Radiation-induced thrombocytopenia (RIT) faces a perplexing challenge in the clinical treatment of cancer patients, and current therapeutic approaches are inadequate in the clinical settings. In this research, oxymatrine, a new molecule capable of healing RIT was screened out, and the underlying regulatory mechanism associated with magakaryocyte (MK) differentiation and thrombopoiesis was demonstrated. The capacity of oxymatrine to induce MK differentiation was verified in K-562 and Meg-01 cells in vitro. The ability to induce thrombopoiesis was subsequently demonstrated in Tg (cd41:enhanced green fluorescent protein (eGFP)) zebrafish and RIT model mice. In addition, we carried out network pharmacological prediction, drug affinity responsive target stability assay (DARTS) and cellular thermal shift assay (CETSA) analyses to explore the potential targets of oxymatrine. Moreover, the pathway underlying the effects of oxymatrine was determined by Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, Western blot (WB), and immunofluorescence. Oxymatrine markedly promoted MK differentiation and maturation in vitro. Moreover, oxymatrine induced thrombopoiesis in Tg (cd41:eGFP) zebrafish and accelerated thrombopoiesis and platelet function recovery in RIT model mice. Mechanistically, oxymatrine directly binds to toll-like receptor 2 (TLR2) and further regulates the downstream pathway stimulator of interferon genes (STING)/nuclear factor-kappaB (NF-κB), which can be blocked by C29 and C-176, which are specific inhibitors of TLR2 and STING, respectively. Taken together, we demonstrated that oxymatrine, a novel TLR2 agonist, plays a critical role in accelerating MK differentiation and thrombopoiesis via the STING/NF-κB axis, suggesting that oxymatrine is a promising candidate for RIT therapy.

Objective To develop a reagent for detecting of 17α-hydroxyprogesterone(17α-OHP)in dried blood spots on filter paper fixed on vascular stent by Auto TRFIA-4 automatic fluorescence immunoanalyzer and evaluate its performance.Methods The microwell plate was coated with the sheep anti-rabbit IgG antibody as microwell reaction plate,the rabbit anti-human 17α-OHP antibody was diluted as intermediate antibody,and the 17α-OHP-bovine serum albumin conjugate was labeled by europium as europium marker.The concentration of 17α-OHP in dried blood spots on filter paper fixed on blood spots stent was quantitatively detected by Auto TRFIA-4 automatic fluorescence immunoanalyzer.The analysis sensitivity,accuracy,linearity,precision,specificity and stability were evaluated,and whether they met the requirements of the formulated industry standards were evaluated.A total of 227 neonatal heel blood filter paper samples from newborns who were born 72 hours after birth and within 7 days and fully breastfeeding were selected for reagent comparison test.The consistency was analyzed by χ2 test,Kappa test,t test,linear correlation analysis,regression analysis,Bland-Altman method analysis and predictive bias analysis of medical decision level,P<0.05 indicated statistically significant difference.Results The optimal coating concentration of sheep anti-rabbit IgG antibody was 3 μg/ml.The optimal dilution ratio of rabbit anti-human 17α-OHP antibody was 1∶1 500.The optimal dilution ratio of 17α-OHP europium marker mother liquor was 1∶2 500.The limit of blank,limit of detection,limit of quantification was 0.75,1.08 and 1.99 nmol/L,respectively.The relative deviations of the standard check test were within±15.00%,and the average recovery rate was 92.36%.The linear correlation coefficient was 0.997 1 in the range of 2.00 to 300.00nmo/L.The intra-assay and inter-assay coefficients of variation were all within 10.00%.The cross-reactivity rates of 100.00 ng/ml progesterone,17α-hydroxypreg nenolone and 11-deoxycortisol were within 0.089%to 0.64%.The performance of stability test met the requirements.The total coincidence rate was 100%compared with the results of contrast reagent.The quantitative results were highly correlated with the contrast reagent(r=0.999 4,tr=452.02,P<0.05).Conclusion The self-developed reagent has the advantages of high sensitivity,good accuracy,wide linear range,good precision,high specificity and good stability,which meets the requirements of the formulated industry standards,and has high correlation and consistency with the result of contrast reagent,which meets the needs of clinical detection.

Radiation-induced thrombocytopenia(RIT)faces a perplexing challenge in the clinical treatment of cancer patients,and current therapeutic approaches are inadequate in the clinical settings.In this research,oxy-matrine,a new molecule capable of healing RIT was screened out,and the underlying regulatory mecha-nism associated with magakaryocyte(MK)differentiation and thrombopoiesis was demonstrated.The capacity of oxymatrine to induce MK differentiation was verified in K-562 and Meg-01 cells in vitro.The ability to induce thrombopoiesis was subsequently demonstrated in Tg(cd41:enhanced green fluorescent protein(eGFP))zebrafish and RIT model mice.In addition,we carried out network pharmacological pre-diction,drug affinity responsive target stability assay(DARTS)and cellular thermal shift assay(CETSA)analyses to explore the potential targets of oxymatrine.Moreover,the pathway underlying the effects of oxymatrine was determined by Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses,Western blot(WB),and immunofluorescence.Oxymatrine markedly promoted MK differentiation and maturation in vitro.Moreover,oxymatrine induced thrombopoiesis in Tg(cd41:eGFP)zebrafish and accelerated thrombopoiesis and platelet function recovery in RIT model mice.Mechanistically,oxymatrine directly binds to toll-like receptor 2(TLR2)and further regulates the downstream pathway stimulator of interferon genes(STING)/nuclear factor-kappaB(NF-κB),which can be blocked by C29 and C-176,which are specific inhibitors of TLR2 and STING,respectively.Taken together,we demonstrated that oxymatrine,a novel TLR2 agonist,plays a critical role in accelerating MK differentiation and thrombopoiesis via the STING/NF-κB axis,suggesting that oxymatrine is a promising candidate for RIT therapy.

Objective To investigate the diagnostic value of serum bromodomain-containing pro-tein 4(BRD4)combined with gastrin-17(G-17)in Helicobacter pylori(Hp)-positive early gastric cancer.Methods A total of 88 patients with Hp-positive early gastric cancer admitted to our hospital from September 2021 to September 2023 were selected as early gastric cancer group.Meanwhile,92 patients with Hp-positive precancerous lesions and 80 patients with Hp-positive gastritis admitted dur-ing the same period were selected as precancerous lesion group and gastritis group,respectively.En-zyme-linked immunosorbent assay(ELISA)was used to detect the serum levels of BRD4 and G-17.Multivariate logistic regression analysis was performed to screen the influencing factors of Hp-positive early gastric cancer.Receiver operating characteristic(ROC)curve analysis was used to evaluate the diagnostic value of serum BRD4 and G-17 in Hp-positive early gastric cancer.Results Compared with the gastritis group,the levels of BRD4 and G-17 in the precancerous lesion group and early gastric cancer group were significantly increased(P＜0.05).Furthermore,compared with the precancerous le-sion group,the levels of BRD4 and G-17 in the early gastric cancer group were also significantly elevated(P＜0.05).The proportion of patients with a family history of gastric cancer,those who preferred hot food,cold food,or high-salt food,as well as the levels of PG Ⅱ,BRD4,and G-17 were signifi-cantly higher in the early gastric cancer group than in the non-gastric cancer group,while the level of pepsinogen(PG)Ⅰ was significantly lower(P＜0.05).Logistic regression analysis revealed that preference for hot food,high-salt food,PG Ⅱ,BRD4,G-17,and PG Ⅰ were all influencing factors for Hp-positive early gastric cancer(P＜0.05).The area under the curve(AUC)values for serum BRD4,G-17,and their combination in diagnosing Hp-positive early gastric cancer were 0.793,0.830,and 0.912,respectively.The diagnostic efficacy of the combined detection was superior to that of single detection(P＜0.05).Conclusion The serum levels of BRD4 and G-17 are elevated in patients with Hp-positive early gastric cancer,and both exhibit certain diagnostic value for Hp-positive early gastric cancer,suggesting their potential as serum biomarkers for the diagnosis of Hp-positive early gastric cancer.

Objective:To explore the safety and effectiveness of ileal bypass with stent to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.Methods:From Dec 2021 to Sep 2024, the clinical data of 55 patients undergoing robot anus-preserving surgery and stent ileal bypass for low rectal cancer in the First Affiliated Hospital of Ningbo University was analyzed retrospectively.Results:All the 55 patients successfully completed the operation. Among them, 47 patients underwent low anterior resection and 8 patients did sphincter resection. The total operation time was (207±37)min, the operation time of ileal bypass was (24±3)min, and the intraoperative blood loss was (32±18)ml. After operation, the time of stent disintegration was (23.93± 2.87) days, and the time of catheter removal was (29.55 ± 3.21) days. By postoperative TNM staging, there were 12 cases in T1 stage, 26 cases in T2 and 17 cases in T3. The number of lymph node dissection was (16.84±4.50), 15 cases being positive for lymph node metastasis, 40 cases being negtive for lymph node metastasis, 36 cases were stage Ⅰ, 4 cases were stage Ⅱ and 15 cases were stage Ⅲ. The median follow-up time was 20 (2-35) months. Delayed anastomotic leakage occurred in 1 patient and anastomotic stenosis occurred in 2 patients.Conclusion:Stent ileal bypass is safe and effective to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.

Objective To explore the policy foundations of early family education and rehabilitation support systems for children with developmental disabilities aged zero to three years,based on the International Classification of Functioning,Disability and Health(ICF),and to construct a comprehensive,whole-person and whole-lifecycle support system integrating early childhood education and rehabilitation services for families;and to propose corresponding sys-tem architecture,construction pathways and governance models.Methods Policy documents were compared,which were related to children's rehabilitation from the World Health Orga-nization,early education guidelines for children with disabilities from the United Nations Educational,Scientific and Cultural Organization,the U.S.Individuals with Disabilities Education Act,European Union children's reha-bilitation policies,and the"14th Five-Year Plan"from China Disabled Persons'Federation.The related policy frameworks and key content of early family education and rehabilitation were reviewed,and the composition,implementation pathways and governance models of the support system were systymatically analyzed.Results The support system consisted of early screening and assessment,family education and parent training,counsel-ing and personalized education and rehabilitation support,community support and resource integration,and re-mote digital support platforms.The study proposed the implementation pathways for five systems,including early screening and assessment,family education and parent training,counseling and personalized rehabilitation support,community support and resource integration,and remote digital support platforms.It emphasized gover-nance mechanisms such as multidisciplinary collaboration,interdepartmental coordination and support from re-mote digital platforms to build a continuous service chain from standardized assessments to interdepartmental collaboration.Conclusion The person-centered approach and whole life span development concept based on the ICF model,as well as the guiding principles of the health services continuum,provide systematic theoretical and policy support for early childhood education and rehabilitation for children with developmental disabilities aged zero to three years.Through multidisciplinary collaboration,interdepartmental coordination and the application of digital platforms,a scientific,continuous and child-centered support system can be built.This not only facilitates early detection and precise intervention but also promotes multi-party collaboration among families,communities and profes-sional institutions.It will further integrate disability prevention,rehabilitation and healthcare services,thereby im-proving children's functional abilities and family quality of life.

Objective To explore the policy foundations of early family education and rehabilitation support systems for children with developmental disabilities aged zero to three years,based on the International Classification of Functioning,Disability and Health(ICF),and to construct a comprehensive,whole-person and whole-lifecycle support system integrating early childhood education and rehabilitation services for families;and to propose corresponding sys-tem architecture,construction pathways and governance models.Methods Policy documents were compared,which were related to children's rehabilitation from the World Health Orga-nization,early education guidelines for children with disabilities from the United Nations Educational,Scientific and Cultural Organization,the U.S.Individuals with Disabilities Education Act,European Union children's reha-bilitation policies,and the"14th Five-Year Plan"from China Disabled Persons'Federation.The related policy frameworks and key content of early family education and rehabilitation were reviewed,and the composition,implementation pathways and governance models of the support system were systymatically analyzed.Results The support system consisted of early screening and assessment,family education and parent training,counsel-ing and personalized education and rehabilitation support,community support and resource integration,and re-mote digital support platforms.The study proposed the implementation pathways for five systems,including early screening and assessment,family education and parent training,counseling and personalized rehabilitation support,community support and resource integration,and remote digital support platforms.It emphasized gover-nance mechanisms such as multidisciplinary collaboration,interdepartmental coordination and support from re-mote digital platforms to build a continuous service chain from standardized assessments to interdepartmental collaboration.Conclusion The person-centered approach and whole life span development concept based on the ICF model,as well as the guiding principles of the health services continuum,provide systematic theoretical and policy support for early childhood education and rehabilitation for children with developmental disabilities aged zero to three years.Through multidisciplinary collaboration,interdepartmental coordination and the application of digital platforms,a scientific,continuous and child-centered support system can be built.This not only facilitates early detection and precise intervention but also promotes multi-party collaboration among families,communities and profes-sional institutions.It will further integrate disability prevention,rehabilitation and healthcare services,thereby im-proving children's functional abilities and family quality of life.

Objective To develop a reagent for detecting of 17α-hydroxyprogesterone(17α-OHP)in dried blood spots on filter paper fixed on vascular stent by Auto TRFIA-4 automatic fluorescence immunoanalyzer and evaluate its performance.Methods The microwell plate was coated with the sheep anti-rabbit IgG antibody as microwell reaction plate,the rabbit anti-human 17α-OHP antibody was diluted as intermediate antibody,and the 17α-OHP-bovine serum albumin conjugate was labeled by europium as europium marker.The concentration of 17α-OHP in dried blood spots on filter paper fixed on blood spots stent was quantitatively detected by Auto TRFIA-4 automatic fluorescence immunoanalyzer.The analysis sensitivity,accuracy,linearity,precision,specificity and stability were evaluated,and whether they met the requirements of the formulated industry standards were evaluated.A total of 227 neonatal heel blood filter paper samples from newborns who were born 72 hours after birth and within 7 days and fully breastfeeding were selected for reagent comparison test.The consistency was analyzed by χ2 test,Kappa test,t test,linear correlation analysis,regression analysis,Bland-Altman method analysis and predictive bias analysis of medical decision level,P<0.05 indicated statistically significant difference.Results The optimal coating concentration of sheep anti-rabbit IgG antibody was 3 μg/ml.The optimal dilution ratio of rabbit anti-human 17α-OHP antibody was 1∶1 500.The optimal dilution ratio of 17α-OHP europium marker mother liquor was 1∶2 500.The limit of blank,limit of detection,limit of quantification was 0.75,1.08 and 1.99 nmol/L,respectively.The relative deviations of the standard check test were within±15.00%,and the average recovery rate was 92.36%.The linear correlation coefficient was 0.997 1 in the range of 2.00 to 300.00nmo/L.The intra-assay and inter-assay coefficients of variation were all within 10.00%.The cross-reactivity rates of 100.00 ng/ml progesterone,17α-hydroxypreg nenolone and 11-deoxycortisol were within 0.089%to 0.64%.The performance of stability test met the requirements.The total coincidence rate was 100%compared with the results of contrast reagent.The quantitative results were highly correlated with the contrast reagent(r=0.999 4,tr=452.02,P<0.05).Conclusion The self-developed reagent has the advantages of high sensitivity,good accuracy,wide linear range,good precision,high specificity and good stability,which meets the requirements of the formulated industry standards,and has high correlation and consistency with the result of contrast reagent,which meets the needs of clinical detection.

Objective:To explore the safety and effectiveness of ileal bypass with stent to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.Methods:From Dec 2021 to Sep 2024, the clinical data of 55 patients undergoing robot anus-preserving surgery and stent ileal bypass for low rectal cancer in the First Affiliated Hospital of Ningbo University was analyzed retrospectively.Results:All the 55 patients successfully completed the operation. Among them, 47 patients underwent low anterior resection and 8 patients did sphincter resection. The total operation time was (207±37)min, the operation time of ileal bypass was (24±3)min, and the intraoperative blood loss was (32±18)ml. After operation, the time of stent disintegration was (23.93± 2.87) days, and the time of catheter removal was (29.55 ± 3.21) days. By postoperative TNM staging, there were 12 cases in T1 stage, 26 cases in T2 and 17 cases in T3. The number of lymph node dissection was (16.84±4.50), 15 cases being positive for lymph node metastasis, 40 cases being negtive for lymph node metastasis, 36 cases were stage Ⅰ, 4 cases were stage Ⅱ and 15 cases were stage Ⅲ. The median follow-up time was 20 (2-35) months. Delayed anastomotic leakage occurred in 1 patient and anastomotic stenosis occurred in 2 patients.Conclusion:Stent ileal bypass is safe and effective to prevent anastomotic leakage in Da Vinci robot anus-preserving surgery for low rectal cancer.

OBJECTIVE To study the pharmacological substance basis of Shaoyao gancao decoction for relieving acute and chronic pain. METHODS The antispasmodic effect of Shaoyao gancao decoction， ethyl acetate extract of Shaoyao gancao decoction and its effluent part of macroporous resin and 90% ethanol elution part of macroporous resin （the concentration of 4 drugs was 13.44 g/mL according to crude drug） was observed by in vitro small intestine tension test in rats. The acetic acid writhing test was conducted in mice to evaluate the analgesic effects of macroporous resin efflux site and macroporous resin 90% ethanol elution site （the dosage of 2.4 g/kg according to crude drug）. The levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL- 1β）， prostaglandin E2 （PGE2） and cyclooxygenase-2 （COX-2） in serum of mice were detected. The serum prototype and metabolites of mice after intragastric administration of macroporous resin 90% ethanol elution site were identified by high performance liquid chromatogre-time-of-flight mass spectrometry. RESULTS In vitro experiment showed that 90% ethanol eluting part of macroporous resin represented the best antispasmodic effect， and the inhibitory rate of small intestine tension was significantly higher than macroporous resin efflux site of Shaoyao gancao decoction （P＜0.05） without statistical significance， compared with Shaoyao gancao decoction （P＞0.05）. In the acetic acid writhing experiment， compared with model group， the writhing times of mice in the macroporous resin 90% ethanol elution part group were reduced significantly （P＜0.05）， the writhing latency was prolonged significantly （P＜0.05）， and the levels of COX-2， IL-1β， PGE2 and TNF-α in serum were decreased significantly （P＜0.05）. Ten kinds of protoproducts including paeoniflorin and glycyrrhizic acid were identified from serum of mice， and twenty-two kinds of metabolites including hydroxylated glycyrrhizin and glucosylated liquiritin were identified. CONCLUSIONS The effective part of Shaoyao gancao decoction for relieving acute and chronic pain is 90% ethanol elution part prepared by macroporous resin from the ethyl acetate extract. Ten components， including glycyrrhetinic acid and paeoniflorin， may be the basis of its pharmacological substances.