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 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.

Objective To investigate the effect of pneumoperitoneum pressure during robotic-assisted kidney transplantation (RAKT) on the function of the transplant kidney. Methods The data of 243 kidney transplant recipients were retrospectively analyzed and divided into open kidney transplantation (OKT) group (n=105) and RAKT group (n=138). The RAKT group was further divided into 13 mmHg group (n=67) and 7 mmHg group (n=71) based on pneumoperitoneum pressure. The donor information, recipient's preoperative general data, intraoperative data, and postoperative recovery of the three groups were compared. In the RAKT group, the renal artery, segmental artery, interlobar artery, and venous flow velocity of the transplant kidney were measured using laparoscopic ultrasound. Results There was a statistically significant difference in donor types among the groups (P<0.05), while other donor information and recipient's preoperative general data showed no statistically significant differences (all P>0.05). There were no statistically significant differences in serum creatinine and complications at 30 days and 1 year postoperatively among the groups (all P>0.05). The OKT group and 7 mmHg group had more intraoperative urine output than the 13 mmHg group. Both RAKT groups had less intraoperative blood loss and shorter hospital stays than the OKT group, and longer operation times than the OKT group (all P<0.05). There were no statistically significant differences in operation time, intraoperative blood loss, and hospital stay between the two RAKT groups (all P>0.05). The vascular flow velocity of the transplant kidney decreased at 13 mmHg compared to 7 mmHg pneumoperitoneum pressure, but the differences were not statistically significant (all P>0.05). Conclusions Controllable pneumoperitoneum pressure has a limited impact on the vascular flow velocity of the transplanted kidney. RAKT is a safe and effective surgical method under appropriate pneumoperitoneum pressure, and choosing a lower pneumoperitoneum pressure is more conducive to the early recovery of renal function postoperatively.

Objective To explore the effects of robot assisted training (RAT) combined with neural mobi-lization (NM) training on the recovery of upper limb functions in stroke patients. Methods A total of 110 stroke patients who met the inclusion criteria were selected as the subjects and randomly divided into a control group (n=28),RAT group (n=27),NM group (n=28),and combination group (n=27). All patients underwent routine upper limb occupational therapy. Additionally,the patients in the RAT group were treated with upper limb rehabilitation robots,those in the NM group underwent neural mobilization for treatment,those in the combination group were managed with robot-assisted training for upper limb rehabilitation and neural mobilization. Before treat-ment and 4 weeks after treatment,the modified Ashworth scale (MAS),Fugl-Meyer assessment upper extremity (FMA-UE),functional test for the hemiplegic upper extremity Hong Kong version (FTHUE-HK),and modified Barthel index (MBI) were used to assess the effects. The surface electromyographic signals of the biceps and triceps at the maximum isometric voluntary contraction (MIVC) position during elbow flexion and extension were measured,the integrated electromyographic values (iEMG) were recorded and the synergistic contraction rate (CR) was calculated. Results There was no statistically significant difference (P>0.05) between the four groups in the general information and pre-treatment assessments of MAS,FMA-UE,FTHUE-HK,MBI,iEMG,and CR. After 4 weeks,significant improvements were observed in all indicators compared to the pre-treatment assessments (P<0.05),with the exception of the triceps brachii CR,biceps brachii CR,and elbow extension MIVC biceps brachii iEMG in the control group.Among the group comparisons,all indicators showed statistically significant differences in mean or distribution (P<0.05),except for MAS and triceps brachii CR. The RAT group,NM group,and combination group all demonstrated significant improvements compared to the control group (P<0.05). Nota-bly,the combination group exhibited a greater degree of improvement than the RAT and NM groups. Conclusion RATcombined with NM can reduce upper limb muscle tone in stroke patients. This approacheffectively promotes the establishment of normal movement patterns,improve upper limb motor function,and enhance activities of daily living. This combination is effective and worthy of further clinical promotion and application.

Objective To explore the effects of robot assisted training (RAT) combined with neural mobi-lization (NM) training on the recovery of upper limb functions in stroke patients. Methods A total of 110 stroke patients who met the inclusion criteria were selected as the subjects and randomly divided into a control group (n=28),RAT group (n=27),NM group (n=28),and combination group (n=27). All patients underwent routine upper limb occupational therapy. Additionally,the patients in the RAT group were treated with upper limb rehabilitation robots,those in the NM group underwent neural mobilization for treatment,those in the combination group were managed with robot-assisted training for upper limb rehabilitation and neural mobilization. Before treat-ment and 4 weeks after treatment,the modified Ashworth scale (MAS),Fugl-Meyer assessment upper extremity (FMA-UE),functional test for the hemiplegic upper extremity Hong Kong version (FTHUE-HK),and modified Barthel index (MBI) were used to assess the effects. The surface electromyographic signals of the biceps and triceps at the maximum isometric voluntary contraction (MIVC) position during elbow flexion and extension were measured,the integrated electromyographic values (iEMG) were recorded and the synergistic contraction rate (CR) was calculated. Results There was no statistically significant difference (P>0.05) between the four groups in the general information and pre-treatment assessments of MAS,FMA-UE,FTHUE-HK,MBI,iEMG,and CR. After 4 weeks,significant improvements were observed in all indicators compared to the pre-treatment assessments (P<0.05),with the exception of the triceps brachii CR,biceps brachii CR,and elbow extension MIVC biceps brachii iEMG in the control group.Among the group comparisons,all indicators showed statistically significant differences in mean or distribution (P<0.05),except for MAS and triceps brachii CR. The RAT group,NM group,and combination group all demonstrated significant improvements compared to the control group (P<0.05). Nota-bly,the combination group exhibited a greater degree of improvement than the RAT and NM groups. Conclusion RATcombined with NM can reduce upper limb muscle tone in stroke patients. This approacheffectively promotes the establishment of normal movement patterns,improve upper limb motor function,and enhance activities of daily living. This combination is effective and worthy of further clinical promotion and application.

ObjectiveTo investigate the effect and mechanism of Shenqi Tangluo pill （SQTLP） on oxidative stress injury of skeletal muscle of type 2 diabetes mellitus （T2DM） mice based on nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1）/NAD（P）H quinone oxidoreductase 1 （NQO1） pathway. MethodA total of 60 7-week-old male db/db mice ［specific pathogen-free （SPF） grade］ were selected and fed for one week for adaption. They were divided into the model control group， SQTLP low-， medium- and high-dose （19， 38， and 76 g·kg^-1） groups and metformin group （0.26 g·kg^-1） by gavage. Each group consisted of 12 mice. Twelve male db/m mice of the same age were selected as the blank group. The intervention was implemented continuously for 8 weeks. Fasting blood glucose （FBG） was detected. Fasting serum insulin （FINS） levels were detected by enzyme-linked immunosorbent assay （ELISA）， and the homeostasis model assessment-insulin resistance （HOMA-IR） index and the homeostasis model assessment-insulin sensitivity index （HOMA-ISI） were calculated. Oral glucose tolerance test （OGTT） and insulin tolerance test （ITT） were conducted. The activities of superoxide dismutase （SOD） and glutathione peroxidase （GSH-Px） and the contents of malondialdehyde （MDA） and reduced nicotinamide adenine dinucleotide phosphate （NADPH） in skeletal muscle tissues were detected by biochemical kits. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in skeletal muscle tissues. The levels of reactive oxygen species （ROS） and 4-hydroxynonenal （4-HNE） in skeletal muscle tissue were detected by immunofluorescence （IF）. The expression levels of Nrf2， HO-1， NQO1 and glutamate-cysteine ligase catalytic subunit （GCLC） proteins in skeletal muscle tissues were detected by Western blot. ResultCompared with those in the blank group， FBG， FINS and HOMA-IR in the model group were significantly increased （P<0.05）， while HOMA-ISI was decreased （P<0.05）. The results of OGTT and ITT showed that blood glucose was significantly increased at all time points （P<0.05）， and glucose tolerance and insulin tolerance were significantly impaired. SOD and GSH-Px activities in skeletal muscle tissues were significantly decreased （P<0.05）， and MDA and NADPH contents were significantly increased （P<0.05）. In skeletal muscle tissues， the arrangement of muscle fibers was loose， the nucleus was disordered， and inflammatory cells were infiltrated. The expression levels of ROS and 4-HNE in skeletal muscle tissues were significantly increased （P<0.05）. The protein expression levels of Nrf2， HO-1， NQO1 and GCLC in skeletal muscle tissues were significantly decreased （P<0.05）. Compared with those in the model group， FBG， FINS and HOMA-IR in the metformin group were significantly decreased （P<0.05）， while HOMA-ISI was increased （P<0.05）. The results of OGTT and ITT showed that blood glucose in the metformin group was significantly decreased at all time points （P<0.05）. The activities of SOD and GSH-Px in skeletal muscle tissues were significantly increased （P<0.05）， while the contents of MDA and NADPH were significantly decreased （P<0.05）. No obvious abnormality was found in the skeletal muscle tissue of the metformin group. The expressions of ROS and 4-HNE in skeletal muscle tissues were decreased （P<0.05）. The protein expression levels of Nrf2， HO-1， NQO1 and GCLC in skeletal muscle tissues were significantly increased （P<0.05）. Compared with those in the model group， FBG， FINS and HOMA-IR in the SQTLP medium- and high-dose groups were significantly decreased （P<0.05）， while HOMA-ISI was increased （P<0.05）. The results of OGTT and ITT showed that the glucose tolerance and insulin tolerance of mice were improved in each dose group of SQTLP. The GSH-Px activity in the SQTLP low-dose group was significantly increased （P<0.05）， and the NADPH content was decreased （P<0.05）. The activities of SOD and GSH-Px in the SQTLP medium- and high-dose groups were significantly increased （P<0.05）， while the contents of MDA and NADPH were significantly decreased （P<0.05）. The skeletal muscle tissue injury of mice in each dose group of SQTLP was ameliorated to different degrees. In the SQTLP medium- and high-dose groups， the expressions of ROS and 4-HNE were decreased （P<0.05）， and the protein expression levels of Nrf2， HO-1， NQO1 and GCLC were significantly increased （P<0.05）. Compared with those in the SQTLP low-dose group， FBG and HOMA-IR in the SQTLP high-dose group were significantly decreased （P<0.05）， while HOMA-ISI was increased （P<0.05）. The results of OGTT and ITT showed that the SQTLP high-dose group significantly improved the glucose tolerance and insulin tolerance of mice. The activities of SOD and GSH-Px in skeletal muscle tissues were significantly increased （P<0.05）， while the contents of MDA and NADPH were significantly decreased （P<0.05）. No obvious abnormality was found in the skeletal muscle tissue， the expressions of ROS and 4-HNE were decreased （P<0.05）， and the protein expression levels of Nrf2， HO-1， NQO1 and GCLC were significantly increased （P<0.05） in the skeletal muscle tissue of the SQTLP high-dose group. ConclusionSQTLP can significantly improve IR in T2DM mice， and the mechanism is related to SQTLP activating the Nrf2/HO-1/NQO1 signaling pathway， promoting the expression of antioxidant enzymes， and thus improving the oxidative stress injury in the skeletal muscle.

ObjectiveTo observe the effect of neural mobilization based on shoulder control training on shoulder pain and upper limb function in stroke patients with hemiplegia. MethodsFrom January, 2020 to November, 2021, 43 patients with hemiplegia after stroke in the Second People's Hospital of Nantong were randomly divided into control group (n = 21) and treatment group (n = 22). The control group received shoulder control training, while the treatment group received neural mobilization in addition. Before and after four weeks of treatment, they were evaluated with the Numeric Rating Scale (NRS) of pain and Fugl-Meyer Assessment-Upper Extremities (FMA-UE). ResultsOne case dropped off in the control group and two cases dropped off in the treatment group. After treatment, the NRS score and FMA-UE score improved in both groups (|t| >7.898, P < 0.001), and they were better in the treatment group than in the control group (|t| >2.337, P < 0.05). ConclusionNeural mobilization based on shoulder control training can significantly alleviate shoulder pain and improve upper limb motor function in stroke patients with hemiplegia.

Objective To explore the learning curve of robotic-assisted kidney transplantation(RAKT).Methods The clinical data of 96 consecutive RAKT patients performed by the same surgical team were retrospectively analyzed.The arterial anastomosis time,venous anastomosis time,ureteral anastomosis time,hospital stay,and blood loss were selected as evaluation indicators.The learning curve of RAKT was analyzed using the cumulative sum(CUSUM),and the curve was divided into the learning improvement stage and the proficient mastery stage according to the learning curve.The learning curve was verified by comparing the general data and surgical data of patients in different learning stages,and the clinical efficacy of each stage was analyzed.Results The optimal fitting equation of the learning curve reached its peak at the 33rd case,which was the minimum number of surgeries required to master RAKT.There was no statistically significant difference in age,gender,dialysis type,previous abdominal surgery history,number of donor renal arteries,and preoperative serum creatinine between the learning improvement group and the proficient mastery group(all P>0.05).Compared with the learning improvement stage,the body mass index(BMI)was higher,and the number of right donor kidney was increased compared to the left donor kidney in the proficient mastery stage(both P<0.05).There were no significant differences in arterial anastomosis time,ureteral anastomosis time,postoperative serum creatinine,and complications between the two groups(all P>0.05).The iliac vessel dissection time,warm ischemia time,venous anastomosis time,blood loss,and hospital stay in the proficient mastery stage were superior to those in the learning improvement stage,with statistically significant differences(all P<0.05).Conclusions RAKT requires at least 33 cases to cross the learning curve.There is no difference in complications and recovery of transplant renal function between the learning improvement stage and the proficient mastery stage.

OBJECTIVE To establish high performance liquid chromatography(HPLC)fingerprints of the extract of Fructus Li-gustri Lucidi-Ecliptae Herba and to identify the structure of its chemical constituents.METHODS The chromatographic separation was performed on ZORBAX Extend C18(250 mm×4.6 mm,5 μm)column with the mobile phase consisting of acetonitrile and 0.1%formic acid solution in gradient elution mode at a flow rate of 1.0 mL·min-1.The optimum column temperature was set at 35℃,the detection wavelength was 265 nm,and the injection volume was 10 μL.The HPLC fingerprints of 15 batches of herb pair from different origins were established.Then,similarity evaluation,cluster analysis,and orthogonal partial least squares discriminant analysis were carried out.The chemical structures of the constituents of the herb pair were identified based on the high-resolution mass spectrometry utilizing the UPLC-Q-Exactive Orbitrap MS in positive and negative-ion modes.RESULTS As a result,the similarity of the 15 bat-ches of samples varied from 0.865 to 0.992.A total of 16 common peaks were identified in the fingerprints,and 7 of them were identi-fied by comparision with the reference substances.Furthermore,55 chemical compounds were detected,and 7 of them were accurately identified based on the reference substances,and the chemical structures of the rest 48 components were temporarily speculated.CON-CLUSION The fingerprint method established in this study is simple,reliable,and reproducible,and can be further used for the quality control of Fructus Ligustri Lucidi-Ecliptae Herba herb pair.The developed UPLC-Q-Exactive Orbitrap MS method can be applied to identify the structures of chemical constituents in this herb pair and lay a foundation for further research.

Glycolysis is an important biological event in the metabolic reprogramming process of primary liver cancer.Its is mainly regulated by key rate-limiting enzymes in the glycolysis pathway,including hexokinase(HK),pyruvate kinase(PK),phosphofructokinase(PFK),and lactate dehydrogenase(LDH).Moreover,it can also be regulated by multiple mechanisms such as glucose transporters(GLUTS),monocarboxylic acid transporters(MCT),PI3K/AKT/mTORC signaling pathway and hypoxia induction factor(HIF).More and more studies have proved that key glycolytic enzymes and regulatory factors play important roles in hepatocellular carcinoma(HCC)cell proliferation,invasion,metastasis,immune escape,and drug resistance.Currently,with the continuous in-depth research on the mechanism of glycolysis,clinical therapies targeting glycolysis has become a new therapeutic strategy for HCC treatment.This article aims to summarize the research progress of key glycolytic enzymes and regulatory factors in the occurrence and development of primary liver cancer,hoping to provide help for the prevention and treatment of liver cancer.