ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design（QbD） and to assess its health risk. MethodsTaking whole iron content， Fe²⁺ dissolution content and looseness as critical quality attributes（CQAs）， and calcination temperature， calcination time， spreading thickness and particle size as critical process parameters（CPPs） determined by the failure mode and effect analysis（FMEA）， the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation（AHP-CRITIC） hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry， and the health risk assessment was carried out by daily exposure（EXP）， target hazard quotient（THQ） and lifetime cancer risk（LCR）， and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃， calcination time of 1 h， particle size of 0.2-0.5 cm， spreading thickness of 1 cm， and vinegar quenching for 1 time［Haematitum-vinegar（10：3）］. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1， while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10^-6-1×10^-4， and the LCR of the rest was<1×10^-6， and the LCRs of calcined Haematitum were all<1×10^-6， indicating that the carcinogenic risk of calcined Haematitum was low， but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg were formulated as 1 014， 25， 17， 27， 7 mg·kg^-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible， and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.

ObjectiveTo investigate the processing technology of calcined Haematitum based on the concept of quality by design（QbD） and to assess its health risk. MethodsTaking whole iron content， Fe²⁺ dissolution content and looseness as critical quality attributes（CQAs）， and calcination temperature， calcination time， spreading thickness and particle size as critical process parameters（CPPs） determined by the failure mode and effect analysis（FMEA）， the processing technology of calcined Haematitum was optimized by orthogonal test combined with analytic hierarchy process-criteria importance through intercriteria correlation（AHP-CRITIC） hybrid weighting method. The contents of heavy metals and harmful elements were determined by inductively coupled plasma mass spectrometry， and the health risk assessment was carried out by daily exposure（EXP）， target hazard quotient（THQ） and lifetime cancer risk（LCR）， and the theoretical value of the maximum limit was deduced. ResultsThe optimal processing technology for calcined Haematitum was calcination at 650 ℃， calcination time of 1 h， particle size of 0.2-0.5 cm， spreading thickness of 1 cm， and vinegar quenching for 1 time［Haematitum-vinegar（10：3）］. The contents of 5 heavy metals and harmful elements in 13 batches of calcined Haematitum were all decreased with reductions of up to 5-fold. The cumulative THQ of 2 batches of samples was>1， while the cumulative THQ of all batches of Haematitum was>1. The LCR of As in 1 batches of Haematitum was 1×10^-6-1×10^-4， and the LCR of the rest was<1×10^-6， and the LCRs of calcined Haematitum were all<1×10^-6， indicating that the carcinogenic risk of calcined Haematitum was low， but special attention should still be paid to Haematitum medicinal materials. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg were formulated as 1 014， 25， 17， 27， 7 mg·kg^-1. ConclusionThe optimized processing technology of calcined Haematitum is stable and feasible， and the contents of heavy metals and harmful elements are reduced after processing. Preliminary theoretical values of the maximum limits of Cu， As， Cd， Pb and Hg are formulated to provide a scientific basis for the formulation of standards for the limits of harmful elements in Haematitum.

ObjectiveTo explore the mechanism of Huanglian Wendantang on damp-heat type diabetes enteropathy rats based on the G protein coupled bile acid receptor 5/glucagon like peptide-1 （TGR5/GLP-1） signaling pathway and intestinal flora. MethodsA total of 72 male Sprague Dawley （SD） rats were adaptively fed for one week. Twelve SD rats were randomly selected as a blank group and fed with an ordinary diet. The rest of the SD rats were fasted for 12 hours without water. A rat model with damp-heat type diabetes enteropathy was made by left intraperitoneal injection of streptozotocin （55 mg·kg^-1） and high sugar and high fat diet （20% sucrose solution + high fat diet） in a humid and hot environment （artificial climate box： temperature 30-34 ℃， relative humidity： 85%-95%）. After successful modeling， the rats were randomly divided into a model group， a metformin group （200 mg·kg^-1）， low-dose， medium-dose， and high-dose Huanglian Wendantang groups （7.10， 14.20， 28.39 g·kg^-1）， with 12 rats in each group. The normal group and the model group were orally administered with physiological saline once a day for 6 consecutive weeks. During the observation period， the weight and blood glucose levels of rats were measured and recorded weekly. After the administration， fresh feces were collected from rats， and 16S rRNA sequencing technology was used to study the differences and changes in intestinal flora among different groups. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the serum of rats were detected by enzyme-linked immunosorbent assay （ELISA）， and the pathological morphological changes of colon tissue were examined. The expression of TGR5 and GLP-1 in colon tissue was detected by immunohistochemistry， and the expression of TGR5 and GLP-1 proteins in colon tissue was measured by Western blot. ResultsCompared with the blank group， the model group showed a decrease in body weight， an increase in blood glucose， and significant damp-heat symptoms. The levels of IL-6 and TNF-α in serum were significantly increased （P<0.01）. The expression of TGR5 and GLP-1 was decreased （P<0.01）， and the pathogenic bacteria were increased. Compared with the model group， the treatment groups exhibited improvements in body weight， blood glucose levels， and damp-heat syndrome in rats. Among them， the high-dose group of Huanglian Wendantang displayed the most significant improvement effect， with significantly reduced inflammation levels （P<0.01） and elevated expression of TGR5 and GLP-1 （P<0.01）. Colonic pathological sections showed that Huanglian Wendantang could effectively ameliorate colonic pathological changes. The 16S rRNA sequencing result indicated a significant increase in beneficial bacteria in the treatment groups. ConclusionHuanglian Wendantang can effectively ameliorate the damp-heat symptoms and blood glucose levels in rats with damp-heat type diabetes enteropathy， and it may exert an effect by regulating the TGR5/GLP-1 signaling pathway and intestinal flora disorder.

Objective:To explore the expression of connexin 43 (Cx43) in hippocampus of post-stroke depression (PSD) model rats and its effect on cell apoptosis and depressive-like behavior.Methods:Sixty SPF-grade male SD rats aged 6-8 weeks were randomly divided into five groups (12 rats in each group): normal group, stroke group, depression group, PSD group and carbenoxolone(CBX) group. The stroke model was established by injection of endothelin-1.Chronic unpredictable mild stress (CUMS) combined with solitary rearing was used to establish a depression model. Rats in PSD group were given CUMS and raised alone on the seventh day of stroke modeling.Rats in CBX group were given intraperitoneal injection of CBX(20 mg/kg) on 14th day after PSD modeling. The depressive-like behavior of rats was evaluated by sugar water preference test and open field test. The expression of Cx43 mRNA in hippocampus of rats was detected by RT-PCR, the expression levels of Cx43, caspase-3, Bax and Bcl-2 were detected by Western blot, and the changes of apoptosis rate were detected by TUNEL staining. SPSS 23.0 software was used for statistical analysis, the behavioral data were analyzed by repeated measurement ANOVA, the remaining data were analyzed by one-way ANOVA, and the LSD- t test was used for further pairwise comparison. Results:(1)As for the preference rate of sugar water and the times of crossing the grid, the interaction effects between time and group were significant among the 5 groups( Finteraction=35.57, 111.43, both P<0.05). On the 28th day after operation, the preference rate of sugar water and the times of crossing grid in depression group and PSD group were lower than those in stroke group (all P<0.05), while the preference rate of sugar water and the times of crossing grid in CBX group were both lower than those in PSD group (both P<0.05). (2) The levels of Cx43 mRNA and Cx43 protein in the five groups were significantly different ( F=273.57, 64.56, both P<0.05). The levels of Cx43 mRNA and Cx43 protein in depression group ((0.59±0.05), (0.69±0.08)) and PSD group ((0.61±0.07), (0.63±0.12)) were lower than those in stroke group ((1.01±0.03), (1.05±0.08)) (all P<0.05). The levels of Cx43 mRNA and Cx43 protein in CBX group ((0.30±0.01), (0.37±0.09)) were lower than those in PSD group (both P<0.05). (3) The protein levels of caspase-3, Bax, Bcl-2 and Bcl-2/Bax and the apoptosis rate of the five groups were significantly different ( F=102.40, 90.27, 47.42, 159.99, 115.21, all P<0.05). The levels of caspase-3, Bax protein, apoptosis rate in stroke group ((0.44±0.06), (0.54±0.07), (29.16±5.03)) and depression group ((0.45±0.07), (0.59±0.09), (27.00±4.93)) were higher than those in normal group ((0.21±0.08), (0.33±0.07), (4.83±3.18)) (all P<0.05), the levels of Bcl-2 protein and Bcl-2/Bax in stroke group ((0.80±0.04), (1.51±0.20)) and depression group ((0.60±0.09), (1.03±0.09)) were lower than those in normal group ((1.04±0.13), (3.14±0.38)) (all P<0.05).The levels of caspase-3, Bax protein and apoptosis rate in PSD group ((0.76±0.05), (0.84±0.02), (44.50±3.83)) were all higher than those in stroke group and depression group (all P<0.05), and the levels of Bcl-2 protein and Bcl-2/Bax in PSD group ((0.50±0.14), (0.59±0.17)) were lower than those in stroke group and depression group (both P<0.05). The levels of caspase-3 and Bax protein and the apoptosis rate in CBX group ((1.03±0.10), (1.02±0.05), (56.00±4.81)) were higher than those in PSD group (all P<0.05).The levels of Bcl-2 protein and Bcl-2/Bax in CBX group((0.26±0.08), (0.25±0.08)) were lower than those in PSD group (both P<0.05). Conclusion:The expression level of Cx43 in the hippocampus of PSD model rats is downregulated, which can promote cell apoptosis and exacerbate depressive behavior.

Objective:To evaluate the role of hippocampal prostaglandin-endoperoxide synthase 2 (PTGS2) in baicalin-induced reduction of cognitive dysfunction after cerebral ischemia-reperfusion injury (CIRI) in mice.Methods:Thirty healthy male C57BL/6 mice, aged 16 weeks, weighing 20-25 g, were divided into 5 groups ( n=6 each) using a random number table method: control group (C group), CIRI group, baicalin+ CIRI group (B+ CIRI group), overexpression of PTGS2+ CIRI group (PTGS2+ CIRI group), and overexpression of PTGS2+ baicalin+ CIRI group (PTGS2+ B+ CIRI). In B+ CIRI group and PTGS2+ B+ CIRI group, baicalin-liposome 0.2 ml was injected through the tail vein, and the CIRI model was established 1 week later. In PTGS2+ CIRI group and PTGS2+ B+ CIRI group, PTGS2-overexpressed adeno-associated virus 1.2 μl was injected into the hippocampus, and the CIRI model was established 4 weeks later. CIRI model was established by using the transient (50 min) bilateral common carotid artery occlusion/reperfusion. On the 12th day after developing the model, the spatial learning and memory ability was evaluated using Morris water maze test. The expression of PTGS2 in the hippocampus was detected by Western blot, and the expression of tumor necrosis factor-alpha (TNF-α), interleukin-1beta (IL-1β), IL-6, Iba-1 and CD68 mRNA in the hippocampus was detected by quantitative real-time polymerase chain reaction. Results:Compared with C group, the escape latency was significantly prolonged, the time spent in the target quadrant was shortened, the number of crossing the original platform was reduced, and the expression of PTGS2 and expression of Iba-1, CD68, TNF-α, IL-1β and IL-6 mRNA in the hippocampus was up-regulated in CIRI group ( P<0.05). Compared with CIRI group, the escape latency was significantly shortened, the time spent in the target quadrant was prolonged, the number of crossing the original platform was increased, and the expression of PTGS2 and expression of Iba-1, CD68, TNF-α, IL-1β and IL-6 mRNA in the hippocampus was down-regulated in B+ CIRI group, and the escape latency was significantly prolonged, the time spent in the target quadrant was shortened, the number of crossing the original platform was reduced, and the expression of PTGS2 and expression of Iba-1, CD68, TNF-α, IL-1β and IL-6 mRNA in the hippocampus was up-regulated in PTGS2+ CIRI group ( P<0.05). Compared with B+ CIRI group, the escape latency was significantly prolonged, the time spent in the target quadrant was shortened, the number of crossing the original platform was reduced, and the expression of PTGS2 and expression of Iba-1, CD68, TNF-α, IL-1β and IL-6 mRNA in the hippocampus was up-regulated in PTGS2+ B+ CIRI group ( P<0.05). Conclusions:The mechanism by which baicalin attenuates cognitive dysfunction after CIRI is related to down-regulation of hippocampal PTGS2 expression and inhibition of neuroinflammation in mice.

Objective:To observe the changes of peripheral blood T cell subsets and serum tumor necrosis factor (TNF)-α and interleukin (IL)-6 contents in narcolepsy type 1 (NT1) patients and their correlations with narcolepsy, and provide basis for finding the biological markers of narcolepsy.Methods:A retrospective analysis was performed. From March 2022 to December 2023, 23 patients with NT1 admitted to Epilepsy and Sleep Disorder Center, Second Affiliated Hospital of Harbin Medical University and 23 healthy controls underwent physical examination of nervous system in our center were enrolled. T lymphocyte subsets CD4 + and CD8 + in peripheral blood were calculated by flow cytometry. Serum TNF-α and IL-6 contents were detected by enzyme-linked immunosorbent assay. Multivariate Logistic regression was used to determine the correlations of NT1 with CD4 + T lymphocyte count and IL-6 and TNF-α contents, and diagnostic values of CD4 + T lymphocyte and TNF-α in NT1 were evaluated via area under receiver operating characteristics (ROC) curve. Results:Compared with the healthy controls, the NT1 patients had significantly increased peripheral blood CD4 + T lymphocyte count ([820.61±316.87] /μL vs. [1121.04±387.47] /μL), and significantly higher serum TNF-α and IL-6 contents ([39.97±10.64] pg/mL vs. [57.01±19.92] pg/mL; [22.50±6.09] pg/mL vs. [33.66±17.28] pg/mL, P<0.05). No significant difference in peripheral blood CD8 + T lymphocyte count was noted between the 2 groups ([668.65±276.45] pg/mL vs. [592.52±217.78] pg/mL, P>0.05). Multivariate Logistic regression showed that CD4 + T lymphocyte count and serum TNF-α content were independent risk factors for NT1 ( OR=1.004, 95% CI: 1.001-1.006, P=0.007; OR=1.133, 95% CI: 1.032-1.243, P=0.009). Area under ROC curve of the two combined indexes was 0.881(95% CI: 0.784-0.977, P=0.001), enjoying sensitivity of 0.783 and specificity of 0.870. Conclusion:Combination of peripheral blood CD4 + T lymphocyte count and serum TNF-α content has high diagnostic performance in predicting NT1.

Continuous inflammatory response and repeated tissue damage lead to a chronic inflammatory state that is an important pathological feature of chronic refractory wounds, and plays an important pathogenic role in the occurrence and development of chronic refractory wounds. Multiple types of programmed cell death can release damage-associated molecular patterns, and trigger or induce sustained inflammatory responses, leading to dysregulated tissue repair. This review will outline the known role of programmed cell death in the inflammatory response of chronic refractory wounds from the perspective of generating damage-associated molecular patterns, and explore possible research directions on the role and mechanism of programmed cell death in chronic refractory wounds.

Objective:To prepare hydroxyapatite(HA)coating on polyether ether ketone(PEEK)surface by magnetron sputtering technique and to study its biosafety.Methods:Sulfonated PEEK was used to increase the binding area and HA coating was constructed on it using magnetron sputtering technology.SEM and energy dispersive spectroscopy(EDAX)were used to detect the construction effect.Cell adhesion assay,cytoskeletal fluorescence staining and SEM validation were used to assess cytologrcal safety.In vivo safety tests were conducted in SD rats and golden hamsters.Results:HA coating with gradient morphology was successfully constructed on the PEEK surface using above technique.The coating promoted cell adhesion,extension and proliferation.No systemic toxicity and no sig-nificant influence in HE staining of the main infernal organs samples were observed.The coating alleviated the oral mucosal irritation caused by simple sulfonation to a certain extent.Conclusion:HA coating can be prepared stably with magnetron sputtering technology and can meet the biosafety needs for clinical applications.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Liver regeneration following injury aids the restoration of liver mass and the recovery of liver function. In the present study we investigated the contribution of megakaryocytic leukemia 1 (MKL1), a transcriptional modulator, to liver regeneration. We report that both MKL1 expression and its nuclear translocation correlated with hepatocyte proliferation in cell and animal models of liver regeneration and in liver failure patients. Mice with MKL1 deletion exhibited defective regenerative response in the liver. Transcriptomic analysis revealed that MKL1 interacted with E2F1 to program pro-regenerative transcription. MAPKAPK2 mediated phosphorylation primed MKL1 for its interaction with E2F1. Of interest, phospholipase d2 promoted MKL1 nuclear accumulation and liver regeneration by catalyzing production of phosphatidic acid (PA). PA administration stimulated hepatocyte proliferation and enhanced survival in a MKL1-dependent manner in a pre-clinical model of liver failure. Finally, PA levels was detected to be positively correlated with expression of pro-regenerative genes and inversely correlated with liver injury in liver failure patients. In conclusion, our data reveal a novel mechanism whereby MKL1 contributes to liver regeneration. Screening for small-molecule compounds boosting MKL1 activity may be considered as a reasonable approach to treat acute liver failure.