion Luzhongjiangu Decoction can regulate the balance between proliferation and apoptosis of bone cells in a hypoxic environment by affecting the metabolic pathways such as arachidonic acid metabolism and HIF-1 signaling pathway,thereby exerting a therapeutic effect on osteonecrosis of the femoral head.

Objective:To compare the assay and related substance detection methods of erythromycin lactobionate in the Chinese Pharmacopoeia 2020(ChP),USP 2023(USP),JP18(JP),BP2023(BP)and EP 11.0(EP),investigate the differences between the test results obtained from 7 batches of erythromycin lactobionate for injection samples by using ChP and BP methods and thus provide a reference for the improvement of specification of erythromycin lactobionate.Methods:The differences of test methods and limits under the items of assay and related substances of erythromycin lactobionate in the above five pharmacopoeias were listed and compared.The related substances and contents of erythromycin lactobionate for injection samples from different manufactures were tested with methods stated in ChP and BP,and then compared and analyzed.Results:The items of related substances were determined by high performance liquid chromotographic methods in the ChP,EP and BP.The test methods and limits in the EP and BP were the same,which were different from that in ChP.The related substances were not determined in the USP and JP.There are obvious differences between the chromatographic methods and limits for the items of related substances in the ChP and BP,i.e.,BP contains 7 specific impuri-ties,while ChP contains only 2 specific impurities.The limits for any other impurity and the total amount of impurities were lower in the BP than those in the ChP.The antibiotic microbiological assay was used as the test method for the item of assay in the ChP,USP and JP based on different bacterial strains,which was different from the chromatographic method used in the BP.Based on the different methods of the ChP and BP,the related substances and content determination results of 7 batches of erythromycin lactobionate for injection samples met the acceptance criteria.The detection efficiency of BP related substances inspection method for specific impurities and total impurities were significantly higher than that of the ChP method.Conclusion:The BP method is superi-or to the ChP method in the detection of erythromycin lactobionate and erythromycin lactobionate for injection related substances.In terms of content determination,the external standard method adopted by BP is feasible to replace the antibiotic microbiological assay adopted by the ChP.Related substances and content determination methods adopted by the BP can provide an important reference for the revision and improvement of erythromycin lactobionate standard in the ChP.

ObjectiveTo investigate the current status of cognition and attitude towards hospice care among medical staff in Liaocheng，analyze the related influencing factors，and to provide reference for further development of hospice care services. MethodsUsing the method of convenient sampling，404 medical staff from all levels of hospitals in Liaocheng were selected as the research subjects from January to June 2022 to conduct a questionnaire survey on the cognition and attitude towards hospice care.Statistical methods were used to analyze the related influencing factors. ResultsThe knowledge score of hospice care among medical staff in Liaocheng was （13.02 ± 4.10），with an average score rate of 65.10%.The score of attitude was （38.67 ± 5.64），with an average score rate of 64.50%.Age （41~50 years old），having received education and training，treated or cared for patients in the middle and late stages，and understanding ethics and morality，as well as cultural customs were significantly positively correlated with knowledge scores.Age （> 50 years old），professional title （deputy senior professional title），position （medical treatment），and experience in treating or caring for patients in the middle and late stages were positively related to attitude scores. ConclusionThe cognition and attitude towards hospice care among medical staff in Liaocheng were at a moderate level.Strengthening the construction of a standardized hospice care system is helpful to improve the cognition and attitude level towards hospice care among medical staff.