Objective To explore the mechanism of Danggui Sini Decoction in treating knee osteoarthritis(KOA)by using network pharmacology,combined with in vitro experimental verification.Methods The active components of Danggui Sini Decoction were retrieved and screened through TCMSP,CNKI and PubMed databases,and their corresponding targets were predicted using the SwissTarget Prediction platform.KOA related targets were retrieved from GeneCards and OMIM databases.Intersection of drug targets and KOA targets was obtained,and key components,core targets,and their related biological mechanisms were identified through network topology analysis,GO and KEGG pathway enrichment analysis.Molecular docking was used to verify the degree of binding between key components and core targets.Danggui Sini Decoction containing serum was prepared,and an in vitro KOA model was constructed by inducing rat articular chondrocytes with lipopolysaccharide.The CCK-8 method was used to screen for the optimal concentration of drug containing serum intervention,fluorescent probes were used to detect intracellular ROS content,immunofluorescence was used to detect NF-κB p65 nuclear translocation,RT-qPCR was used to detect the expressions of IL-6,IL-1β and TNF-α mRNA,Western blot was used to detect the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),NF-κB p65 and p-NF-κB p65 proteins.Results The results of network pharmacology analysis indicated that the treatment of KOA with Danggui Sini Decoction may involve regulating Toll like receptors,MAPK,TNF,apoptosis and other inflammatory and aging signaling pathways,compounds such as quercetin,kaempferol and glycyrrhizin may play important roles.Core targets included IL6,IL1B,TNF,TLR4,NFKB1,JUN,MAPK1,RELA.In vitro experiments showed that compared with the blank group,the ROS content in chondrocytes of the model group significantly increased(P<0.01),NF-κB p65 protein was significantly nuclear translocation(P<0.01),and mRNA expressions of IL-6,IL-1β,TNF-α mRNA and protein expressions of TLR4,MyD88 and p-NF-κB p65 significantly increased(P<0.01);compared with the model group,the intervention of Danggui Sini Decoction containing serum could significantly reduce intracellular ROS content(P<0.05),inhibit NF-κB p65 nuclear translocation(P<0.01),and reduce the expression of inflammatory factor mRNA and related proteins(P<0.05,P<0.01).Conclusion Danggui Sini Decoction can significantly reduce the inflammatory response of rat articular chondrocytes induced by lipopolysaccharide and exert therapeutic effects on KOA.Its mechanism may be related to the inhibition of TLR4/MyD88/NF-κB pathway.

Objective To explore the characteristics of incorrect opinions in re-examination cases,analyze the reasons,characteristics,and commonalities of different forensic opinions in the same injury case during re-examination,and provide references for similar case acceptance,forensic procedures,analysis and argumentation,and the application of relevant provisions.Methods A retrospective analysis was conducted on the re-examination cases of human injury degree accepted by the Forensic Judicial Appraisal Center of Guizhou Medical University and West China Forensic Medical Appraisal Center of Sichuan University from January 2020 to December 2024.Results Among the 81 collected re-examination cases of human injury degree,the injury types mainly included fractures,soft tissue injuries,tendon injuries,craniocerebral injuries,nerve injuries,joint injuries,and organ injuries.Of these,74 cases were entrusted by the public security system,and 72 cases used primary injuries as the basis for assessment.Seven cases had more than 3 appraisals and 74 cases had two appraisals,of which 53 cases had inconsistent opinions between the previous and current appraisals,including 29 cases with an upgraded injury grade and 24 cases with a downgraded injury grade.Conclusion When accepting re-examination cases,forensic experts need to choose relevant objective examination techniques in forensic clinical medicine,such as clinical imaging techniques,to determine whether imaging data serves as key evidence in the case,and seek assistance from experts in related fields when necessary.In the case acceptance stage,it is necessary to collect and review key assessment materials such as medical records,the first assessment opinion,and interrogation records.Forensic experts should also enhance their understanding of legal provisions,timing of assessment,their image-reading ability,and work to standardize the forensic assessment process.

Objective To explore the mechanism of Danggui Sini Decoction in treating knee osteoarthritis(KOA)by using network pharmacology,combined with in vitro experimental verification.Methods The active components of Danggui Sini Decoction were retrieved and screened through TCMSP,CNKI and PubMed databases,and their corresponding targets were predicted using the SwissTarget Prediction platform.KOA related targets were retrieved from GeneCards and OMIM databases.Intersection of drug targets and KOA targets was obtained,and key components,core targets,and their related biological mechanisms were identified through network topology analysis,GO and KEGG pathway enrichment analysis.Molecular docking was used to verify the degree of binding between key components and core targets.Danggui Sini Decoction containing serum was prepared,and an in vitro KOA model was constructed by inducing rat articular chondrocytes with lipopolysaccharide.The CCK-8 method was used to screen for the optimal concentration of drug containing serum intervention,fluorescent probes were used to detect intracellular ROS content,immunofluorescence was used to detect NF-κB p65 nuclear translocation,RT-qPCR was used to detect the expressions of IL-6,IL-1β and TNF-α mRNA,Western blot was used to detect the expression of Toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88),NF-κB p65 and p-NF-κB p65 proteins.Results The results of network pharmacology analysis indicated that the treatment of KOA with Danggui Sini Decoction may involve regulating Toll like receptors,MAPK,TNF,apoptosis and other inflammatory and aging signaling pathways,compounds such as quercetin,kaempferol and glycyrrhizin may play important roles.Core targets included IL6,IL1B,TNF,TLR4,NFKB1,JUN,MAPK1,RELA.In vitro experiments showed that compared with the blank group,the ROS content in chondrocytes of the model group significantly increased(P<0.01),NF-κB p65 protein was significantly nuclear translocation(P<0.01),and mRNA expressions of IL-6,IL-1β,TNF-α mRNA and protein expressions of TLR4,MyD88 and p-NF-κB p65 significantly increased(P<0.01);compared with the model group,the intervention of Danggui Sini Decoction containing serum could significantly reduce intracellular ROS content(P<0.05),inhibit NF-κB p65 nuclear translocation(P<0.01),and reduce the expression of inflammatory factor mRNA and related proteins(P<0.05,P<0.01).Conclusion Danggui Sini Decoction can significantly reduce the inflammatory response of rat articular chondrocytes induced by lipopolysaccharide and exert therapeutic effects on KOA.Its mechanism may be related to the inhibition of TLR4/MyD88/NF-κB pathway.

Objective To explore the characteristics of incorrect opinions in re-examination cases,analyze the reasons,characteristics,and commonalities of different forensic opinions in the same injury case during re-examination,and provide references for similar case acceptance,forensic procedures,analysis and argumentation,and the application of relevant provisions.Methods A retrospective analysis was conducted on the re-examination cases of human injury degree accepted by the Forensic Judicial Appraisal Center of Guizhou Medical University and West China Forensic Medical Appraisal Center of Sichuan University from January 2020 to December 2024.Results Among the 81 collected re-examination cases of human injury degree,the injury types mainly included fractures,soft tissue injuries,tendon injuries,craniocerebral injuries,nerve injuries,joint injuries,and organ injuries.Of these,74 cases were entrusted by the public security system,and 72 cases used primary injuries as the basis for assessment.Seven cases had more than 3 appraisals and 74 cases had two appraisals,of which 53 cases had inconsistent opinions between the previous and current appraisals,including 29 cases with an upgraded injury grade and 24 cases with a downgraded injury grade.Conclusion When accepting re-examination cases,forensic experts need to choose relevant objective examination techniques in forensic clinical medicine,such as clinical imaging techniques,to determine whether imaging data serves as key evidence in the case,and seek assistance from experts in related fields when necessary.In the case acceptance stage,it is necessary to collect and review key assessment materials such as medical records,the first assessment opinion,and interrogation records.Forensic experts should also enhance their understanding of legal provisions,timing of assessment,their image-reading ability,and work to standardize the forensic assessment process.

Objective:To investigate the long-term outcomes of minimally invasive Oxford phase Ⅲ unicompartmental knee arthroplasty (UKA) for patients with medial compartment osteoarthropathy.Methods:The clinical data of 594 patients (701 knees) who underwent minimally invasive UKA with Oxford phase Ⅲ unicompartmental prosthesis at Department of Orthopedics,the Affiliated Hospital of Qingdao University from January 2007 to January 2016 were retrospectively analyzed.There were 155 males and 439 females,aged (62.6±10.9) years (range: 44 to 81 years),with a body mass index of (26.9±3.8) kg/m 2 (range: 21.1 to 36.2 kg/m 2).There were 359 left knees and 342 right knees,676 knees with osteoarthritis and 25 knees with idiopathic osteonecrosis of the medial femoral condyle.There were 487 cases underwent UKA (66 cases underwent UKA on one side and total knee arthroplasty on the other) and 107 cases underwent bilateral UKA.Patients′ prosthetic survival,complications,range of motion(ROM) of the knee,visual analogue score (VAS),Western Ontario and McMaster University (WOMAC) osteoarthritis index,and American knee society score (KSS) were collected to assess clinical outcomes.Paired sample t test was used to compare the data before and after operation. Results:All patients completed the surgery successfully.There was no intraoperative fractures,postoperative infections or symptomatic vascular embolic disease occurred.The postoperative complications,including mobile bearing dislocation,prosthesis loosening,tibial plateau collapse,the lateral compartment degeneration and postoperative pain were occurred in 18 cases (3.0%,18/594).Thirteen patients suffered complications were transferred to total knee arthroplasty,4 underwent partial revision,if this was used as the endpoint of the study,the surgical success rate was 97.1% (577/594) and the prosthetic revision rate was 2.9%.The ROM was improved from(105.9±11.8)°preoperatively to (114.0±13.3)° at the last follow-up ( t=10.796, P<0.01);the KSS clinical score was increased from 54.3±3.6 to 90.1±6.0 ( P<0.01) and the functional score was increased from 55.9±3.9 to 87.5±5.7( t=124.325, P<0.01; t=110.985, P<0.01).The WOMAC osteoarthritis index was decreased from 54.8±6.7 to 9.2±3.1 at the last follow-up( t=150.860, P<0.01) and the VAS was decreased from 6.1±1.1 to 1.5±1.0 at the last follow-up( t=74.941, P<0.01). Conclusions:Minimally invasive Oxford phase Ⅲ UKA for medial compartment knee osteoarthritis has a favorable prosthesis survival rate,low revision rate,and few complications at long-term follow-up.Patients have significant improvement in knee function with satisfactory clinical outcomes.

Objective:To investigate the long-term outcomes of minimally invasive Oxford phase Ⅲ unicompartmental knee arthroplasty (UKA) for patients with medial compartment osteoarthropathy.Methods:The clinical data of 594 patients (701 knees) who underwent minimally invasive UKA with Oxford phase Ⅲ unicompartmental prosthesis at Department of Orthopedics,the Affiliated Hospital of Qingdao University from January 2007 to January 2016 were retrospectively analyzed.There were 155 males and 439 females,aged (62.6±10.9) years (range: 44 to 81 years),with a body mass index of (26.9±3.8) kg/m 2 (range: 21.1 to 36.2 kg/m 2).There were 359 left knees and 342 right knees,676 knees with osteoarthritis and 25 knees with idiopathic osteonecrosis of the medial femoral condyle.There were 487 cases underwent UKA (66 cases underwent UKA on one side and total knee arthroplasty on the other) and 107 cases underwent bilateral UKA.Patients′ prosthetic survival,complications,range of motion(ROM) of the knee,visual analogue score (VAS),Western Ontario and McMaster University (WOMAC) osteoarthritis index,and American knee society score (KSS) were collected to assess clinical outcomes.Paired sample t test was used to compare the data before and after operation. Results:All patients completed the surgery successfully.There was no intraoperative fractures,postoperative infections or symptomatic vascular embolic disease occurred.The postoperative complications,including mobile bearing dislocation,prosthesis loosening,tibial plateau collapse,the lateral compartment degeneration and postoperative pain were occurred in 18 cases (3.0%,18/594).Thirteen patients suffered complications were transferred to total knee arthroplasty,4 underwent partial revision,if this was used as the endpoint of the study,the surgical success rate was 97.1% (577/594) and the prosthetic revision rate was 2.9%.The ROM was improved from(105.9±11.8)°preoperatively to (114.0±13.3)° at the last follow-up ( t=10.796, P<0.01);the KSS clinical score was increased from 54.3±3.6 to 90.1±6.0 ( P<0.01) and the functional score was increased from 55.9±3.9 to 87.5±5.7( t=124.325, P<0.01; t=110.985, P<0.01).The WOMAC osteoarthritis index was decreased from 54.8±6.7 to 9.2±3.1 at the last follow-up( t=150.860, P<0.01) and the VAS was decreased from 6.1±1.1 to 1.5±1.0 at the last follow-up( t=74.941, P<0.01). Conclusions:Minimally invasive Oxford phase Ⅲ UKA for medial compartment knee osteoarthritis has a favorable prosthesis survival rate,low revision rate,and few complications at long-term follow-up.Patients have significant improvement in knee function with satisfactory clinical outcomes.

BACKGROUND: How to effectively and rapidly induce the osteogenic differentiation of human umbilical cord mesenchymal stem cells is the focus of the current stem cell research. Increasing evidence has demonstrated some growth factors, such as bone morphogenetic protein-2, have important effects on the transdifferentiation of umbilical cord mesenchymal stem cels into osteoblasts in vitro. However, widespread use of growth factors is limited because of high cost. Insulin is widely used in the cell culture and induction, but there is no report about the effect of insulin on the osteogenic differentiation of human umbilical cord mesenchymal stem cells. OBJECTIVE:To observe the effect of insulin on osteogenic differentiation of human umbilical cord mesenchymal stem cels and to explore the feasibility of human umbilical cord mesenchymal stem cell transplantation in the treatment of diabetic delayed fracture healing. METHODS:The passage 3 human umbilical cord mesenchymal stem cells were inoculated in two flasks, denoted as experimental group and control group. The insulin (10-7mmol/L) was added to the experimental group but not to the control group. The proliferative capacity of human umbilical cord mesenchymal stem cels was evaluated by cell count kit-8 and alkaline phosphatase activity. The osteogenic differentiation capacity of human umbilical cord mesenchymal stem cells was evaluated by measuring the protein and mRNA expressions of type I colagen as well as osteocalcin mRNA level. RESULTS AND CONCLUSION: After 1-2 weeks of induction, compared with the control group, insulin could significantly increase the number of human umbilical cord mesenchymal stem cells in the experimental group, the activity of alkaline phosphatase and expressions of type I collagen osteocalcin mRNA (P< 0.05). These data indicate that insulin can promote the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells.

Objective　To investigate the effects of isocarbophos, a new and highly effective organophosphorus pesticide on the membranes of keratinocytes. Methods　The epidermic cells of human skin cultured in the medium were adopted as a biological material.The activity of lactate dehydrogenase (LDH) was used as an indicator. We tested the activities of LDH and alkaline phosphatase (ALP) in the medium in different isocarbophos dosage conditions. Results　It was found that the activity of LDH in the medium decreased with the increase of isocarbophos dosage. The LDH activities of the dosage groups were statistically different from those of the positive and negative groups. The ALP activities were of no significant difference between the groups. Conclusion　The above findings reveal that isocarbophos does not damage the membranes of keratinocytes directly, howerer it inhibits the LDH of human skin keratinocytes; thus it is suggested that isocarbophos like most organophosphorus pesticides used widely can pass through the intact human skin and poison the body before the skin is damaged. Therefore, protection during work hours is of paramount importance.