ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley （SD） rats were randomly divided into a normal group， model group （M）， Sini San group （MS， 2.5 g·kg^-1）， and fluoxetine group （MF， 2 mg·kg^-1）. Except for the normal group， rats in the other three groups were subjected to chronic unpredictable mild stress （CUMS）. After 8 weeks， the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum corticosterone （CORT）， adrenocorticotropic hormone （ACTH）， corticotropin-releasing factor （CRF）， lipopolysaccharide （LPS）， Zonulin， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， γ-aminobutyric acid （GABA） levels in the hippocampus and prefrontal cortex， and brain-derived neurotrophic factor （BDNF） levels in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy， and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group， the sucrose preference of rats in the model group was significantly reduced （P0.01）， whereas it was significantly increased in the Sini San group compared with the model group （P0.05）. Compared with the normal group， hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased （P0.05）， and compared with the model group， both were significantly increased in the Sini San group （P0.05， P0.01）. Compared with the normal group， serum LPS and Zonulin levels in the model group were significantly increased （P0.05， P0.01）， and compared with the model group， Zonulin levels in the Sini San group were significantly decreased （P0.05）. No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group， widened and blurred tight junctions， sparse and shortened microvilli， and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group， which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators， liver function， and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.

ObjectiveTo explore the efficacy and mechanisms of Sini San in the treatment of depression and liver injury based on gut microbiota. MethodsThirty-two male Sprague-Dawley （SD） rats were randomly divided into a normal group， model group （M）， Sini San group （MS， 2.5 g·kg^-1）， and fluoxetine group （MF， 2 mg·kg^-1）. Except for the normal group， rats in the other three groups were subjected to chronic unpredictable mild stress （CUMS）. After 8 weeks， the open-field test and sucrose preference test were conducted. Enzyme-linked immunosorbent assay （ELISA） was used to detect serum corticosterone （CORT）， adrenocorticotropic hormone （ACTH）， corticotropin-releasing factor （CRF）， lipopolysaccharide （LPS）， Zonulin， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， γ-aminobutyric acid （GABA） levels in the hippocampus and prefrontal cortex， and brain-derived neurotrophic factor （BDNF） levels in the hippocampus. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect hippocampal BDNF mRNA expression. Serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） levels were measured using the ultraviolet lactate dehydrogenase method. The ultrastructure of the intestinal epithelium was observed by electron microscopy， and gut microbiota in rat feces were analyzed using 16S rDNA high-throughput sequencing. ResultsCompared with the normal group， the sucrose preference of rats in the model group was significantly reduced （P<0.01）， whereas it was significantly increased in the Sini San group compared with the model group （P<0.05）. Compared with the normal group， hippocampal GABA protein levels and BDNF mRNA expression in the model group were significantly decreased （P<0.05）， and compared with the model group， both were significantly increased in the Sini San group （P<0.05， P<0.01）. Compared with the normal group， serum LPS and Zonulin levels in the model group were significantly increased （P<0.05， P<0.01）， and compared with the model group， Zonulin levels in the Sini San group were significantly decreased （P<0.05）. No obvious changes were observed in the ultrastructure of the jejunal mucosa among groups. Compared with the normal group， widened and blurred tight junctions， sparse and shortened microvilli， and mitochondrial swelling with cristae disruption in epithelial cells were observed in the ileal and colonic mucosa of the model group， which were markedly improved in the Sini San and fluoxetine groups. The results of 16S rDNA high-throughput sequencing showed that Sini San improved CUMS-induced dysbiosis of Bacteroidetes and Proteobacteria. Correlation analysis indicated that Bacteroidetes and Proteobacteria were significantly correlated with depression-related indicators， liver function， and intestinal mucosal permeability. ConclusionSini San exerts antidepressant and hepatoprotective effects by improving Bacteroidetes and Proteobacteria and inhibiting the increase in intestinal mucosal permeability in CUMS rats.

ObjectiveBased on the concept of quality by design（QbD）， the processing process of calcined Pyritum was optimized， and its X-ray diffraction（XRD） fingerprint was established. MethodsThe safety， effectiveness and quality controllability of calcined Pyritum were taken as the quality profile（QTPP）， the color， hardness， metallic luster， phase composition， the contents of heavy metals and hazardous elements were taken as the critical quality attributes（CQAs）， and the calcination temperature， calcination time， paving thickness and particle size were determined as the critical process parameters（CPPs）. Differential thermal analysis， X-ray diffraction（XRD） and inductively coupled plasma mass spectrometry（ICP-MS） were used to analyze the correlation between the calcination temperature and CQAs of calcined Pyritum. Then， based on the criteria importance through intercriteria correlation（CRITIC）-entropy weight method， the optimal processing process of calcined Pyritum was optimized by orthogonal test. Powder XRD was used to analyze the phase of calcined Pyritum samples processed according to the best process， and the mean and median maps of calcined Pyritum were established by the superposition of geometric topological figures， and similarity evaluation and cluster analysis were carried out. ResultsThe results of single factor experiments showed that the physical phase of Pyritum changed from FeS₂ to Fe₇S₈ during the process of temperature increase， the color gradually deepened from dark yellow， and the contents of heavy metals and harmful elements decreased. The optimized processing process of calcined Pyritum was as follows：calcination temperature at 750 ℃， calcination time of 2.5 h， paving thickness of 3 cm， particle size of 0.8-1.2 cm， vinegar quenching 1 time［Pyritum-vinegar（10∶3）］. After calcination， the internal structure of Pyritum was honeycomb-shaped， which was conducive to the dissolution of active ingredients. XRD fingerprints of 13 batches of calcined Pyritum characterized by 10 common peaks were established. The similarities of the relative peak intensities of the XRD fingerprints of the analyzed samples were>0.96， and it could effectively distinguish the raw products and unqualified products. ConclusionTemperature is the main factor affecting the quality of calcined Pyritum. After processing， the dissolution of the effective components in Pyritum increases， and the contents of heavy metals and harmful substances decrease， reflecting the function of processing to increase efficiency and reduce toxicity. The optimized processing process is stable and feasible， and the established XRD fingerprint can be used as one of the quality control standards of calcined Pyritum.

ObjectiveBased on the concept of quality by design（QbD）， the processing process of calcined Pyritum was optimized， and its X-ray diffraction（XRD） fingerprint was established. MethodsThe safety， effectiveness and quality controllability of calcined Pyritum were taken as the quality profile（QTPP）， the color， hardness， metallic luster， phase composition， the contents of heavy metals and hazardous elements were taken as the critical quality attributes（CQAs）， and the calcination temperature， calcination time， paving thickness and particle size were determined as the critical process parameters（CPPs）. Differential thermal analysis， X-ray diffraction（XRD） and inductively coupled plasma mass spectrometry（ICP-MS） were used to analyze the correlation between the calcination temperature and CQAs of calcined Pyritum. Then， based on the criteria importance through intercriteria correlation（CRITIC）-entropy weight method， the optimal processing process of calcined Pyritum was optimized by orthogonal test. Powder XRD was used to analyze the phase of calcined Pyritum samples processed according to the best process， and the mean and median maps of calcined Pyritum were established by the superposition of geometric topological figures， and similarity evaluation and cluster analysis were carried out. ResultsThe results of single factor experiments showed that the physical phase of Pyritum changed from FeS₂ to Fe₇S₈ during the process of temperature increase， the color gradually deepened from dark yellow， and the contents of heavy metals and harmful elements decreased. The optimized processing process of calcined Pyritum was as follows：calcination temperature at 750 ℃， calcination time of 2.5 h， paving thickness of 3 cm， particle size of 0.8-1.2 cm， vinegar quenching 1 time［Pyritum-vinegar（10∶3）］. After calcination， the internal structure of Pyritum was honeycomb-shaped， which was conducive to the dissolution of active ingredients. XRD fingerprints of 13 batches of calcined Pyritum characterized by 10 common peaks were established. The similarities of the relative peak intensities of the XRD fingerprints of the analyzed samples were>0.96， and it could effectively distinguish the raw products and unqualified products. ConclusionTemperature is the main factor affecting the quality of calcined Pyritum. After processing， the dissolution of the effective components in Pyritum increases， and the contents of heavy metals and harmful substances decrease， reflecting the function of processing to increase efficiency and reduce toxicity. The optimized processing process is stable and feasible， and the established XRD fingerprint can be used as one of the quality control standards of calcined Pyritum.

Objective To investigate the characteristics of epilepsy onset based on the theory of five circuits and six qi in traditional Chinese medicine(TCM).Methods A total of 8 389 epilepsy patients from a community-based natural population cohort study from UK Biobank were included.Frequency and constituent ratio analyses were performed to describe the distribution of epilepsy onset in relation to the five circuits and six qi elements,and statistical inferences were made using chi-square tests.Results The analysis revealed statistically significant differences existed in epilepsy onset across different heavenly stems,earthly branches,recombinant yearly circuit,celestial control and terrestrial effect,and dominant qi(all P＜0.05).From the perspective of circuits,the highest incidence occurred in years corresponding to the"Ji"and"Hai"years,while the lowest incidence was observed in"Bing"and"Mao"years.Years dominated by wood circuit and wind manifestations exhibited the highest incidence,whereas years with cold manifestations showed the lowest incidence,suggesting the liver as the primary pathological site in epilepsy.From the perspective of qi,the third dominant qi was most frequently associated with onset.Regarding presiding and subordinate qi,years with taiyin damp earth presiding and taiyang cold water subordinate had the highest incidence,while years with yangming dry metal presiding and shaoyang monarch fire subordinate had the lowest incidence,indicating that cold-damp years and midsummer periods are potential triggers for epilepsy.Conclusion There is a certain regularity in the epilepsy onset correlated with the features of five circuits and six qi,which can guide prevention and clinical diagnosis and treatment of epilepsy.

Objective:To investigate the effects and mechanisms of angiotensin-converting enzyme 2 (ACE2) and Captopril (CAP) on mechanical ventilation-induced lung injury (VILI) in mice.Methods:Seventy-two healthy male BALB/c mice were randomly assigned (using a random number table) into six groups ( n=12 per group): normal control (NC) group, VILI group, ACE2 group, VILI+ACE2 group, CAP group, and VILI+CAP group. One hour prior to mechanical ventilation, the ACE2 and VILI+ACE2 groups were intraperitoneally injected with ACE2 at a dose of 0.1 mg/kg, while the CAP and VILI+CAP groups were intraperitoneally injected with CAP at a dose of 2.5 mg/kg. Following mechanical ventilation, serum samples were collected and enzyme-linked immunosorbent assay (ELISA) was used to detect inflammatory factors [platelet activating factor (PAF), endothelin-1 (ET-1), soluble intercellular adhesion molecule-1 (sICAM-1), prostaglandin E2 (PGE2)] and cardiovascular system related indicators [von Willebrand factor (vWF), thrombomodulin (TM), angiotensin (Ang) (1-9), Ang (1-7), prostacyclin I 2 (PGI2)]. Bronchoalveolar lavage fluid (BALF) was gathered, and total protein concentration was determined using BCA method, and sICAM-1 levels were measured by ELISA. Lung tissues were collected and subjected to hematoxylin and eosin staining (HE staining) for the assessment of pathological lung injury and lung injury scoring. Western blot and reverse transcription quantitative polymerase chain reaction (RT-qPCR) were utilized to detect the relative expression levels of ACE2 protein and mRNA, respectively. Statistical analysis was performed using IBM SPSS 20.0 software. Intergroup comparisons were conducted using one-way analysis of variance followed by the least significant difference (LSD) test. Results:No statistically significant differences were observed in the levels of PAF, ET-1, sICAM-1, vWF, TM, Ang(1-9), Ang(1-7), and PGI2 in serum and lung tissues between the ACE2/CAP groups and the NC group (all P>0.05). Compared with the VILI group, the VILI+ACE2 and VILI+CAP groups exhibited significantly decreased serum and lung tissue levels of PAF, ET-1, sICAM-1, and vWF (all P<0.05), while the levels of TM, Ang(1-9), Ang(1-7), and PGI2 were significantly increased (all P<0.05). Pathological lung injury was alleviated, and the lung wet/dry weight ratio was significantly reduced (all P<0.05) in the VILI+ACE2 and VILI+CAP groups. Furthermore, both ACE2 protein and mRNA expression levels were significantly increased in these groups (all P<0.05). Conclusion:Both ACE2 and CAP can inhibit inflammation and protect the cardiovascular system, possibly by promoting the ACE2/Ang(1-9)/Ang(1-7) axis, thereby exerting a protective effect against VILI.

Objective:To develop a prediction model for the dosage of antivenom in patients envenomed by Trimeresurus stejnegeri (bamboo pit viper) using Lasso-Logistic regression.Methods:A retrospective cohort study was conducted including 165 patients with Trimeresurus stejnegeri bites admitted to the Emergency General Ward of Yangjiang Hospital Affiliated to Guangdong Medical University (Yangjiang People's Hospital) between October 31, 2021, and November 1, 2024. Data on demographic characteristics, key clinical signs, laboratory parameters, antivenom treatment regimen, and total antivenom dosage were collected. Predictive variables were selected using LASSO and Logistic regression methods. The prediction model was constructed using R software. Model calibration was assessed using the Hosmer-Lemeshow test, discriminative ability via the receiver operating characteristic (ROC) curve, and clinical utility using decision curve analysis.Results:Among the 165 enrolled patients, 71 received a total antivenom dose of ≤3 vials, and 94 received >3 vials. Six predictors for requiring >3 vials of antivenom were identified: pain severity, swelling degree, serum treatment regimen, 3P-test result, white blood cell count (WBC), and fibrinogen level (FIB). After comparing 22 prediction models incorporating 4 to 6 variables, the model comprising swelling degree, pain severity, WBC, and FIB was selected. The calibration curve indicated good agreement between predicted and actual risks. The area under the ROC curve (AUC) was 0.78 (95% CI: 0.71–0.85), demonstrating satisfactory discriminative ability for predicting whether the total antivenom dose exceeded 3 vials. Conclusions:The risk prediction model shows good performance in identifying whether the total antivenom dose exceeds 3 vials in Trimeresurus stejnegeri bite patients. It incorporates six readily available clinical variables: pain severity, swelling degree, serum regimen, 3P-test, WBC, and FIB. A model based on pain severity, swelling degree, WBC, and FIB can assist clinicians in predicting whether a patient will require more than 3 vials of antivenom after standard treatment.

Objective To observe the effects and mechanism of Guiqi Yiyuan Ointment on immune function in 12C6+ beam bystander-effect rats.Methods Totally 60 Wistar rats were randomly divided into blank group,model group,positive control group,and TCM low-,medium-,and high-dosage groups,with 10 rats in each group.TCM low-,medium-,and high-dosage groups were given 1.2,2.4,and 4.8 g/kg Guiqi Yiyuan Ointment for gavage,and the blank group,model group and positive control group were given equal dose of sodium chloride solution for gavage for 14 days.Positive control group received intraperitoneal injection of amifostine 30 minutes before modeling.On the 15th day,except the blank group,the other groups were given 4 Gy 12C6+ beam of single irradiation on the right lung.After irradiation,each group was given gavage for 3 days.The general condition of rats was observed,peripheral blood leukocyte(WBC),red blood cell(RBC),hemoglobin(Hb),platelet(PLT)levels were detected,ELISA was used to detect the contents of serum interleukin(IL)-4 and interferon-γ(IFN-γ),lung and spleen organ indices were calculated,HE staining was used to observe the pathological morphology of left lung and spleen tissues,and flow cytometry was used to detect the proportion of CD4+ and CD8+ T lymphocytes in the spleen,qPCR was used to detect the expressions of T-bet and GATA-3 mRNA in right lung and spleen tissues.Results Compared with the blank group,the model group rats showed a decrease in peripheral blood WBC,an increase in RBC,Hb,PLT,and an decrease in serum IL-4 and IFN-γ contents;the organ indices of left lung and spleen was significantly reduced,the alveoli were deformed,the alveolar walls became thicker,the alveolar cavities shrank,the lung tissue was significantly bleeding,the spleen tissue structure was severely damaged,the splenic sinus cavity became narrower,and the splenic cord showed fibrous hyperplasia;the proportion of CD4+ T lymphocytes and CD4+/CD8+ ratio in spleen tissue of the model group decreased,while the proportion of CD8+ T lymphocytes increased,the expression of T-bet mRNA in the left lung and spleen tissues decreased,while the expression of GATA-3 mRNA increased,with statistical significance(P<0.05).Compared with the model group,the positive control group and TCM medium-and high-dosage groups showed an increase in peripheral blood WBC,an decrease in RBC,Hb,PLT,an increase in serum IL-4 and IFN-γ contents;the index of left lung and spleen organs increased in TCM high-dosage group;the pathological damage of left lung and spleen tissues in each treatment group alleviated to varying degrees,while the proportion of CD4+ T lymphocytes and CD4+/CD8+ ratio in spleen tissue of the positive control group and TCM high-dosage group increased,the proportion of CD8+ T lymphocytes decreased,the expression of T-bet mRNA in left lung and spleen tissues increased,and the expression of GATA-3 mRNA decreased,with statistical significance(P<0.05).Conclusion Guiqi Yiyuan Ointment can alleviate the lung and spleen injury of rats induced by 12C6+ beam radiation.Its mechanism is related to regulating the proportion of CD4+,CD8+ T lymphocytes and the expression of T-bet and GATA-3,and recovering cellular immune function.

In order to realize the diagnosis of slit lamp in cross-regional patients and improve the real-time and convenience of diagnosis,a remote slit lamp diagnosis platform based on Internet of Things(IoT)technology is designed.Firstly,the feasibility of remote slit lamp is analyzed.Secondly,the IoT platform architecture of doctor/server/facility(D/S/F)is proposed and a remote slit lamp is designed.Finally,the performance of the remote slit lamp diagnostic platform is tested.The platform solves the communication problem of distributed slit lamps and realizes respectively numerical control of multi-area slit lamp by multi-eye experts.The test results show that the remote control delay of the platform is less than 20 ms,which supports multiple experts to diagnose multiple patients separately.

Objective:To explore the in vitro antibacterial effects of low-intensity pulsed ultrasound (LIPUS) combined with 3.5 g/L povidone iodine solution on the biofilm of methicillin-resistant staphylococcus aureus (MRSA). Methods:Immature (cultured for 24 hours) and mature (cultured for 72 hours) MRSA biofilms were established on the surfaces of glass slides or confocal dishes. They were randomly divided into 4 groups ( n=9) according to different intervention methods. In the control group, glass slides or confocal dishes were placed in 500 mL of physiological saline for 3 minutes; in the PI group, glass slides or confocal dishes were placed in 500 mL of 3.5 g/L povidone iodine solution for 3 minutes; in the LIPUS group, glass slides or confocal dishes were placed in 500 mL of physiological saline and simultaneously intervened with LIPUS for 3 minutes; in the LIPUS & PI group, glass slides or confocal dishes were placed into 500 mL of 3.5 g/L povidone iodine solution and simultaneously intervened with LIPUS for 3 minutes. After intervention, confocal microscopy (CLSM) and scanning electron microscopy (SEM) were used to observe and compare the structure, morphology, bacterial survival, and viable cell count of the MRSA biofilms among the 4 groups. Results:On the MRSA biofilms cultured for 24 and 72 hours, CLSM and SEM observed sparse biofilms in the LIPUS group and LIPUS & PI group, and also a large number of dead bacteria in the LIPUS & PI group. On the MRSA biofilms cultured for 24 hours, the bacterial colony counts in the control group, PI group, LIPUS group, and LIPUS & PI group were (1.21±0.45)×10 6 CFU/mL, (3.38±2.81)×10 3 CFU/mL, (1.82±0.37)×10 3 CFU/mL, and (69.67±27.93) CFU/mL, respectively. Except for the comparison between PI group and LIPUS group, which showed no statistically significant difference ( P>0.05), there were statistically significant differences between the other groups when compared pairwise ( P<0.05). On the MRSA biofilms cultured for 72 hours, the bacterial colony counts in the control group, PI group, LIPUS group, and LIPUS & PI group were (3.01±0.70)×10 6 CFU/mL, (1.80±1.52)×10 5 CFU/mL, (2.10±0.52)×10 3 CFU/mL, and (68.67±19.55) CFU/mL, respectively. There were statistically significant differences between the 4 groups when compared pairwise ( P<0.05). Conclusions:Application of LIPUS or 3.5 g/L povidone iodine alone for 3 minutes on the immature or mature MRSA biofilms in vitro only leads to partial antibacterial activity. However, LIPUS can enhance the in vitro antibacterial effect of 3.5 g/L povidone iodine on the MRSA biofilms at different maturity levels.