Type 2 diabetes mellitus （T2DM） and hypertension are common and frequent chronic non-communicable diseases， which often coexist in clinical practice， resulting in a large number of cardiovascular events and deaths， and their case fatality rate far exceeds that of other factors such as dyslipidemia and obesity. Based on the diagnostic standards， guidelines， and animal model evaluation methods of T2DM with hypertension at home and abroad， this study summarized， evaluated， and analyzed the characteristics of the existing animal models of T2DM with hypertension based on the clinical symptoms in traditional Chinese and Western medicine. The animal models showing high fitting degrees with the clinical symptoms in Western medicine are mainly established by injection of streptozocin （STZ） in SHR rats in the surgical induction and chemical induction methods and feeding a high-fat and high-salt diet combined with STZ injection in SD rats in the dietary induction methods. The models showcasing high fitting degrees with the clinical symptoms in traditional Chinese medicine （TCM） are mainly established by the surgical induction method. Considering the fitting degrees and the advantages and disadvantages， the ideal modeling method for T2DM with hypertension is the two-kidney， one clip （2K1C） method （a surgical induction method） combined with feeding a high-fat and high-sugar diet and STZ injection. However， the available models lack the characteristics of TCM and the evaluation indicators have poor specificity. This study found that there are few animal models of T2DM with hypertension considering the characteristics of both disease and syndrome， which may be related to the identification and attribution of TCM syndromes in animal macroscopic information. In view of this problem， it is suggested that the evaluation criteria should be established and improved for the animal models combining disease and syndrome， which can help to evaluate the fitting degree of the pathological characteristics of different syndromes in the animal models of T2DM with hypertension. In this way， ideal animal models of T2DM with hypertension can be established to simulate the disease occurrence and development in the human body. The animal models are expected to provide an ideal approach for the further research on the pathogenesis of T2DM and its prevention and treatment with TCM， which is of great significance for the treatment and prevention of T2DM with hypertension and the prognosis of its complications. At the same time， breakthroughs in the basic syndrome models of comorbidities are expected to lay a foundation for the leapfrog development of TCM research.

Type 2 diabetes mellitus （T2DM） and hypertension are common and frequent chronic non-communicable diseases， which often coexist in clinical practice， resulting in a large number of cardiovascular events and deaths， and their case fatality rate far exceeds that of other factors such as dyslipidemia and obesity. Based on the diagnostic standards， guidelines， and animal model evaluation methods of T2DM with hypertension at home and abroad， this study summarized， evaluated， and analyzed the characteristics of the existing animal models of T2DM with hypertension based on the clinical symptoms in traditional Chinese and Western medicine. The animal models showing high fitting degrees with the clinical symptoms in Western medicine are mainly established by injection of streptozocin （STZ） in SHR rats in the surgical induction and chemical induction methods and feeding a high-fat and high-salt diet combined with STZ injection in SD rats in the dietary induction methods. The models showcasing high fitting degrees with the clinical symptoms in traditional Chinese medicine （TCM） are mainly established by the surgical induction method. Considering the fitting degrees and the advantages and disadvantages， the ideal modeling method for T2DM with hypertension is the two-kidney， one clip （2K1C） method （a surgical induction method） combined with feeding a high-fat and high-sugar diet and STZ injection. However， the available models lack the characteristics of TCM and the evaluation indicators have poor specificity. This study found that there are few animal models of T2DM with hypertension considering the characteristics of both disease and syndrome， which may be related to the identification and attribution of TCM syndromes in animal macroscopic information. In view of this problem， it is suggested that the evaluation criteria should be established and improved for the animal models combining disease and syndrome， which can help to evaluate the fitting degree of the pathological characteristics of different syndromes in the animal models of T2DM with hypertension. In this way， ideal animal models of T2DM with hypertension can be established to simulate the disease occurrence and development in the human body. The animal models are expected to provide an ideal approach for the further research on the pathogenesis of T2DM and its prevention and treatment with TCM， which is of great significance for the treatment and prevention of T2DM with hypertension and the prognosis of its complications. At the same time， breakthroughs in the basic syndrome models of comorbidities are expected to lay a foundation for the leapfrog development of TCM research.

OBJECTIVE: To propose a machine learning-based method for predicting the trajectories during manual acupuncture manipulation (MAM), aiming to improve the precision and consistency of acupuncture practitioner' operation and provide the real-time suggestions on MAM error correction. METHODS: Computer vision technology was used to analyze the hand micromotion when holding needle during acupuncture, and provide a three-dimensional coordinate description method of the index finger joints of the holding hand. Focusing on the 4 typical motions of MAM, a machine learning-based MAM trajectory predictive model was designed. By integrating the changes of phalangeal joint angle and hand skeletal information of acupuncture practitioner, the motion trajectory of the index finger joint was predicted accurately. Besides, the roles of machine learning-based MAM trajectory predictive model in the skill transmission of acupuncture manipulation were verified by stratified randomized controlled trial. RESULTS: The performance of MAM trajectory predictive model, based on the long short-term memory network (LSTM), obtained the highest stability and precision, up to 98%. The learning effect was improved when the model applied to the skill transmission of acupuncture manipulation. CONCLUSION The machine learning-based MAM predictive model provides acupuncture practitioner with precise action prediction and feedback. It is valuable and significant for the inheritance and error correction of manual operation of acupuncture.
Humans ; Acupuncture Therapy/instrumentation* ; Machine Learning ; Adult ; Male ; Female

In protein engineering, while computational models are increasingly used to predict mutation effects, their evaluations primarily rely on high-throughput deep mutational scanning (DMS) experiments that use surrogate readouts, which may not adequately capture the complex biochemical properties of interest. Many proteins and their functions cannot be assessed through high-throughput methods due to technical limitations or the nature of the desired properties, and this is particularly true for the real industrial application scenario. Therefore, the desired testing datasets, will be small-size (∼10-100) experimental data for each protein, and involve as many proteins as possible and as many properties as possible, which is, however, lacking. Here, we present VenusMutHub, a comprehensive benchmark study using 905 small-scale experimental datasets curated from published literature and public databases, spanning 527 proteins across diverse functional properties including stability, activity, binding affinity, and selectivity. These datasets feature direct biochemical measurements rather than surrogate readouts, providing a more rigorous assessment of model performance in predicting mutations that affect specific molecular functions. We evaluate 23 computational models across various methodological paradigms, such as sequence-based, structure-informed and evolutionary approaches. This benchmark provides practical guidance for selecting appropriate prediction methods in protein engineering applications where accurate prediction of specific functional properties is crucial.

ObjectiveTo investigate the effect and possible mechanism of Shenfu Injection （参附注射液） on rat cardiomyocyte injury induced by isoproterenol from the perspective of regulating the high mobility group protein B1 （HMGB1）/ Toll-like receptor 4 （TLR4）/nuclear factor κB （NF-κB） pathway through the deacetylation modification of silent information regulator 1 （SIRT1）. MethodsThe optimal concentration and intervention duration of isoproterenol hydrochloride and the optimal intervention concentration of Shenfu Injection were screened out by CCK-8 method. Logarithmically growing H9c2 rat cardiomyocytes were taken at 5×10⁴ cells/well and divided into normal group， model group， Shenfu Injection group， and SIRT1 inhibitor group， with 3 replicates in each group.Except for the normal group， the cells in the other groups were induced by isoproterenol hydrochloride to establish a chronic heart failure cell model. After modeling， the Shenfu Injection group was given Shenfu Injection at the optimal intervention concentration， and the SIRT1 inhibitor group was given 1 μmol/L of SIRT1 inhibitor EX-527， for optimal intervention duration.CCK-8 assay was used to detect the cell activity and calculate the inhibitory rate. ELISA assay was used to detect the nicotinamide adenine dinucleotide oxidation state/ nicotinamide adenine dinucleotide reduction state （NAD+/NADH） in cardiomyocytes. Immunofluorescence was used to detect the immunofluorescence localization of HMGB1 and SIRT1 in cardiomyocytes. Western blotting was used to detect the protein expression of acetylated HMGB1 in cardiomyocytes， HMGB1 in the nucleus and cytoplasm， and SIRT1， TLR4， myeloid differentiation factor 88 （MYD88） and NF-κB p65 in cardiomyocytes. RT-qPCR was used to detect the mRNA expression of SIRT1， HMGB1， TLR4， MYD88 and NF-κB p65 in cardiomyocytes. ResultsThe optimal intervention concentration of isoproterenol hydrochloride was 300 μmol/L， and the intervention duration was 48 hours； 8% was the optimal intervention concentration of Shenfu Injection. Compared to those in the normal group， the cell activity， NAD+/NADH value， nuclear HMGB1 protein expression， cardiomyocyte SIRT1 protein and mRNA expression in the model group decreased， while the cell inhibition rate， cardiomyocyte acetylated HMGB1 and cytoplasmic HMGB1 protein expression， cardiomyocyte TLR4， MYD88， NF-κB p65 protein and mRNA expression all increased （P＜0.05）； fluorescence localization showed that the content of HMGB1 in cardiomyocytes in the model group increased and was localized in both the nucleus and cytoplasm.Compared to the model group and the SIRT1 inhibitor group， the Shenfu Injection group showed significant improvements in all the above indicators （P＜0.05）； fluorescence localization showed that the SIRT1 content increased in the Shenfu injection group， while the HMGB1 content decreased， and was mainly located in the nucleus. ConclusionShenfu Injection can improve myocardial cell damage by increasing SIRT1 expression to reduce the acetylation level of HMGB1， regulating the HMGB1/TLR4/NF-κB pathway and inhibiting the nuclear translocation of HMGB1.

ObjectiveTo evaluate the clinical effectiveness and safety of Lianling Formula （连苓方） in the treatment of adolescent atopic dermatitis （AD） exacerbation stage with heart fire and spleen deficiency syndrome. MethodsNinety-two patients were randomly divided into a treatment group and a control group， with 46 cases in each group. Patients in the control group took oral ebastine tablets， and patients in the treatment group took Lianling Formula orally， and both groups used 0.03% topical tacrolimus for 4 weeks. Before and after treatment， the improvement of scoring atopic dermatitis index by 50% （SCORAD50） and 75% （SCORAD75）， scoring atopic dermatitis index （SCORAD）， visual analog scale （VAS）， traditional Chinese medicine （TCM） symptom score， the children's dermatology life quality index （CDLQI）， Athens insomnia scale （AIS） score of both groups were evaluated， and the adverse reactions in both groups were recorded. ResultsFour cases in the treatment group and five cases in the control group were dropped， and intention-to-treat analysis was used. Finally 46 cases in each group were included in the statistics. The SCORAD scores after treatment were lower than those before treatment， and the scores in treatment group were lower than those of the control group （P＜0.01）； the attainment rates of SCORAD50 and SCORAD75 after treatment in the treatment group were 80.43% and 47.83%， which were significantly higher than those in the control group of 63.04% and 26.09%， respectively （P＜0.05）. VAS scores， TCM symptom scores， CDLQI scores， and AIS scores were lower than those before treatment （P＜0.01）， and VAS scores， TCM symptom scores， and CDLQI scores of the treatment group were lower than those of the control group after treatment （P＜0.05 or P＜0.01）， while AIS scores were higher than those of the control group （P＜0.05）. The erythema， papule/edema， oozing/crusting， epidermal peeling， and dryness in the SCORAD scores of the treatment group after treatment were lower than those before treatment （P＜0.05 or P＜0.01）， and the erythema， papule/edema， oozing/crusting， and dryness of children in the control group were lower than those before treatment （P＜0.05 or P＜0.01）. The scores of erythema， oozing/crusting， and epidermal peeling in the treatment group after treatment were all lower than those in the control group （P＜0.05）. The scores of vexation， itching， dry mouth， poor appetite and irregular bowel movements in terms of TCM symptoms in the treatment group after treatment were lower than those before treatment （P＜0.01）， and the scores of itching， vexation， dark urine in the control group were lower than those before treatment （P＜0.05 or P＜0.01）. The scores of itching， dry mouth， poor appetite， irregular bowel movements， and dark urine in the treatment group after treatment were all lower than that in the control group （P＜0.05 or P＜0.01）. No adverse reactions occurred in both groups of children during treatment. ConclusionLianling Formula combined with topical tacrolimus can effectively improve the clinical symptoms of adolescent AD exacerbation with heart fire and spleen deficiency syndrome， and the formula is effective and safe in improving the children's skin lesions， reducing itching， relieving the accompanied TCM symptoms， and improving the quality of life.

Objective To construct methoxy polyethylene glycol (mPEG) modified gold nanoparticles (AuNPs) loaded with doxorubicin (DOX) AuNPs-mPEG@DOX in order to reduce the toxicity and side effects of DOX. Methods AuNPs-mPEG@DOX was prepared and characterized by Z-Average, Zeta potential and UV-Vis spectroscopy. The impact of thiol-linked DOX (HS-DOX) at various dosage concentrations on the drug adsorption rate and drug loading of AuNPs-mPEG@DOX was investigated. Furthermore, a HPLC method was developed to accurately determine the content of unadsorbed HS-DOX in AuNPs-mPEG@DOX. The specificity, linearity, precision, stability and average recovery of this method were thoroughly investigated. The cytotoxic effect of AuNPs-mPEG@DOX on MCF-10A and MCF-7 cells was evaluated using a CCK-8 assay. Results AuNPs-mPEG@DOX was successfully prepared with Z-Average of (46.12±0.49) nm, Zeta potential of (18.60±1.51) nm and the maximum absorption wavelength of 530 nm. An efficient HPLC method for the detection of unadsorbed HS-DOX in AuNPs-mPEG@DOX was devised. The optimal dosage concentration of HS-DOX for AuNPs-mPEG@DOX was determined to be 11.18 μg/ml, resulting in a drug adsorption rate of (9.21±2.88)% and a drug loading rate of (2.01±0.62)%. Cytotoxicity experiments demonstrated that AuNPs-mPEG@DOX significantly reduced the toxic and side effects of DOX on normal breast cells. Additionally, AuNPs-mPEG@DOX and free DOX exhibited comparable cytotoxic effects on breast tumor cells when DOX concentration was equal to or greater than 4.75 μmol/L. Conclusion AuNPs-mPEG@DOX effectively reduce the toxicity of DOX, providing a reference for future research on reducing the toxicity of AuNPs-linked drugs.

Objective:To detect itching,anxiety,depression behaviors in chronic itch models of mice and observe the activation of γ-aminobutiric acid(GABA)neurons in the ventral sector of the zona incerta(ZIv),and provide mor-phological evidence for their involvement in the modulation of itch information.Methods:Diphenylcyclopropenone(DCP)was used in glutamic acid decarboxylase 67-green fluorescent protein(GAD67-GFP)knock-in mice to establish chronic itch model.Itch behaviors were detected by video tracking system to verify whether the models were successfully established.The anxiety,depression behaviors of chronic itch model mice were detected by using elevated plus maze test(EPM)and tail suspention test(TST).By using GAD67-GFP mice,the distribution of GABAergic neurons in va-rious sectors of the zona incerta(ZI)was observed.And combined with immunofluorescence staining method,double labeling of GABAergic neurons with FOS in ZIv were observed respectively in control and DCP group mice.Results:In brain slices of GAD67-GFP mice,GABAergic neurons can be observed within all sectors of ZI and are more concentrat-ed in ZIv.Compared with control group mice,DCP group mice showed a significant increase in the bouts of scratching(P＜0.001).The time of immobility in TST was significantly higher in DCP group mice than in control group mice,which displayed depression-like behavior.The EPM test showed that the numbers of entries and proportion of time in the cross region in DCP group mice were less than in control group mice.EPM test revealed that DCP group mice exhibited anxiety-like behavior.The results of immunofluorescence staining showed that the number of FOS-positive cells in ZIv was significantly higher in DCP group mice than in control group mice,and abundant co-labeled neurons of FOS and GABAergic neurons were observed in ZIv.Conclusion:GABAergic neurons were predominantly distributed in ZI,and were more concentrated in ZIv.The activation of GABAergic neurons in ZIv of DCP group mice provides morphological evidence on the involvement of GABAergic neurons in chronic itch and associated negative emotions.

Objective To establish a method for the simultaneous determination of DOX·HCl and LND. Methods HPLC was performed on Agilent 5 HC-C₁₈（2） （4.6 mm × 250 mm, 5 µm） column. The mobile phase was methanol-0.1% TFA aqueous solution, and the gradient elution procedure were: 0 to 3 min, 65% methanol; 3 to 7 min, 65%→90% methanol; 7 to 13 min, 90% methanol; 13 to 15 min, 90%→65% methanol; 15 to 20 min, 65% methanol. The collection time was 20 min, the balance time was 3 min, the UV detection wavelengths were 205 nm and 253 nm. The flow rate was 1.0 ml/min and the column temperature was 35℃. The amount of inlet was 10 µl. Results The method was highly specific, and both DOX·HCl and LND exhibited good linearity in the concentration range of 1-40 µg/ml and 6-240 µg/ml, respectively. The two compounds’ precision, stability, and recovery satisfied the requirements of the method. Conclusion This study established a HPLC method that was suitable for the simultaneous detection of DOX·HCl and LND. This method’s high level of specificity, accuracy, and reliability .

Objective To observe the value of FOLFOX-hepatic arterial infusion chemotherapy(HAIC)combined with programmed cell death receptor-1(PD-1)inhibitor+targeted drug for treating China liver cancer staging(CNLC)stageⅢa hepatocellular carcinoma(HCC).Methods Sixty-one patients with CNLC stage Ⅲa HCC who underwent PD-1 inhibitor+targeted drug treatment were retrospectively enrolled and divided into observation group(n=30)and control group(n=31)based on whether received FOLFOX-HAIC treatment or not.The general information,treatment strategy,adverse reactions and therapeutic effects were compared between groups,and the value of treatment strategy in observation group was analyzed.Results No significant difference of general information nor PD-1 inhibitor+targeted drug strategy was found between groups(both P＞0.05).Among 1-2 grade adverse reactions,the incidence of nausea,vomiting and abdominal pain in observation group were higher than those in control group(both P＜0.05),while no significant difference of the other 1-2 grade nor 3 grade adverse reactions was observed(all P＞0.05).The objective response rate(ORR),progression free survival(PFS)and overall survival(OS)of observation group were all higher than those of control group(all P＜0.05).Conclusion FOLFOX-HAIC combined with PD-1 inhibitor+targeted drug was more effective for treating CNLC stage Ⅲa HCC with acceptable safety.