ObjectiveTo improve the therapeutic effect of Buyang Huanwutang（BYHW） on diabetic kidney disease （DKD） and explore new methods for developing new Chinese medicine decoctions，we utilized 5-hydroxymethylcytosine （5hmC）-Seal sequencing technology and network pharmacology to modify BYHW. MethodsWe selected 14 diabetes mellitus （DM） patients and 15 DKD patients hospitalized in the Department of Endocrinology of Peking University Third Hospital in 2021. Circulating free DNA （cfDNA） in the patients’ plasma was sequenced. After data processing and screening， we performed temporal clustering analysis to select a DKD 5hmC gene set， which was then cross-validated with a DKD database gene set to obtain the DKD gene set. We retrieved target genes of the seven herbal components of BYHW from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Encyclopedia of Traditional Chinese Medicine （ETCM）， and performed cross-analysis with the DKD gene set to identify common genes shared by the disease and the Chinese medicines. A protein-protein interaction （PPI） network was constructed for the common genes to screen out the key genes. Chinese medicines targeting these key genes were searched against ETCM to identify removable Chinese medicines. Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis was performed on non-common DKD genes， and key genes in DKD-related pathways were selected based on machine learning. The GSE30529 dataset was used to verify the expression trends of 5hmC-modified genes and the feasibility of target genes as drug targets. TCMBank was used to search for target genes and obtain compounds targeting these genes and the corresponding Chinese medicines to construct a "key target-compound-Chinese medicine" network. Molecular docking was employed to verify the binding affinity of compounds with key targets. TCMSP and ETCM were used to search and count the candidate Chinese medicines targeting DKD-related genes， and a new decoction was formed by adding the selected Chinese medicines. A mouse model of DKD was established to examine the efficacy of the new decoction based on the mouse body mass， random blood glucose， urinary microalbumin （mALB）， serum creatinine （Scr）， and blood urea nitrogen （BUN） and by hematoxylin-eosin staining， periodic acid-Schiff staining， Masson staining， immunofluorescence assay， and Real-time PCR. ResultsThe cross-analysis results showed that the DKD gene set included 507 genes， of which 30 were target genes of BYHW. The PPI analysis indicated that the top 15% target genes regarding the degree were interleukin-6 （IL-6）， Toll-like receptor 4 （TLR4）， lactotransferrin （LTF）， lipoprotein lipase （LPL）， and sterol regulatory element-binding transcription factor 1 （SREBF1）. Persicae Semen and Pheretima in BYHW were unrelated to key genes and removed. Machine learning identified 10 potential target genes， among which TBC1 domain family member 5 （TBC1D5）， RAD51 paralog B （RAD51B）， and proteasome 20S subunit alpha 6 （PSMA6） had expression trends consistent with the GSE30529 dataset and could serve as drug targets. The "key target-compound-Chinese medicine" network and molecular docking results indicated that the compounds with good binding affinity to target proteins were arginine， glycine， myristicin， serine， and tyrosine， corresponding to 121 Chinese medicines. The top 10 Chinese medicines targeting DKD-related genes were Lycii Fructus， Ginseng Radix et Rhizoma， Dioscoreae Rhizoma， Rehmanniae Radix Praeparata， Isatidis Radix， Glehniae Radix， Ophiopogonis Radix， Allii Sativi Bulbus， Isatidis Folium， and Bolbostemmatis Rhizoma. Based on traditional Chinese medicine theory， the new decoction was obtained after removal of Persicae Semen and Pheretima and addition of Rehmanniae Radix Praeparata and Dioscoreae Rhizoma. Animal experiment results indicated that the modified BYHW improved the kidney function and inhibited renal fibrosis in DKD mice， with better effects than the original decoction. ConclusionThe BYHW modified based on 5hmC-Seal sequencing demonstrates better performance in inhibiting fibrosis and ameliorating DKD than the original decoction. This elucidates the biomedical theory behind the epigenetic modification of traditional Chinese medicine prescriptions， potentially offering new perspectives for the exploration of these prescriptions

ObjectiveTo improve the therapeutic effect of Buyang Huanwutang（BYHW） on diabetic kidney disease （DKD） and explore new methods for developing new Chinese medicine decoctions，we utilized 5-hydroxymethylcytosine （5hmC）-Seal sequencing technology and network pharmacology to modify BYHW. MethodsWe selected 14 diabetes mellitus （DM） patients and 15 DKD patients hospitalized in the Department of Endocrinology of Peking University Third Hospital in 2021. Circulating free DNA （cfDNA） in the patients’ plasma was sequenced. After data processing and screening， we performed temporal clustering analysis to select a DKD 5hmC gene set， which was then cross-validated with a DKD database gene set to obtain the DKD gene set. We retrieved target genes of the seven herbal components of BYHW from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and the Encyclopedia of Traditional Chinese Medicine （ETCM）， and performed cross-analysis with the DKD gene set to identify common genes shared by the disease and the Chinese medicines. A protein-protein interaction （PPI） network was constructed for the common genes to screen out the key genes. Chinese medicines targeting these key genes were searched against ETCM to identify removable Chinese medicines. Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis was performed on non-common DKD genes， and key genes in DKD-related pathways were selected based on machine learning. The GSE30529 dataset was used to verify the expression trends of 5hmC-modified genes and the feasibility of target genes as drug targets. TCMBank was used to search for target genes and obtain compounds targeting these genes and the corresponding Chinese medicines to construct a "key target-compound-Chinese medicine" network. Molecular docking was employed to verify the binding affinity of compounds with key targets. TCMSP and ETCM were used to search and count the candidate Chinese medicines targeting DKD-related genes， and a new decoction was formed by adding the selected Chinese medicines. A mouse model of DKD was established to examine the efficacy of the new decoction based on the mouse body mass， random blood glucose， urinary microalbumin （mALB）， serum creatinine （Scr）， and blood urea nitrogen （BUN） and by hematoxylin-eosin staining， periodic acid-Schiff staining， Masson staining， immunofluorescence assay， and Real-time PCR. ResultsThe cross-analysis results showed that the DKD gene set included 507 genes， of which 30 were target genes of BYHW. The PPI analysis indicated that the top 15% target genes regarding the degree were interleukin-6 （IL-6）， Toll-like receptor 4 （TLR4）， lactotransferrin （LTF）， lipoprotein lipase （LPL）， and sterol regulatory element-binding transcription factor 1 （SREBF1）. Persicae Semen and Pheretima in BYHW were unrelated to key genes and removed. Machine learning identified 10 potential target genes， among which TBC1 domain family member 5 （TBC1D5）， RAD51 paralog B （RAD51B）， and proteasome 20S subunit alpha 6 （PSMA6） had expression trends consistent with the GSE30529 dataset and could serve as drug targets. The "key target-compound-Chinese medicine" network and molecular docking results indicated that the compounds with good binding affinity to target proteins were arginine， glycine， myristicin， serine， and tyrosine， corresponding to 121 Chinese medicines. The top 10 Chinese medicines targeting DKD-related genes were Lycii Fructus， Ginseng Radix et Rhizoma， Dioscoreae Rhizoma， Rehmanniae Radix Praeparata， Isatidis Radix， Glehniae Radix， Ophiopogonis Radix， Allii Sativi Bulbus， Isatidis Folium， and Bolbostemmatis Rhizoma. Based on traditional Chinese medicine theory， the new decoction was obtained after removal of Persicae Semen and Pheretima and addition of Rehmanniae Radix Praeparata and Dioscoreae Rhizoma. Animal experiment results indicated that the modified BYHW improved the kidney function and inhibited renal fibrosis in DKD mice， with better effects than the original decoction. ConclusionThe BYHW modified based on 5hmC-Seal sequencing demonstrates better performance in inhibiting fibrosis and ameliorating DKD than the original decoction. This elucidates the biomedical theory behind the epigenetic modification of traditional Chinese medicine prescriptions， potentially offering new perspectives for the exploration of these prescriptions

Trials within cohorts （TwiCs） are design methods derived from randomized controlled trials （RCTS）. They have been widely used in chronic disease areas such as tumors and cardiovascular diseases. The basis of the TwiCs design is a prospective cohort of specific diseases. When RCTS need to be implemented， some patients meeting the inclusion and exclusion criteria are randomly sampled from the cohort to receive "trial interventions"， while the remaining patients in the cohort who meet the inclusion and exclusion criteria continue to receive conventional treatment as control groups. By comparing the efficacy differences between the intervention measures of the trial group and the control group， the efficacy of intervention measures was evaluated. Within the cohort， the same process could be repeated to carry out multiple RCTS， so as to evaluate different intervention measures or compare the efficacy of different doses or timing of interventions. Compared with classical RCTS， TwiCs make it easier to recruit patients from the cohort and have higher external validity， providing a new research paradigm for improving the efficiency and applicability of RCTS in clinical practice. However， TwiCs may also face the challenge of poor compliance of patients in the cohort. Researchers need to take effective measures to control these patients in the design and operation of TwiCs. This article focused on the methodological key points during the implementation of TwiCs， including multi-stage informed consent （patients are informed of consent at three stages： entering the cohort， entering the trial group， and after the trial）， randomization procedures （only random sampling of patients from the cohort to receive "trial interventions"）， sample size calculation， and statistical analysis methods. The article also compared the differences between TwiCs and traditional RCTS and illustrated TwiCs research design and analysis with examples， so as to provide new research ideas and methods for clinical researchers.

BACKGROUND:Diabetic osteoporosis is gaining public attention.However,few studies have reported the effect of a high-glucose environment on the osteogenic differentiation of human umbilical cord mesenchymal stem cells and the corresponding therapeutic strategies. OBJECTIVE:To investigate whether vitamin D3 can restore the osteogenic differentiation potential of human umbilical cord mesenchymal stem cells in a high-glucose environment. METHODS:The viability of human umbilical cord mesenchymal stem cells was detected by CCK-8 assay to screen the appropriate vitamin D3 intervention concentration.Under the high-glucose environment,RT-qPCR,western blot assay,immunofluorescence,JC-1 mitochondrial membrane potential,alizarin red staining,and β-galactosidase staining were used to evaluate the osteogenic differentiation potential,intracellular reactive oxygen species accumulation,mitochondrial membrane potential alteration,and cell senescence of human umbilical cord mesenchymal stem cells after vitamin D3 intervention.The underlying mechanism was also discussed. RESULTS AND CONCLUSION:(1)Vitamin D3 significantly promoted the proliferation of human umbilical cord mesenchymal stem cells in the range of 0.1 μmol/L to 1 mmol/L.(2)High-glucose environment down-regulated the mRNA and protein level expressions of osteogenic-related genes α1-I collagen,alkaline phosphatase,Runt-associated transcription factor 2,and osteocalcin in human umbilical cord mesenchymal stem cells,which induced oxidative stress and cellular senescence.(3)Vitamin D3 at an intervention concentration of 10 μmol/L significantly restored the osteogenic phenotype of human umbilical cord mesenchymal stem cells under high-glucose conditions and attenuated intracellular oxidative stress and cellular senescence by activating the Nrf2/HO-1 signaling pathway.(4)These findings suggested that the osteogenic differentiation ability of human umbilical cord mesenchymal stem cells was reduced in the high-glucose environment,and vitamin D3 could partially improve their osteogenic differentiation ability and reduce cell damage.

Objective:To investigate the relationship between serum levels of interleukin-17A (IL-17A) and chemokine ligand 19 (CCL19) and disease activity in patients with lupus nephritis.Methods:A total of 100 patients with lupus nephritis admitted to Affiliated Hospital of Jining Medical College from June 2020 to February 2023 were collected as the disease group, according to the disease activity index, patients were grouped into inactive group (32 cases), mild active group (21 cases), moderate active group (29 cases), and severe active group (18 cases); another 100 healthy individuals who underwent physical examinations in our hospital during the same period were collected as the control group. Enzyme linked immunosorbent assay (ELISA) was applied to detect the expression levels of IL-17A and CCL19 in serum; Pearson method was applied to analyze the correlation between serum IL-17A, CCL19 and routine indicators in patients with lupus nephritis; receiver operating characteristic curve was applied to analyze the diagnostic value of serum IL-17A and CCL19 for moderate/severe lupus nephritis disease activity.Results:The expression levels of IL-17A and CCL19 in the serum of the disease group were obviously higher than those of the control group: (252.63 ± 64.47) ng/L vs. (123.27 ± 25.12) ng/L and (566.98 ± 73.36) ng/L vs. (275.63 ± 50.48) ng/L ( t = 18.70 and 32.72, P<0.05); the serum levels of IL-17A and CCL19 in the severe active, moderate active, and mild active groups were higher than those in the inactive group: (331.42 ± 87.46), (278.50 ± 74.19) and (232.34 ± 59.16) ng/L vs. (198.18 ± 46.22) ng/L; (662.33 ± 89.57), (606.14 ± 79.25) and (552.84 ± 68.36) ng/L vs. (487.13 ± 62.19) ng/L, and with the increase of disease activity, the levels of serum IL-17A and CCL19 gradually increased ( F = 17.86 and 25.35, P<0.05); the glomerular filtration rate, albumin, complement C 3 and complement C 4 in the active group were obviously lower than those in the inactive group: (69.17 ± 13.25) ml/(min·1.73 m 2) vs. (86.18 ± 14.16) ml/(min·1.73 m 2), (24.18 ± 5.11) g/L vs. (31.25 ± 6.35) g/L, (432.35 ± 95.22) mg/L vs. (675.42 ± 125.16) mg/L, (76.58 ± 17.51) mg/L vs. (121.42 ± 27.18) mg/L, while blood creatinine, urine protein and erythrocyte sedimentation rate were obviously higher than those in the inactive group: (92.34 ± 16.24) μmoI/L vs. (53.21 ± 9.17) μmoI/L, (3.43 ± 0.82) g/24 h vs. (1.26 ± 0.23) g/24 h, (66.37 ± 12.28) mm/1 h vs. (35.62 ± 8.67) mm/1 h ( t = 5.86, 5.97, 10.74, 9.93, 12.70, 14.67 and 12.74; P<0.05); serum IL-17A and CCL19 in patients with lupus nephritis were negatively correlated with glomerular filtration rate, albumin, complement C 3, and complement C 4, while positively correlated with blood creatinine, urine protein, and ESR ( P<0.05); the area under the curve (AUC) of the combined diagnosis of serum IL-17A and CCL19 for lupus nephritis disease activity was 0.961, which was superior to their respective individual diagnoses ( Z = 2.24 and 3.16, P = 0.025 and 0.002). Conclusions:The expression levels of IL-17A and CCL19 in serum gradually increase with the increase of disease activity in patients with lupus nephritis. The combined detection of the two has good diagnostic value for disease activity in lupus nephritis.

Objective To explore the efficacy and safety of enhanced external counterpulsation(EECP)in elderly patients with acute ischemic stroke(AIS)complicated by coronary heart dis-ease(CHD).Methods A total of 65 AIS patients with CHD admitted in our hospital from Janu-ary to June 2023 were recruited and randomly divided into a control group(drug secondary pre-vention,n=32)and a treatment group(drug combined with EECP therapy,n=33).Their NIHSS score,mRS score and Canadian Cardiovascular Society(CCS)angina grade were evaluated before and after treatment and compared between the two groups.The incidences of recurrent ischemic stroke,new hemorrhagic stroke and major adverse cardiovascular events(MACE)were also recor-ded during treatment.Results The NIHSS score and mRS score were significantly decreased in both groups after treatment(P＜0.01).After treatment,the NIHSS score(2.67±1.63 vs 3.56± 1.83),mRS score[1.0(0.0,1.0)vs 2.0(1.0,2.0)]and CCS grade[1.0(1.0,2.0)vs 2.0(1.0,2.0)]were obviously lower in the treatment group than the control group(P＜0.05,P＜0.01).There were no statistical differences in the incidence rates of recurrent ischemic stroke,new-onset hem-orrhagic stroke,and MACE between the control group and the treatment group(9.4％vs 6.1％,6.3％vs 3.0％,12.5％vs 6.1％,P＞0.0 5).Conclusion EECP is a safe and effective treatment option for elderly AIS patients with CHD.

Objective:To analyze the short-term clinical outcomes of total laparoscopic distal gastrectomy (TLDG) and laparoscopic-assisted distal gastrectomy (LADG) combined with Billroth-Ⅱ+Braun anastomosis.Methods:Clinical characteristics of patients undergoing laparoscopic distal gastrectomy combined with Billroth-Ⅱ+Braun anastomosis at Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University from Jan 2020 to Oct 2022 were analyzed. Patients were divided into TLDG group ( n=62) and LADG group ( n=62) according to the surgical approach. Results:There were significant differences in the preoperative clinical data section between the two groups, and 124 patients (62 in each group) were enrolled after using propensity score matching to balance significant variables. Compared with the LADG group, the TLDG group showed statistically differences in time to first venting [(2.9±1.3) vs. (2.3±0.8) d, Z=-3.072, P=0.002], time to first fluid diet [(5.9±1.3) vs. (5.4±1.4) d, Z=-2.031, P=0.042] and incision length [(7.1±1.4) vs. (4.8±0.8) cm, Z=-6.331, P=0.000]. Total postoperative complication rate in the TLDG group and the LADG group (29% vs. 37%, χ2=0.911, P=0.340) was not statistically significant. Incidence of postoperative pneumonia was lower in the TLDG group than in the LADG group (3% vs. 13%, χ2=3.916, P=0.048), and incidence of all remaining postoperative complications were not statistically significant. There was no statistically significant difference in the incidence of serious postoperative complications between the TLDG and LADG groups ( P=1.000). Multifactorial analysis revealed that male ( P=0.023) and age ≥65 years ( P=0.001) were independent risk factors for postoperative complications. Conclusion:TLDG is safe and feasible and has better short-term clinical efficacy than LADG.

Oral diseases concern almost every individual and are a serious health risk to the popula-tion.The restorative treatment of tooth and jaw defects is an important means to achieve oral function and support the appearance of the contour.Based on the principle of"learning from the nature",Deng Xu-liang's group of Peking University School and Hospital of Stomatology has proposed a new concept of"microstructural biomimetic design and tissue adaptation of tooth/jaw materials"to address the worldwide problems of difficulty in treating dentine hypersensitivity,poor prognosis of restoration of tooth defects,and vertical bone augmentation of alveolar bone after tooth loss.The group has broken through the bottle-neck of multi-stage biomimetic technology from the design of microscopic features to the enhancement of macroscopic effects,and invented key technologies such as crystalline/amorphous multi-level assembly,ion-transportation blocking,and multi-physical properties of the micro-environment reconstruction,etc.The group also pioneered the cationic-hydrogel desensitizer,digital stump and core integrated restora-tions,and developed new crown and bridge restorative materials,gradient functionalisation guided tissue regeneration membrane,and electrically responsive alveolar bone augmentation restorative membranes,etc.These products have established new clinical strategies for tooth/jaw defect repair and achieved inno-vative results.In conclusion,the research results of our group have strongly supported the theoretical im-provement of stomatology,developed the technical system of oral hard tissue restoration,innovated the clinical treatment strategy,and led the progress of the stomatology industry.

BACKGROUND:Osteoporosis has a high incidence,leading to fracture and other complications.However,existing drugs have great side effects and are difficult to meet the clinical application. OBJECTIVE:To explore the effect and potential mechanism of fucoxanthin on osteoporosis induced by glucocorticoid. METHODS:Primary rat osteoblasts were inoculated in 6-well plates.When the cell fusion reached 80%,the cells were divided into four groups:the control group was cultured alone for 24 hours,the glucocorticoid group was intervened with dexamethasone for 24 hours,the fucoxanthin group was intervened with fucoxanthin for 24 hours,and the glucocorticoid + fucoxanthin group was intervened with dexamethasone and fucoxanthin at the same time for 24 hours.After intervention,cell proliferation,apoptosis,intracellular reactive oxygen species level,and protein expression of apoptosis-related proteins,bone formation-related proteins,and nuclear factor erythroid-2-related factor 2 were detected. RESULTS AND CONCLUSION:Cell counting kit-8 results showed that the cell viability was decreased in the glucocorticoid group compared with the control group(P＜0.05)but increased in the glucocorticoid+fucoxanthin group compared with the glucocorticoid group(P＜0.05).JC-1 mitochondrial membrane potential staining and flow cytometry assay showed that the percentage of apoptosis increased in the glucocorticoid group compared with the control group(P＜0.05)but decreased in the glucocorticoid+fucoxanthin group compared with the glucocorticoid group(P＜0.05).Western blot assay showed that compared with the control group,the protein expression of BAX and cleaved poly(ADP-ribose)polymerase was elevated in the glucocorticoid group(P＜0.05),and the protein expression of BCL2,type Ⅰ collagen α1 peptide chain,alkaline phosphatase,osteocalcin,and RUNX2 was decreased in the glucocorticoid group(P＜0.05).Compared with the glucocorticoid group,the protein expression of BAX and cleaved poly(ADP-ribose)polymerase was decreased(P＜0.05),and the protein expression of BCL2,type Ⅰ collagen α1 peptide chain,alkaline phosphatase,osteocalcin,and RUNX2 was elevated(P＜0.05)in the glucocorticoid+fucoxanthin group.Fluorescent probe assay showed an increase in reactive oxygen species level in the glucocorticoid group compared with the control group(P＜0.05)and a decrease in reactive oxygen species level in the glucocorticoid+fucoxanthin group compared with the glucocorticoid group(P＜0.05).Immunofluorescence staining and western blot assay showed that the protein expression of nuclear factor erythroid-2-related factor 2 in the glucocorticoid group was decreased compared with that in the control group(P＜0.05);and the protein expression of nuclear factor erythroid-2-related factor 2 in the glucocorticoid+fucoxanthin group was elevated compared with that in the glucocorticoid group(P＜0.05).To conclude,fucoxanthin can improve glucocorticoid-induced osteoblast apoptosis and the expression of bone formation-related molecules by activating nuclear factor erythroid-2-related factor 2.

A model eye is used to study the anatomical structure and optical properties of the eye, and to analyze visual quality under different conditions using optical analysis software. By adjusting biological parameters such as axial length and corneal curvature radius and modifying its refractive state, the model eye can be used for ophthalmic surgical planning and treatment program development, leading to more accurate and objective assessments of visual quality. While previously used mainly for theoretical studies in optics and ophthalmology, model eyes and optical analysis software are now being applied to various ocular diseases, with their accuracy confirmed through comparison with clinical trials. This article aims to summarize the widely used model eyes and optical analysis software, as well as their applications, to offer new perspectives for diagnosing and treating ocular diseases and evaluating visual quality.