[Objective]To summarize Professor LI Xinmin's clinical experience in treating functional constipation in children based on the theory of spleen movement.[Methods]Through learning from the teacher,reading ancient medical books and sorting out clinical medical cases,this paper expounds the theoretical origin of the spleen-transportation theory,analyzes Professor LI's exploration of the etiology and pathogenesis of functional constipation in children from the perspective of"spleen transportation",as well as the clinical experience in treating children's functional constipation with the spleen-transportation theory,and attaches a verified case for evidence.[Results]Professor LI holds that the main etiology of functional constipation in children is the damage to the spleen and stomach caused by dietary stagnation.The key pathogenesis is characterized by"dry fecal accumulation being the secondary aspect with dysfunction of large intestine conduction,and spleen-stomach deficiency being the primary aspect with disharmony of Qi movement in internal organs",among which the disorder of ascending-descending mechanisms in the spleen-stomach and liver-lung axis is particularly prominent.Guided by the"spleen-transportation"theory and the principle of"three-adaptation therapy",clinical treatment employs the strategies of"strengthening the spleen for root cultivation with tonification in transportation,regulating pivotal mechanisms through combined therapies",forming the spleen-transporting pivot-regulation method featured by"dredging-transportation and pivot-regulation".The self-formulated Yunpi Decoction is used to invigorate spleen-earth,focusing on ascending spleen-Yang,regulating middle-Jiao,and regulating Qi movement.In clinical practice,therapies like digestion-promoting,heat-clearing,purging and Qi-supplementing are flexibly integrated based on concomitant syndromes.The harmony method is adeptly applied to reconcile cold-heat and deficiency-excess,emphasizing that medications should be mild to avoid excessive purgation from damaging vital Qi.The pathogenesis of the attached case belonged to spleen deficiency and lack of health,with poor pivot function and internal food stagnation.The treatment adopted the methods of invigorating the spleen and stomach,regulating Qi and promoting smoothness,guiding stagnation and relieving constipation,and administered with the self-designed modified Yunpi Decoction.[Conclusion]Based on the"spleen-transportation theory",Professor LI applies the spleen-transportation and pivot-regulation method featuring dredging-transportation and pivot-regulation.With its unique theory,this approach provides new ideas and methods for clinical practice,making it worthy of promotion and learning.

Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

Phospholipid pharmaceutical excipients are primarily composed of a glycerol backbone,fatty acid chains,and phosphate groups.Due to their amphiphilic structure and unique physicochemical properties,they are commonly used as emulsifiers and carrier materials in pharmaceutical formulations.The pharmacopoeias of various countries have included phospholipid based pharmaceutical excipients and specified the testing methods and limits for their critical quality attributes.This article focuses on the current inclusion status,characteristic indicators,component content determination indicators,and functionality-related indicators determination methods and limits of phospholipid pharmaceutical excipients of various countries,in order to provide references and a basis for the rational applicatione of phospholipid pharmaceutical excipients.

Objective:To evaluate the role of hippocampal activating transcription factor 5 (ATF5) in cognitive impairment induced by neuropathic pain and the relationship with mitochondrial unfolded protein response(mtUPR) in mice.Methods:This study was conducted in 2 parts. Experiment Ⅰ Twenty-four SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups ( n=12 each) using a random number table method: sham operation group (S1 group) and neuropathic pain group (NP group). Neuropathic pain was induced by chronic constriction injury to the sciatic nerve. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before developing the model and at 7, 14, 21 and 28 days after developing the model. Mouse cognitive function was assessed using the novel object recognition test from 30-31 days after developing the model. After the end of the novel object recognition test, mice were sacrificed and the hippocampal CA1 region was harvested for determination of the expression of ATF5 (by Western blot) and the expression of ATF5 in neurons, microglia and astrocytes (by immunofluorescence double staining). Experiment Ⅱ Thirty-six SPF healthy male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=12 each) using a random number table method: sham operation group (S2 group), neuropathic pain + ATF5 up-regulation group (NA group), and neuropathic pain + empty virus group (NE group). On day 14 after developing the model, a virus that specifically up-regulated ATF5 expression in neurons and empty virus were injected into the hippocampal CA1 region. The MWT and TWL were measured at days 28 and 35 after developing the model. The novel object recognition test was performed on day 36 after developing the model to evaluate the cognitive function. After the end of the behavioral test, mice were sacrificed and the hippocampal CA1 region was harvested for detection of the expression of ATF5 and mtUPR marker proteins (Lon protease [LONP1] and heat shock protein 60 [HSP60]) by Western blot. Results:Experiment Ⅰ Compared with S1 group, no statistically significant change was found in the MWT and TWL before developing the model ( P>0.05), the MWT and TWL were significantly decreased on days 7, 14, 21 and 28 after developing the model, the discrimination index (DI) was decreased at day 31 after developing the model, the expression of ATF5 was down-regulated, the expression of ATF5 in neurons was down-regulated ( P<0.05), and no statistically significant change was found in the expression of ATF5 in mircrolia and astrocytes in NP group ( P>0.05). Experiment Ⅱ Compared with S2 group, the MWT and TWL were significantly decreased on days 28 and 35 after developing the model in NE group and NA group, DI was decreased, and the expression of ATF5, LONP1 and HSP60 was down-regulated in NE group ( P<0.05), and no significant change was found in NA group ( P>0.05). Compared with NE group, no significant change was found in the MWT and TWL in NA group ( P>0.05), DI was significantly increased, and the expression of ATF5, LONP1 and HSP60 was up-regulated in NA group ( P<0.05). Conclusions:Down-regulated ATF5 in the hippocampus is involved in the process of cognitive impairment caused by neuropathic pain, and the mechanism may be related to the inhibition of mtUPR.

Phospholipid pharmaceutical excipients are primarily composed of a glycerol backbone,fatty acid chains,and phosphate groups.Due to their amphiphilic structure and unique physicochemical properties,they are commonly used as emulsifiers and carrier materials in pharmaceutical formulations.The pharmacopoeias of various countries have included phospholipid based pharmaceutical excipients and specified the testing methods and limits for their critical quality attributes.This article focuses on the current inclusion status,characteristic indicators,component content determination indicators,and functionality-related indicators determination methods and limits of phospholipid pharmaceutical excipients of various countries,in order to provide references and a basis for the rational applicatione of phospholipid pharmaceutical excipients.

[Objective]To summarize Professor LI Xinmin's clinical experience in treating functional constipation in children based on the theory of spleen movement.[Methods]Through learning from the teacher,reading ancient medical books and sorting out clinical medical cases,this paper expounds the theoretical origin of the spleen-transportation theory,analyzes Professor LI's exploration of the etiology and pathogenesis of functional constipation in children from the perspective of"spleen transportation",as well as the clinical experience in treating children's functional constipation with the spleen-transportation theory,and attaches a verified case for evidence.[Results]Professor LI holds that the main etiology of functional constipation in children is the damage to the spleen and stomach caused by dietary stagnation.The key pathogenesis is characterized by"dry fecal accumulation being the secondary aspect with dysfunction of large intestine conduction,and spleen-stomach deficiency being the primary aspect with disharmony of Qi movement in internal organs",among which the disorder of ascending-descending mechanisms in the spleen-stomach and liver-lung axis is particularly prominent.Guided by the"spleen-transportation"theory and the principle of"three-adaptation therapy",clinical treatment employs the strategies of"strengthening the spleen for root cultivation with tonification in transportation,regulating pivotal mechanisms through combined therapies",forming the spleen-transporting pivot-regulation method featured by"dredging-transportation and pivot-regulation".The self-formulated Yunpi Decoction is used to invigorate spleen-earth,focusing on ascending spleen-Yang,regulating middle-Jiao,and regulating Qi movement.In clinical practice,therapies like digestion-promoting,heat-clearing,purging and Qi-supplementing are flexibly integrated based on concomitant syndromes.The harmony method is adeptly applied to reconcile cold-heat and deficiency-excess,emphasizing that medications should be mild to avoid excessive purgation from damaging vital Qi.The pathogenesis of the attached case belonged to spleen deficiency and lack of health,with poor pivot function and internal food stagnation.The treatment adopted the methods of invigorating the spleen and stomach,regulating Qi and promoting smoothness,guiding stagnation and relieving constipation,and administered with the self-designed modified Yunpi Decoction.[Conclusion]Based on the"spleen-transportation theory",Professor LI applies the spleen-transportation and pivot-regulation method featuring dredging-transportation and pivot-regulation.With its unique theory,this approach provides new ideas and methods for clinical practice,making it worthy of promotion and learning.

Objective To identify and validate co-expressed genes associated with myocardial ischemia/reperfusion injury(MI/RI)and necrotic apoptosis by bioinformatics analysis.Methods Gene expression profile data for MI/RI were obtained by GSE67308 and GSE19875 datasets from the Gene Expression Omnibus(GEO)database.Differential expression analysis was conducted on the GSE67308 dataset to identify differentially expressed genes(DEGs),followed by gene set enrichment analysis and biological pathway analysis.More-over,immune cell infiltration analysis was performed on the GSE67308 dataset.Necrotic apoptosis-related genes were retrieved from the Molecular Signatures Database and the Kyoto Encyclopedia of Genes and Genomes(KEGG).A protein-protein interaction(PPI)network was constructed by overlapping DEGs with these necrotic apoptosis-related genes to identify key genes.Furthermore,the expression pat-terns of these key genes across various cardiac cell types were analyzed using a single-cell sequencing analysis platform,and validation of key gene expression was performed using the GSE19875 dataset.Results A total of 1054 DEGs were identified,comprising 363 upregu-lated and 691 downregulated genes.Gene enrichment analysis revealed that DEGs were primarily associated with processes related to apoptosis,immune responses,and intracellular signaling regulation.Moreover,biological pathway analysis demonstrated that DEGs were predominantly involved in the regulation of signaling pathways such as tumor necrosis factor(TNF)and NF-κB.Immune infiltration anal-ysis indicated a high degree of immune infiltration,particularly with natural killer cells and monocytes,in MI/RI myocardial tissue.PPI network analysis identified Il1b,TNF,Birc3,and Ripk1as crucial genes in the context of necrotic apoptosis.Single-cell sequencing anal-ysis showed the elevated expression of key genes within white blood cells.In comparison to the control group,the MI/RI model group in the GSE19875 dataset exhibited significantly increased expression of Il1b,TNF,Birc3,and Ripk1(P<0.01).Conclusion MI/RI is strongly correlated with the TNF signaling pathway and the NF-κB signaling pathway,both of which play pivotal roles in regulating necrotic apop-tosis.Il1b,TNF,Birc3,and Ripk1emerge as key genes that concurrently regulate both MI/RI and necrotic apoptosis.It is plausible that IL-1b,TNF,Birc3,and Ripk1 may serve as critical regulatory factors in the context of necrotic apoptosis during MI/RI.

Objective:To investigate the incidence and risk factors of coma in patients with hypoglycemia (≤3.9 mmol/L).Methods:A retrospective study was conducted. Patients aged 20 years and older with blood glucose levels ≤3.9 mmol/L, and measured by emergency physicians from January 2020 to December 2022 were collected. Baseline patient data, clinical values collected on-site, and treatment outcomes were analyzed. The Glasgow Coma Scale (GCS) was used to determine if patients were comatose, with GCS ≤8 classified as the coma group and GCS >8 as the non-coma group. Further analysis was conducted on the resuscitated coma group to identify factors affecting patient recovery. Patients were divided into eight age groups, seven time periods within 24 h, and six blood glucose level groups to calculate the incidence of coma. A multivariate logistic regression model was constructed to analyze independent risk factors for coma in hypoglycemic patients.Results:A total of 754 patients with blood glucose levels ≤3.9 mmol/L were collected, with 425 cases of coma and 329 non-coma cases, resulting in a coma probability of 56.37% (95% CI: 52.82%-59.91%). Patients in the coma group were older ( P<0.001) and had a higher prevalence of diabetes compared to the non-coma group (82.12% vs. 67.78%, P<0.001). The age of all patients was (73.05±15.20) years, with the 61-90 years age groups being the most prone to hypoglycemia and coma. In terms of time distribution, the high-incidence periods for hypoglycemia and coma were 0-6 o’clock, 6-9 o’clock, and 14-18 o’clock. The primary causes of hypoglycemia included reduced energy intake after insulin injection (12.07%), improper use of insulin (6.37%), and reduced energy intake (6.23%), with 71.09% of cases having unknown causes. Additionally, 18.44% of patients used insulin before the onset of hypoglycemia, with a higher proportion in the coma group compared to the non-coma group (22.12% vs. 13.68%, P=0.003). The initial blood glucose level of all patients was (2.13±0.85) mmol/L, with lower levels observed in the coma group compared to the non-coma group ( P<0.001). The probabilities of coma occurrence corresponding to blood glucose levels were: 1.1-1.5 mmol/L (72.97%), 1.6-2.0 mmol/L (68.90%), 2.1-2.5 mmol/L (54.10%), 2.6-3.0 mmol/L (38.20%), 3.1-3.5 mmol/L (37.50%), and 3.6-3.9 mmol/L (19.40%). Multivariate logistic regression analysis indicated that age ( OR=1.021, 95% CI: 1.010-1.033, P<0.001), insulin use before onset ( OR=1.948, 95% CI: 1.142-3.323, P=0.014), and blood glucose concentration ( OR=0.426, 95% CI: 0.347-0.522, P<0.001) were independent predictors of coma in hypoglycemic patients. The investigation revealed that after intravenous injection of 50% glucose solution, 215 of 425 coma patients regained consciousness (50.58%), and the recovery time was (18.43±9.09) min. Patients in the recovery group were younger and had lower initial blood glucose levels compared to the non-recovery group (both P<0.05), while recovery group re-measured blood glucose levels were higher than those in the non-recovery group ( P=0.002). Conclusions:The probability of coma in hypoglycemic patients was high, with insulin use being a common trigger. Proper use of insulin is essential to prevent hypoglycemia and coma.

Objective To establish a rapid detection method for invasive candidiasis based on droplet digital polymerase chain reaction(ddPCR).Methods We developed an assay system using a microtitre-based digital PCR platform and designed primer probes specific for four Candida species,namely Candida albicans,Candida smoothii,Candida near-smoothii,and Candida tropicalis.(1)The Limit of Blank(LOB)range and positive judg-ment value were determined by analyzing No Template Control(NTC)samples.(2)The Limit of Detection(LOD)range was determined by diluting positive samples with 10 replicate extractions at each concentration gradient.(3)The Linear Limit of Quantitation(LOQ)range was determined by repetitive testing of diluted samples.(4)The linear range limit was determined through gradient dilution of the positive samples.(5)The coefficient of variation(CV),calculated from the logarithmic values of the resultant concentrations,was assessed by extracting and test-ing positive samples in 12 repetitions at both high and low concentrations.(6)Method reliability was evaluated by calculating the CV from the logarithmic values of the resultant concentrations obtained from clinical samples with fungal culture results.Results The ddPCR assay detected Candida LOB at a range of 0～81 copies/mL,with a positive threshold set at≥3 positive microdroplets.The LOD and LOQ were determined to be 3×102 copies/mL.The linear range for detecting different concentration gradients was found to be between 3×102 and 3×107 copies/mL,with high correlation coefficients observed for Candida albicans(R2=0.999 5),Candida smoothii(R2=0.998 9),Candida near-smoothii(R2=0.999 4),and Candida tropicalis(R2=0.999).Additionally,the coefficient of variation for the resultant concentration logarithmic values was less than 5%,meeting precision requirements.Furthermore,preliminary validation using clinical specimens demonstrated consistent results compared to clinical culture findings.Conclusion ddPCR exhibits rapidity,high sensitivity,good repeatability,and high specificity in detecting inva-sive candidiasis in critically ill patients.This study highlights the potential value of droplet digital PCR as a diag-nostic tool for invasive candidiasis.

[Objective]To introduce and summarize Professor LI Xinmin's experience in treating pediatric epilepsy from the perspective of pivot.[Methods]By following the clinical work,it collected,organized and analyzed initial and follow-up medical records of pediatric epilepsy patients from the First Teaching Hospital of Tianjin University of Traditional Chinese Medicine and consulted ancient Chinese medicine books and modern clinical research on pediatric epilepsy,and analyzed Professor LI's experience in treating pediatric epilepsy from several aspects.Additionally,it attached a medical case for verification.[Results]Professor LI identifies phlegm turbidity and retention as the basic initiating factor of the disease,and phlegm Qi disorder as the core pathogenesis.Combining with the characteristics of children,he utilizes the characteristics of Chinese medicine,using the pivot as the core to treat phlegm and promote Qi flow,supplemented by reconciling the Shaoyang pivot,restoring the circulation of spleen and stomach,and using a dual approach to regulate the overall Qi and coordinate the functions of organs,thereby improving the condition of children with epilepsy and reducing the frequency of seizures.Professor LI also pays attention to individualized treatment,examines the evidence to determine the cause,and adds or subtracts medication according to different types of seizures.At the same time,emphasis is placed on dynamic syndrome differentiation,and medication is prescribed according to the changes in the patient's condition,achieving good therapeutic effects in the treatment and prevention of epilepsy.The attached case was diagnosed as disadvantageous of Shaoyang pivot syndrome at first visit,the treatment was to dredge and benefit Shaoyang,calm the liver and extinguish wind.Chaihu Longgu Muli Decoction and Ditan Decoction were added and subtracted,and then Xieqing Pill was given to clear the liver meridian and remove the heat.[Conclusion]Professor LI's clinical efficacy in treating pediatric epilepsy from the perspective of pivot theory is affirmed,which enriches the diagnosis and treatment system of pediatric epilepsy and is worthy of learning and reference.