OBJECTIVE To form an expert consensus addressing clinical issues regarding the use of parenteral direct thrombin inhibitors （DTIs） in special populations. METHODS Led by the Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital（the Affiliated Hospital of UESTC）， a multidisciplinary working group was formed comprising experts from multiple fields， including clinical pharmacy， cardiac surgery， obstetrics， pediatrics and evidence-based medicine. Through literature review and the Delphi method， clinical questions regarding the efficacy and safety of parenteral DTIs used in special populations were identified. A structured design was adopted using the “Population-Intervention-Comparison-Outcome” （PICO） framework；systematic searches were conducted in CJFD， PubMed， Embase and other databases. Relevant evidence from randomized controlled trials，cohort studies and systematic reviews were included and synthesized. Evidence quality was assessed using the Grading of Recommendations Assessment，Development and Evaluation （GRADE） approach， and recommendations were formulated through three rounds of Delphi surveys and expert consensus meetings. RESULTS &CONCLUSIONS Seven clinical questions were ultimately selected （with a consensus rate exceeding 90%）， resulting in the formulation of seven recommendations on the use of parenteral DTIs in special populations， including children， pregnant women， patients with hepatic or renal impairment， patients with mesenteric venous thrombosis， and individuals with thrombophilia. These recommendations clarify the preferred agents， dosing ranges， monitoring parameters， and safety management strategies for parenteral DTIs in these special populations. This expert consensus， which is formulated based on the best available evidence， provides evidence-based guidance for standardized and individualized use of parenteral DTIs in special populations.

ObjectiveTo develop an assessment tool for apraxia of speech (AOS) of Chinese Putonghua speakers and test its reliability and validity. MethodsThe Chinese Apraxia of Speech Assessment and Diagnostic Tool (CAADT) was developed based on the Apraxia of Speech Rating Scale 3.5, combined with the linguistic characteristics of Chinese and clinical experience. The tool consistsed of eleven items across three sections: articulation, prosody and alternating motion rates. Six experts evaluated the content validity. From November, 2024 to May, 2025, 51 patients with post-stroke AOS (experimental group) and ten patients with post-stroke aphasia without AOS (control group) were recruited from Anhui Wannan Rehabilitation Hospital (the Fifth People's Hospital of Wuhu), and tested with CAADT. Reliability was assessed using Cronbach's α coefficient, Kendall's coefficient of concordance W and Pearson correlation coefficient. Validity was evaluated using the content validity index (CVI) and Spearman correlation coefficient. Discriminative effect was analyzed using the receiver operating characteristic (ROC) curve. ResultsThe Cronbach's α coefficients for the articulation and prosody sections and the total scale were all > 0.9, while it was 0.454 for the alternating motion rates. Inter-rater reliability was good (W ≥ 0.598, P < 0.001). Test-retest reliability showed high positive correlations for the three sections and the total score between the two assessments (r ≥ 0.84, P < 0.001). The scale-level CVI was 0.95, and the item-level CVI ≥ 0.83. The Spearman correlation coefficients among the sections ranged from 0.30 to 0.70. ROC analysis revealed an area under the curve of 0.953, with a cut-off value of 11, yielding a sensitivity of 0.92 and a specificity of 0.90. ConclusionCAADT demonstrates good reliability, validity and discriminative effect, which can be used for clinical assessment and auxiliary diagnosis of Chinese Putonghua speaking patients with post-stroke AOS.

BACKGROUND:As a degradable scaffold material for bone tissue engineering,lactic acid is widely used in tissue regeneration and repair research,and plays an important role in promoting tissue healing,new bone formation and angiogenesis. OBJECTIVE:To observe the effect of lactic acid degradation products on osteoclasts and to investigate the effects of lactic-interfered osteoclast conditioned medium on the proliferation,migration and tube-forming capacity of human umbilical vein endothelial cells. METHODS:(1)The mouse monocyte macrophage cell line RAW264.7 at logarithmic growth period was selected,and adherent cells were cultured in the osteoclast induction medium(DMEM medium with nuclear factor-κB receptor-activating factor ligand and 10%fetal bovine serum)containing different concentrations of lactic acid(0,5,10,20 mmol/L).After 5 days of culture,tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining were conducted.After 24 hours of culture,RT-PCR was used to detect the mRNA expression of tartrate-resistant acid phosphatase 5.(2)RAW264.7 cells at logarithmic growth period were selected and adherent cells were divided into two groups.Control group was cultured in the osteoclast induction medium,while experimental group was cultured in the osteoclast induction medium containing 10 mmol/L lactic acid.After 5 days of culture,the medium in each group was removed and the cells in the two groups were cultured in the serum-free DMEM medium for another 24 hours.Cell supernatant was then collected and used as the conditioned medium after mixed with an equal volume of DMEM medium containing 10%fetal bovine serum.Human umbilical vein endothelial cells at the logarithmic growth phase were taken and separately co-cultured with the conditioned medium of the control and experimental groups.The proliferation,migration and tube-forming ability of human umbilical vein endothelial cells were observed by cell counting kit-8 assay,migration assay,scratch assay and tube-forming assay.The mRNA and protein expression of angiogenesis-related genes and proteins were observed by RT-PCR and western blot. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and cytoskeletal fibrillar actin staining showed that 5 and 10 mmol/L lactic acid promoted osteoclastic differentiation of RAW264.7 cells and the promoting effect of 10 mmol/L lactate was more significant.RT-PCR results showed that the expression of tartrate-resistant acid phosphatase-5 mRNA of osteoclast-related genes was the highest when the lactic acid concentration was 5,10,and 20 mmol/L(P<0.05),especially 10 mmol/L.Compared with the control group,the proliferation,migration and tube-forming abilities of human umbilical vein endothelial cells were significantly increased in the experimental group(P<0.05).Compared with the control group,the expression levels of vascular endothelial growth factor and angiogenin 1 mRNA and protein were increased in the experimental group(P<0.05).To conclude,lactate-induced osteoclast conditioned medium could promote the angiogenesis of endothelial cells,and the mechanism may be related to the promotion of the expression of vascular endothelial growth factor and angiogenin 1.

Aim To study the therapeutic effect of al-licin on senna-induced diarrhea in mice and to explore the underlying mechanism.Methods Forty-eight C57BL/6J mice were randomly divided into six groups:control,model,loperamide positive control group(2 mg·kg-1),allicin low-dose group(6 mg·kg-1),allicin medium-dose group(12 mg·kg-1)and allicin high-dose group(18 mg·kg-1).Except for the con-trol group,the diarrhea model was induced in the other groups by intragastric administration of senna leaf ex-tract.After drug administration,several diarrhea indi-ces were measured:the rate of loose stools,diarrhea index,accumulated frequency of loose stools at differ-ent time points within 5 hours,and small intestine pro-pelling rate.Serum levels of TNF-α and IL-6 were de-tected by ELISA.Serum NO content was determined u-sing the Griess method.The activities of SOD and CAT,as well as MDA content in the ileum and colon,were measured.The pathological changes and the ex-pression of mRNA related to intestinal barrier proteins in the ileum and colon were evaluated using HE stai-ning and RT-qPCR.Results Allicin improved diar-rhea symptoms in mice induced by senna leaf.It re-duced the rate of loose stools,diarrhea index,cumula-tive number of loose stools in five hours,and the intes-tinal propulsion rate.Allicin also protected the intesti-nal mucosa,decreased serum TNF-α and IL-6 levels,and lowered MDA content in the intestines.It in-creased serum NO levels and enhanced SOD and CAT activities in the intestines.Additionally,allicin upreg-ulated the mRNA expression of AQP1,AQP4,and ZO-1 in intestinal tissues.Conclusions Allicin has a significant therapeutic effect on senna-induced diarrhea in mice.The underlying molecular mechanisms may involve anti-inflammatory and antioxidant effects,in-creased NO content,and upregulation of mRNA ex-pression of aquaporins and tight-junction proteins.

BACKGROUND:The AO12-C type middle-proximal humeral fractures are usually caused by high-energy injuries,accompanied by comminuted fractures and a large number of butterfly-shaped bone fragments.These fractures are difficult to achieve good reduction and effective fixation.With the increasing understanding of the biological characteristics of bone and soft tissue,surgical treatment is gradually shifting towards minimally invasive steel plates and intramedullary nail fixation.However,there has been no consensus on which is the best surgical technique for treating humeral fractures in minimal invasive plate osteosynthesis and intramedullary nail.OBJECTIVE:To compare the clinical effect of minimal invasive plate osteosynthesis and intramedullary nail for treating AO12-C type middle-proximal humeral fractures.METHODS:A retrospective analysis was performed in 36 patients with AO12-C type middle-proximal humeral fracture who met the inclusion criteria admitted to the Guizhou Hospital of Beijing Jishuitan Hospital from January 2020 to December 2022.All patients were assigned to minimal invasive plate osteosynthesis group(18 cases)and intramedullary nail group(18 cases)according to the surgical treatment plan.The minimal invasive plate osteosynthesis group received minimally invasive plate osteosynthesis based on preoperative planning assisted by 3D printing,while the intramedullary nail group received intramedullary nail internal fixation.Operation time,intraoperative blood loss,hospital stay,and fracture healing time were compared between the two groups.Visual analog scale score at 1,3,and 6 months after surgery,shoulder joint function,Constant-Murley score,Quick Disabilities of the Arm,Shoulder and Hand(QuickDASH)score,and complications of the shoulder joint at the last follow-up were compared between the two groups.RESULTS AND CONCLUSION:(1)All patients were followed-up for average(15.56±4.05)months,and no difference was observed in hospital stay and fracture healing time between the two groups(P＞0.05).The minimal invasive plate osteosynthesis group had shorter operation time compared to the intramedullary nail group(P＜0.05).The intramedullary nail group had less intraoperative blood loss between the two groups of patients(P＜0.05).(2)In the intramedullary nail group,at 1 and 3 months after operation,the visual analog scale score was significantly lower than the minimal invasive plate osteosynthesis group(P＜0.05).No difference was observed in the visual analog scale in long-term follow-up,shoulder joint function,Constant-Murley score,Quick Disabilities of the Arm,Shoulder and Hand(QuickDASH)score at the last follow-up between the two groups(P＞0.05).(3)No complications such as nonunion or wound infection occurred in either group.Two cases of radial nerve palsy occurred in minimal invasive plate osteosynthesis group,both of which recovered within 3 months.The intramedullary nail group had 1 case of rotator cuff injury.There was no significant difference in the rate of complications between the two groups(P＞0.05).(4)To conclude,minimal invasive plate osteosynthesis and intramedullary nail can achieve good clinical efficacy in the treatment of AO12-C type middle-proximal humeral fractures,effectively improving shoulder joint function in patients.However,minimal invasive plate osteosynthesis exhibits obvious advantages in shorter surgical time with the assistance of 3D printing,which is a valuable,effective,and safe method for treating AO12-C type middle-proximal humeral fractures.

AIM:To establish methods for the isolation,identification,primary culture and evaluation of gas-tric smooth muscle cells(GSMC)from rat and mouse.METHODS:Thirty SD rats and thirty C57BL/6 mice were random-ly assigned into three groups,with 10 rats or mice in each group.Three parallel experiments were conducted to isolate pri-mary GSMC under aseptic conditions.The isolated cells were identified through morphological observation,immunofluo-rescence and Western blot.Flow cytometry was employed to analyze the purity of the cells.Additionally,trypan blue ex-clusion test was utilized to evaluate the viability of the cells after resuscitation.RESULTS:Isolated rat and mouse GSMC exhibited a fusiform or polygonal morphology.Immunofluorescence and Western blot results demonstrated the expression of α-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate of α-SMA was 99.6%in rat GSMC and 97.4%in mouse GSMC.Moreover,the viability rates of rat and mouse GSMC after cryopreservation were found to be greater than 97%.CONCLUSION:A stable and reliable method for the isolation,culture and evaluation of primary GSMC from rat and mouse has been established.

Objective:To explore the spatial correlation between gray matter volume (GMV) changes and neurotransmitter receptors/transporters in patients with first-episode schizophrenia (FES) .Methods:Fifty-four FES patients(FES group) and fifty-nine healthy controls (HC group) were selected from June 2014 to May 2020 in the Psychiatry Department of Renmin Hospital of Wuhan University. Structural magnetic resonance imaging (sMRI) was conducted on all subjects. Differences of GMV were compared across 400 cortical regions and 32 subcortical regions. Based on the positron emission tomography(PET) data from Neuromaps, which provides the density of 19 different neurotransmitter receptors and transporters, Spearman correlation analysis was performed to evaluate the spatial correlation between GMV changes and neurotransmitter systems.Results:Compared to the HC group, FES group exhibited significant GMV reductions in widespread cortical (90/400) and subcortical (6/32) regions (all FDR-corrected P<0.05). The effect size of GMV reduction (Cohen’s d) showed significant positive correlations with the density of 5-hydroxytryptamine 1a(5HT1a) ( r=0.400, Pspin=0.002), γ-aminobutyric acid type A receptor(GABA A)( r=0.307, Pspin=0.002), and metabotropic glutamate receptor 5(mGluR5) ( r=0.275, Pspin=0.020) receptors (all FDR-corrected P<0.05). Conclusion:GMV reductions in a wide range of brain regions existed in patients with FES. There are significant correlations between 5HT1a, GABA A and mGluR5 receptors and gray matter reduction in patients with FES. The disorder of these neurotransmitter receptors may be the potential neurobiological mechanism of gray matter structural abnormalities in the early stage of schizophrenia.

BACKGROUND:The AO12-C type middle-proximal humeral fractures are usually caused by high-energy injuries,accompanied by comminuted fractures and a large number of butterfly-shaped bone fragments.These fractures are difficult to achieve good reduction and effective fixation.With the increasing understanding of the biological characteristics of bone and soft tissue,surgical treatment is gradually shifting towards minimally invasive steel plates and intramedullary nail fixation.However,there has been no consensus on which is the best surgical technique for treating humeral fractures in minimal invasive plate osteosynthesis and intramedullary nail.OBJECTIVE:To compare the clinical effect of minimal invasive plate osteosynthesis and intramedullary nail for treating AO12-C type middle-proximal humeral fractures.METHODS:A retrospective analysis was performed in 36 patients with AO12-C type middle-proximal humeral fracture who met the inclusion criteria admitted to the Guizhou Hospital of Beijing Jishuitan Hospital from January 2020 to December 2022.All patients were assigned to minimal invasive plate osteosynthesis group(18 cases)and intramedullary nail group(18 cases)according to the surgical treatment plan.The minimal invasive plate osteosynthesis group received minimally invasive plate osteosynthesis based on preoperative planning assisted by 3D printing,while the intramedullary nail group received intramedullary nail internal fixation.Operation time,intraoperative blood loss,hospital stay,and fracture healing time were compared between the two groups.Visual analog scale score at 1,3,and 6 months after surgery,shoulder joint function,Constant-Murley score,Quick Disabilities of the Arm,Shoulder and Hand(QuickDASH)score,and complications of the shoulder joint at the last follow-up were compared between the two groups.RESULTS AND CONCLUSION:(1)All patients were followed-up for average(15.56±4.05)months,and no difference was observed in hospital stay and fracture healing time between the two groups(P＞0.05).The minimal invasive plate osteosynthesis group had shorter operation time compared to the intramedullary nail group(P＜0.05).The intramedullary nail group had less intraoperative blood loss between the two groups of patients(P＜0.05).(2)In the intramedullary nail group,at 1 and 3 months after operation,the visual analog scale score was significantly lower than the minimal invasive plate osteosynthesis group(P＜0.05).No difference was observed in the visual analog scale in long-term follow-up,shoulder joint function,Constant-Murley score,Quick Disabilities of the Arm,Shoulder and Hand(QuickDASH)score at the last follow-up between the two groups(P＞0.05).(3)No complications such as nonunion or wound infection occurred in either group.Two cases of radial nerve palsy occurred in minimal invasive plate osteosynthesis group,both of which recovered within 3 months.The intramedullary nail group had 1 case of rotator cuff injury.There was no significant difference in the rate of complications between the two groups(P＞0.05).(4)To conclude,minimal invasive plate osteosynthesis and intramedullary nail can achieve good clinical efficacy in the treatment of AO12-C type middle-proximal humeral fractures,effectively improving shoulder joint function in patients.However,minimal invasive plate osteosynthesis exhibits obvious advantages in shorter surgical time with the assistance of 3D printing,which is a valuable,effective,and safe method for treating AO12-C type middle-proximal humeral fractures.

AIM:To establish methods for the isolation,identification,primary culture and evaluation of gas-tric smooth muscle cells(GSMC)from rat and mouse.METHODS:Thirty SD rats and thirty C57BL/6 mice were random-ly assigned into three groups,with 10 rats or mice in each group.Three parallel experiments were conducted to isolate pri-mary GSMC under aseptic conditions.The isolated cells were identified through morphological observation,immunofluo-rescence and Western blot.Flow cytometry was employed to analyze the purity of the cells.Additionally,trypan blue ex-clusion test was utilized to evaluate the viability of the cells after resuscitation.RESULTS:Isolated rat and mouse GSMC exhibited a fusiform or polygonal morphology.Immunofluorescence and Western blot results demonstrated the expression of α-smooth muscle actin(α-SMA).Flow cytometry showed that the positive expression rate of α-SMA was 99.6%in rat GSMC and 97.4%in mouse GSMC.Moreover,the viability rates of rat and mouse GSMC after cryopreservation were found to be greater than 97%.CONCLUSION:A stable and reliable method for the isolation,culture and evaluation of primary GSMC from rat and mouse has been established.

Aim To study the therapeutic effect of al-licin on senna-induced diarrhea in mice and to explore the underlying mechanism.Methods Forty-eight C57BL/6J mice were randomly divided into six groups:control,model,loperamide positive control group(2 mg·kg-1),allicin low-dose group(6 mg·kg-1),allicin medium-dose group(12 mg·kg-1)and allicin high-dose group(18 mg·kg-1).Except for the con-trol group,the diarrhea model was induced in the other groups by intragastric administration of senna leaf ex-tract.After drug administration,several diarrhea indi-ces were measured:the rate of loose stools,diarrhea index,accumulated frequency of loose stools at differ-ent time points within 5 hours,and small intestine pro-pelling rate.Serum levels of TNF-α and IL-6 were de-tected by ELISA.Serum NO content was determined u-sing the Griess method.The activities of SOD and CAT,as well as MDA content in the ileum and colon,were measured.The pathological changes and the ex-pression of mRNA related to intestinal barrier proteins in the ileum and colon were evaluated using HE stai-ning and RT-qPCR.Results Allicin improved diar-rhea symptoms in mice induced by senna leaf.It re-duced the rate of loose stools,diarrhea index,cumula-tive number of loose stools in five hours,and the intes-tinal propulsion rate.Allicin also protected the intesti-nal mucosa,decreased serum TNF-α and IL-6 levels,and lowered MDA content in the intestines.It in-creased serum NO levels and enhanced SOD and CAT activities in the intestines.Additionally,allicin upreg-ulated the mRNA expression of AQP1,AQP4,and ZO-1 in intestinal tissues.Conclusions Allicin has a significant therapeutic effect on senna-induced diarrhea in mice.The underlying molecular mechanisms may involve anti-inflammatory and antioxidant effects,in-creased NO content,and upregulation of mRNA ex-pression of aquaporins and tight-junction proteins.