ObjectiveTo evaluate the therapeutic effects of Huanglian Jiedutang on cerebral infarction injury in a mouse model of middle cerebral artery occlusion （MCAO） and to explore its mechanism of action on oligodendrocytes， particularly its potential in myelin repair. MethodsMultiple experimental approaches were used to evaluate cerebral ischemic injury and the effects of drug intervention. Laser speckle imaging was used to detect changes in cerebral blood flow， 2，3，5-Triphenyltetrazolium chloride （TTC） staining was used to measure infarct volume， and neurological function was scored according to the Zea-Longa criteria. Brain tissues were routinely embedded in paraffin and subjected to HE and Nissl staining to observe tissue structure and neuronal damage. Animals were divided into a sham group （n=24）， model group （n=24）， Huanglian Jiedutang group （n=24）， and Ginkgo biloba extract （GBE） group （n=18）. After 1 week of acclimatization， intragastric administration was initiated. The sham and model groups received normal saline， the Huanglian Jiedutang group was administered 1.82 g·kg^-1， and the GBE group was administered 0.432 g·kg^-1 after preparation as a 2.16 g·L^-1 solution. All groups were treated for 5 consecutive days at a dose of 0.2 mL·（10 g）^-¹·d^-¹. The MCAO model was established after the final administration on day 6. Single-cell RNA sequencing was used to analyze brain tissue cellular composition and changes in oligodendrocyte subpopulations. Distinct subpopulations were identified by Uniform manifold approximation and projection （UMAP） dimensionality reduction and unsupervised clustering， and marker gene expression was analyzed. Pathway enrichment and causal inference were further performed using IPA. Finally， real-time quantitative PCR was used to verify mRNA expression changes of myelin-related genes. ResultsCompared with the sham group， the model group showed significantly increased neurological function scores （P<0.01）， significantly impaired blood flow （P<0.01）， significantly enlarged cerebral infarct area （P<0.01）， and pathological changes including disordered cortical structural arrangement， aggravated cytoplasmic vacuolization， and increased Nissl bodies. Compared with the model group， the Huanglian Jiedutang and GBE groups showed significantly decreased neurological function scores （P<0.01）， markedly restored blood flow levels （P<0.01）， significantly reduced cerebral infarct area （P<0.01）， and improvement in cortical structural disorder， alleviation of cytoplasmic vacuolization， and a reduction in Nissl bodies. Single-cell data showed that a myelin-associated oligodendrocyte （Mye-OL） subpopulation existed among oligodendrocytes， which was closely related to myelin generation. Compared with the sham group， the number of Mye-OL cells decreased in the model group. Compared with the model group， the number of Mye-OL cells increased in the Huanglian Jiedutang group. This subpopulation promoted the expression of myelin-related genes， including MOG， MBP， and MAG， via transcription factors such as OLIG1， OLIG2， NKX2-2， and SOX10， thereby regulating myelin generation， restoring cognition， and exerting therapeutic effects on acute cerebral infarction. Compared with the sham group， the mRNA expression levels of OLIG1， OLIG2， NKX2-2， and SOX10 were significantly downregulated in the model group （P<0.01）， and the mRNA expression levels of myelin-related genes， including MOG， MBP， and MAG， were also significantly downregulated （P<0.01）. In contrast， compared with the model group， the Huanglian Jiedutang and GBE groups showed significantly upregulated mRNA expression levels of OLIG1， OLIG2， NKX2-2， and SOX10 （P<0.01）， and significantly upregulated mRNA expression levels of myelin-related genes， including MOG， MBP， and MAG （P<0.01）. ConclusionHuanglian Jiedutang exerts therapeutic effects on acute cerebral infarction by regulating the OLIG1/2-NKX2-2-SOX10 signaling pathway to promote myelin generation by Mye-OL cells.

ObjectiveTo investigate the characteristics of metabolic reprogramming during cerebral ischemia-reperfusion injury using single-cell transcriptome sequencing， analyze the heterogeneity of microglial populations， and evaluate the interventional effects of Huanglian Jiedutang on metabolic abnormalities and neuroinflammation. MethodsA transient middle cerebral artery occlusion （tMCAO） model was used to establish ischemic stroke in mice. Local cerebral blood flow changes were monitored by laser speckle imaging. Neurological impairment was evaluated using the Zea-Longa score， and histopathological damage in brain tissue was observed by HE and Nissl staining. Animals were divided into a sham group， model group， Huanglian Jiedutang group， and Ginkgo biloba extract （GBE） group. After 1 week of acclimatization， intragastric administration was initiated. The sham and model groups received normal saline， the Huanglian Jiedutang group was administered 1.82 g·kg^-1， and the GBE group was administered 0.432 g·kg^-1 after preparation as a 2.16 mg/mL solution. All groups were treated for 5 consecutive days （0.2 mL/10 g/day）， and the tMCAO model was established on day 6 after the final administration. At the molecular level， single-cell RNA sequencing was performed on ischemic hemisphere tissue. Non-negative matrix factorization （NMF） was used to cluster microglial subpopulations， combined with differential expression analysis， metabolic reprogramming assessment， and inflammatory factor correlation analysis to elucidate their functional characteristics in ischemia-reperfusion injury. Transcription factor enrichment analysis was further conducted to identify key regulatory nodes. Finally， PCR was used to detect mRNA expression changes of relevant genes to validate the single-cell sequencing results. ResultsCompared with the sham group， the model group showed increased neurological function scores （P<0.01）， decreased blood flow levels （P<0.01）， disordered cortical structure， increased cytoplasmic vacuolization， and increased Nissl bodies. Compared with the model group， the Huanglian Jiedutang and GBE groups showed decreased neurological function scores （P<0.01）， increased blood flow levels （P<0.01）， alleviated cortical structural disorder， reduced cytoplasmic vacuolization， and decreased Nissl bodies. Single-cell analysis showed that microglia could be divided into five subpopulations. Among them， clusters 3 and 5 exhibited significant pro-inflammatory phenotypes， with marked activation of hypoxia and NF-κB signaling pathways， and were identified as pro-inflammatory subpopulations. Clusters 1 and 2 were enriched in Wnt/β-catenin and transforming growth factor（TGF）-β signaling pathways and exhibited prominent anti-inflammatory and reparative characteristics. Meanwhile， glycolysis-related genes， such as HK2， PFKP， and LDHA， were significantly upregulated in the pro-inflammatory subpopulations. Correlation analysis showed that the expression levels of inflammatory molecules were positively correlated with glycolysis-related gene expression levels， whereas the expression levels of reparative and anti-inflammatory molecules were negatively correlated with glycolysis-related gene expression levels， indicating that microglia rely on the glycolytic pathway for energy acquisition under ischemic conditions. Further single-cell transcriptome analysis revealed that Huanglian Jiedutang effectively downregulated key genes driving metabolic reprogramming （such as HK2， PFKP， and LDHA）， significantly reduced the proportion of microglial subpopulations accompanied by glycolytic reprogramming， and inhibited their transformation toward a damage phenotype， thereby reducing inflammatory injury. Meanwhile， compared with the sham group， the mRNA expression levels of interleukin （IL）-1β， IL-6， tumor necrosis factor（TNF）-α， CCL2， CXCL2， and CSF3 were significantly upregulated （P<0.01） in the model group， whereas the mRNA expression levels of endothelial- and pericyte-related functional genes， including RGS5， PECAM1， VEGFB， and NOS3， were significantly downregulated （P<0.01）. In contrast， compared with the model group， the Huanglian Jiedutang and GBE groups showed significantly decreased mRNA expression levels of IL-1β， IL-6， TNF-α， CCL2， CXCL2， and CSF3 （P<0.01）， and significantly increased mRNA expression levels of endothelial- and pericyte-related functional genes， including RGS5， PECAM1， VEGFB， and NOS3 （P<0.01）. ConclusionHuanglian Jiedutang exerts neuroprotective effects by regulating the metabolic reprogramming state of microglia and modulating their inflammatory levels， thereby inhibiting neuroinflammatory injury.

ObjectiveTo evaluate the therapeutic effects of Huanglian Jiedutang on cerebral infarction injury in a mouse model of middle cerebral artery occlusion （MCAO） and to explore its mechanism of action on oligodendrocytes， particularly its potential in myelin repair. MethodsMultiple experimental approaches were used to evaluate cerebral ischemic injury and the effects of drug intervention. Laser speckle imaging was used to detect changes in cerebral blood flow， 2，3，5-Triphenyltetrazolium chloride （TTC） staining was used to measure infarct volume， and neurological function was scored according to the Zea-Longa criteria. Brain tissues were routinely embedded in paraffin and subjected to HE and Nissl staining to observe tissue structure and neuronal damage. Animals were divided into a sham group （n=24）， model group （n=24）， Huanglian Jiedutang group （n=24）， and Ginkgo biloba extract （GBE） group （n=18）. After 1 week of acclimatization， intragastric administration was initiated. The sham and model groups received normal saline， the Huanglian Jiedutang group was administered 1.82 g·kg^-1， and the GBE group was administered 0.432 g·kg^-1 after preparation as a 2.16 g·L^-1 solution. All groups were treated for 5 consecutive days at a dose of 0.2 mL·（10 g）^-¹·d^-¹. The MCAO model was established after the final administration on day 6. Single-cell RNA sequencing was used to analyze brain tissue cellular composition and changes in oligodendrocyte subpopulations. Distinct subpopulations were identified by Uniform manifold approximation and projection （UMAP） dimensionality reduction and unsupervised clustering， and marker gene expression was analyzed. Pathway enrichment and causal inference were further performed using IPA. Finally， real-time quantitative PCR was used to verify mRNA expression changes of myelin-related genes. ResultsCompared with the sham group， the model group showed significantly increased neurological function scores （P<0.01）， significantly impaired blood flow （P<0.01）， significantly enlarged cerebral infarct area （P<0.01）， and pathological changes including disordered cortical structural arrangement， aggravated cytoplasmic vacuolization， and increased Nissl bodies. Compared with the model group， the Huanglian Jiedutang and GBE groups showed significantly decreased neurological function scores （P<0.01）， markedly restored blood flow levels （P<0.01）， significantly reduced cerebral infarct area （P<0.01）， and improvement in cortical structural disorder， alleviation of cytoplasmic vacuolization， and a reduction in Nissl bodies. Single-cell data showed that a myelin-associated oligodendrocyte （Mye-OL） subpopulation existed among oligodendrocytes， which was closely related to myelin generation. Compared with the sham group， the number of Mye-OL cells decreased in the model group. Compared with the model group， the number of Mye-OL cells increased in the Huanglian Jiedutang group. This subpopulation promoted the expression of myelin-related genes， including MOG， MBP， and MAG， via transcription factors such as OLIG1， OLIG2， NKX2-2， and SOX10， thereby regulating myelin generation， restoring cognition， and exerting therapeutic effects on acute cerebral infarction. Compared with the sham group， the mRNA expression levels of OLIG1， OLIG2， NKX2-2， and SOX10 were significantly downregulated in the model group （P<0.01）， and the mRNA expression levels of myelin-related genes， including MOG， MBP， and MAG， were also significantly downregulated （P<0.01）. In contrast， compared with the model group， the Huanglian Jiedutang and GBE groups showed significantly upregulated mRNA expression levels of OLIG1， OLIG2， NKX2-2， and SOX10 （P<0.01）， and significantly upregulated mRNA expression levels of myelin-related genes， including MOG， MBP， and MAG （P<0.01）. ConclusionHuanglian Jiedutang exerts therapeutic effects on acute cerebral infarction by regulating the OLIG1/2-NKX2-2-SOX10 signaling pathway to promote myelin generation by Mye-OL cells.

ObjectiveTo investigate the characteristics of metabolic reprogramming during cerebral ischemia-reperfusion injury using single-cell transcriptome sequencing， analyze the heterogeneity of microglial populations， and evaluate the interventional effects of Huanglian Jiedutang on metabolic abnormalities and neuroinflammation. MethodsA transient middle cerebral artery occlusion （tMCAO） model was used to establish ischemic stroke in mice. Local cerebral blood flow changes were monitored by laser speckle imaging. Neurological impairment was evaluated using the Zea-Longa score， and histopathological damage in brain tissue was observed by HE and Nissl staining. Animals were divided into a sham group， model group， Huanglian Jiedutang group， and Ginkgo biloba extract （GBE） group. After 1 week of acclimatization， intragastric administration was initiated. The sham and model groups received normal saline， the Huanglian Jiedutang group was administered 1.82 g·kg^-1， and the GBE group was administered 0.432 g·kg^-1 after preparation as a 2.16 mg/mL solution. All groups were treated for 5 consecutive days （0.2 mL/10 g/day）， and the tMCAO model was established on day 6 after the final administration. At the molecular level， single-cell RNA sequencing was performed on ischemic hemisphere tissue. Non-negative matrix factorization （NMF） was used to cluster microglial subpopulations， combined with differential expression analysis， metabolic reprogramming assessment， and inflammatory factor correlation analysis to elucidate their functional characteristics in ischemia-reperfusion injury. Transcription factor enrichment analysis was further conducted to identify key regulatory nodes. Finally， PCR was used to detect mRNA expression changes of relevant genes to validate the single-cell sequencing results. ResultsCompared with the sham group， the model group showed increased neurological function scores （P<0.01）， decreased blood flow levels （P<0.01）， disordered cortical structure， increased cytoplasmic vacuolization， and increased Nissl bodies. Compared with the model group， the Huanglian Jiedutang and GBE groups showed decreased neurological function scores （P<0.01）， increased blood flow levels （P<0.01）， alleviated cortical structural disorder， reduced cytoplasmic vacuolization， and decreased Nissl bodies. Single-cell analysis showed that microglia could be divided into five subpopulations. Among them， clusters 3 and 5 exhibited significant pro-inflammatory phenotypes， with marked activation of hypoxia and NF-κB signaling pathways， and were identified as pro-inflammatory subpopulations. Clusters 1 and 2 were enriched in Wnt/β-catenin and transforming growth factor（TGF）-β signaling pathways and exhibited prominent anti-inflammatory and reparative characteristics. Meanwhile， glycolysis-related genes， such as HK2， PFKP， and LDHA， were significantly upregulated in the pro-inflammatory subpopulations. Correlation analysis showed that the expression levels of inflammatory molecules were positively correlated with glycolysis-related gene expression levels， whereas the expression levels of reparative and anti-inflammatory molecules were negatively correlated with glycolysis-related gene expression levels， indicating that microglia rely on the glycolytic pathway for energy acquisition under ischemic conditions. Further single-cell transcriptome analysis revealed that Huanglian Jiedutang effectively downregulated key genes driving metabolic reprogramming （such as HK2， PFKP， and LDHA）， significantly reduced the proportion of microglial subpopulations accompanied by glycolytic reprogramming， and inhibited their transformation toward a damage phenotype， thereby reducing inflammatory injury. Meanwhile， compared with the sham group， the mRNA expression levels of interleukin （IL）-1β， IL-6， tumor necrosis factor（TNF）-α， CCL2， CXCL2， and CSF3 were significantly upregulated （P<0.01） in the model group， whereas the mRNA expression levels of endothelial- and pericyte-related functional genes， including RGS5， PECAM1， VEGFB， and NOS3， were significantly downregulated （P<0.01）. In contrast， compared with the model group， the Huanglian Jiedutang and GBE groups showed significantly decreased mRNA expression levels of IL-1β， IL-6， TNF-α， CCL2， CXCL2， and CSF3 （P<0.01）， and significantly increased mRNA expression levels of endothelial- and pericyte-related functional genes， including RGS5， PECAM1， VEGFB， and NOS3 （P<0.01）. ConclusionHuanglian Jiedutang exerts neuroprotective effects by regulating the metabolic reprogramming state of microglia and modulating their inflammatory levels， thereby inhibiting neuroinflammatory injury.

A total of 44 patients with diabetic foot ulcers were treated in the Traumatic Orthopedics Department of Weifang People′s Hospital from January 2019 to December 2022. After debridement of foot ulcers the wounds were soaked in alkaline water of pH 7.5-8.0 (study group, n=22) or covered with vaseline gauze following iodophor disinfection (control group, n=22). The therapeutic effects of the two methods were compared. Four weeks after debridement, the wound area of study group was smaller than that of control group (3.15 (0, 7.60) vs. 6.75 (3.50, 9.32)cm 2, P<0.05), and the proportion of positive wound bacterial culture was lower than that of control group (40.9% (9/22) vs. 72.7% (16/22), P<0.05). At 12 weeks after surgery, there was no statistically significant difference in the wound healing rate between the two groups (72.7% (16/22) vs. 63.6% (14/22)), but the healing time of the study group was significantly shorter than that of the control group ((6.56±2.68) vs. (9.50±3.87) weeks, P<0.05). It is suggested that immersion of weak alkaline solution is helpful to promote wound healing for patients with diabetic foot ulcers after debridement surgery.

Objective To investigate the effectiveness of esketamine aerosol inhalation combined with Ⅰ-gel laryngeal mask and Wellead bronchial occluder for thoracoscopic single lung ventilation surgery.Methods Seventy-five thoracic surgery patients,39 males and 36 females,aged 28-64 years,BMI 18-25 kg/m2,ASA physical status Ⅰ or Ⅱ,were selected for thoracoscopic surgery.Patients were randomly di-vided into three groups by digital table method:esketamine nebulizing inhalation combined with Ⅰ-gel laryn-geal mask and Wellead bronchial sealer group(group EW),Ⅰ-gel laryngeal mask combined with Wellead bronchial sealer group(group LW),and tracheal catheter combined with Wellead bronchial sealer group(group TW),25 cases in each group.The insertion time,extraction time,awakening time,and the highest PETCO2 value during single lung ventilation were recorded in the three groups.Airway peak pressure(Ppeak)and dynamic lung compliance(Cdyn)were recorded during 15 minutes of supine double-lung ven-tilation and 15 minutes of lateral single-lung ventilation.The changes of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-8(IL-8)and other inflammatory factors in alveolar lavage fluid at 10 mi-nutes before single lung ventilation(T0),10(T1),30(T2),60 minutes(T3)after single lung ventila-tion,and at the end of single lung ventilation(T4),lung collapse score on the operative side,postoperative coughing,hoarseness,sore throat,and atelectasis occurred.Results Compared with group TW,the ex-traction time and awakening time of groups EW and LW were significantly shortened(P＜0.05),Ppeak was significantly decreased and Cdyn was significantly increased in group EW during double lung and single lung ventilation(P＜0.05),and Ppeak was significantly decreased and Cdyn was significantly increased in group LW during single lung ventilation(P＜0.05),TNF-α,IL-6 and IL-8 were significantly decreased at T2-T4 in groups EW and LW(P＜0.05),and the incidence of cough and throat pain was significantly de-creased(P＜0.05).Compared with group LW,Ppeak and Cdyn were significantly decreased in group EW during double lung and single lung ventilation(P＜0.05),and TNF-α,IL-6,and IL-8 were significantly decreased at T2-T4(P＜0.05).There were no significant differences in the insertion time,the highest val-ue of PET CO2 and the lung collapse score among the three groups,and no case of atelectasis occurred among the three groups.Conclusion The use of esketamine aerosol inhalation combined with Ⅰ-gel laryngeal mask and Wellead bronchial occluder for thoracoscopic surgery can ensure the required ventilation during surgery,significantly reduce airway irritation during general anesthesia induction and recovery,reduce pneumonia re-action,and improve lung compliance,which is worthy of clinical promotion.

Objective To observe the long-term effect,adverse reaction and cosmetic outcome of early-stage breast cancer with hypofractionated whole-breast irradiation (HF-WBI) after breast-conserving surgery.Methods A total of 206 patients with stage 0-Ⅱ breast cancer after breast-conserving surgery were included in Shandong Cancer Hospital Affiliated to Shandong University from May 2014 to August 2017.According to radiotherapy fraction,patients were divided into HF-WBI group and conventional whole-breast irradiation (CF-WBI) group.In HF-WBI group,116 patients received whole-breast radiation to 42.56 Gy in 16 fractions followed by tumor bed boost of 9 Gy in 3 fractions or 10 Gy in 5 fractions.In CF-WBI group,90 patients received whole breast radiation to 50 Gy in 25 fractions followed by tumor bed boost of 10 Gy in 5 fractions.The 2-year local recurrence rate,2-year mortality rate,acute adverse reaction,late adverse reaction and cosmetic outcome of the two groups were analyzed.Results The 2-year local recurrence rates of HF-WBI group and CF-WBI group were 0.86％ (1/116) and 2.22％ (2/90) respectively,and there was no significant difference between the two groups (x2 =0.049,P =0.824).The 2-year mortality rates of the two groups were 0.86％ (1/116) and 0 (0/90) respectively,and there was no significant difference (P ＞ 0.999).There were 108 cases (93.1％) in HF-WBI group and 84 cases (93.3％) in CF-WBI group with grade 0-1 acute dermatitis,and 8 cases (6.9％) and 6 cases (6.7％) with grade 2-3 respectively,with no statistically significant difference (x2 =0.004,P =0.948).There were 97 cases (83.6％) in HF-WBI group and 79 cases (87.8％) in CF-WBI group with grade 0-1 bone marrow suppression,and 19 cases (16.4％)and 11 cases (12.2％) with grade 2-4 respectively,with no statistically significant difference (x2 =0.704,P =0.401).In the two groups,there were 1 case (0.9％) and 3 cases (3.3％) with grade 1-2 radiation pneumonitis,and 115 cases (99.1％) and 87 cases (96.7％) with no radiation pneumonitis respectively,and the difference was not statistically significant (x2 =1.626,P =0.202).There was 1 case (0.9％,1.1％) with grade 1 breast edema in each group,and 115 cases (99.1％) and 89 cases (98.9％) did not occur breast edema,with no statistically significant difference (x2 =0.033,P =0.857).In the late adverse reactions,there were 5 cases (4.3％) and 3 cases (3.3％) with skin pigmentation in HF-WBI group and CF-WBI group respectively.There were 2 cases (1.7％,2.2％) with grade 1 subcutaneous tissue fibrosis in each group,and there were 1 case (0.8％) and 2 cases (2.2％) with grade 1 pulmonary fibrosis respectively.The differences between the two groups were not statistically significant (x2 =0.000,P ＞ 0.999;x2 =0.000,P ＞ 0.999;x2 =0.049,P =0.824).The 6-month,1-year and 2-year cosmetic outcome good rates in HF-WBI and CF-WBI group were 96.5％ (111/115) and 93.3％ (84/90),92.1％ (105/114) and 90.0％ (81/90),91.4％ (53/58) and 87.2％ (41/47) respectively.The differences between the two groups were not statistically significant (x2 =0.526,P =0.468;x2 =0.277,P =0.599;x2 =0.476,P =0.490).The whole course of radiotherapy time in HF-WBI group was 25 days or 29 days,which was significantly shorter than the 40 days of CF-WBI group.Conclusion HF-WBI after breast-conserving surgery has the similar long-term effect,acute and late adverse reaction and cosmetic outcome compared with CF-WBI,and the treatment time is significantly shorter.It can be further promoted as the optimal adjuvant radiotherapy for early-stage breast cancer after breast-conserving surgery.

Objective To establish the system of inducement, subculture, and plantlets regeneration of callus from Salvia miltiorrhiza. Methods Compared with the effect of different explants, basic media, and plant growth regulator on the inducement, subculture of callus, and the differentiation of buds. Results MS+6-BA 2 mg/L+NAA 0.2—2 mg/L was propitious to the inducement of callus and the ratio of induced callus, for both the leafstalk and lamina, was 100%, but quantity of induced callus was more of The ratio of buds differentiation was 55.6% on the medium of MS+6-BA 2 mg/L, and it could grown into buds directly. B5+6-BA 1 mg/L+2, 4-D 1 mg/L was the better subculture medium of callus. ConclusionSo for the callus inducement, the better explant and medium are leafstalk and MS+6-BA 2 mg/L+NAA 0.2—2 mg/L, respectively; For the buds differentiation, the better explant and medium are lamina and MS+6-BA 2 mg/L, respectively; For the subculture of callus, B5+6-BA 1 mg/L+2, 4-D 1 mg/L is better.