1.Current quality status and management countermeasures of occupational health technical services in Zhejiang Province
Qiuliang XU ; Feng HAN ; Peng WANG ; Zhen ZHOU ; Fei LI ; Hongwei XIE ; Yong HU ; Weiming YUAN ; Lifang ZHOU ; Hua ZOU
Journal of Environmental and Occupational Medicine 2026;43(3):341-346
Background The quality of occupational health technical services is directly linked to the protection of workers' health rights and the efficacy of occupational disease prevention and control. However, the industry still faces critical challenges: sporadic instances of institutional non-compliance and persistent irregularities in professional practice continue to undermine overall service performance. Objective To assess the current quality status of occupational health technical services in Zhejiang Province and propose countermeasures for quality improvement, providing a scientific basis for policy optimization and service delivery quality enhancement. Methods A total of 69 occupational health technical service institutions in Zhejiang Province that obtained formal accreditation as of April 30, 2024, were sampled, including 3 public institutions and 66 private institutions (comprising 3 formerly Class-A, 28 formerly Class-B, 11 formerly Class-C, and 24 newly certified institutions). Following the Technical Protocol for Quality Monitoring of Occupational Health Technical Service in Zhejiang Province and the Technical Protocol for Proficiency Testing of Occupational Health Detection in Zhejiang Province, a quality assessment task force comprising national and provincial experts was established. Evaluation was conducted across four dimensions: qualification maintenance and compliance, standardization of technical services, authenticity of technical services, and proficiency testing, utilizing a combination of document review, on-site inspections, and technical skill assessments. Results The occupational health technical service institutions in Zhejiang Province were predominantly private entities (82.5%), with significant disparities in overall service quality. The pass rates for qualification maintenance and compliance, technical service standardization, technical service authenticity, and the excellence rate for laboratory proficiency testing were 81.5%, 80.7%, 97.3%, and 90.4%, respectively. Regarding qualification maintenance, the pass rates for "environmental conditions" (49.8%, 56.7%) and "instrumentation and equipment" (58.2%、65.6%) were significantly lower for formerly Class-C and newly certified institutions compared to other categories. In terms of technical standardization, "standardized on-site inspections" recorded the lowest pass rate (67.4%), with newly certified institutions at only 48.0%. Regarding technical service authenticity, formerly Class-C institutions exhibited issues such as missing raw chromatograms for blank samples (85.7% pass rate). In laboratory proficiency testing, public and formerly Class-A institutions achieved 100% excellence rates, but the performance of formerly Class-C and newly certified institutions was comparatively weak; specifically, the failure rate for organic analysis in formerly Class-C institutions reached 20%; the failure rate for dust testing items in newly certified institutions was 10.3%. Conclusion The overall quality of occupational health technical services in Zhejiang Province still requires significant improvement, particularly in basic institutional conditions, the standardization of on-site inspections, and laboratory proficiency in organic and dust analysis. Formerly Class-C and newly certified institutions should be the primary focus of quality management efforts. Differentiated regulatory strategies are recommended, alongside strengthening interim and ex-post supervision to gradually enhance the quality of occupational health technical services across all institutions.
2.Risk Factors Analysis and Predictive Model Construction for Acute Kidney Injury Following Amphotericin B Deoxycholate Use in Hospitalized Patients
Hao XIE ; Yixun SHI ; Zhiqing XU ; Minquan LI ; Xiaoli DU ; Gang CHEN ; Bin ZHAO
Medical Journal of Peking Union Medical College Hospital 2026;17(2):429-437
To investigate the risk factors for acute kidney injury (AKI) following the use of amphotericin B deoxycholate and to develop a predictive model to guide clinical monitoring and intervention. A retrospective analysis was conducted on hospitalized patients who received amphotericin B deoxycholate between January 2014 and September 2024. Patients were divided into a training set and a validation set. Demographic data, laboratory findings, and medication orders were collected. Based on the occurrence of AKI during treatment and within 7 days after discontinuation, patients were classified into an AKI group and a non-AKI group. Univariate analysis was used to screen for potential risk factors, multivariate logistic regression was employed to construct a predictive model, and model performance was evaluated using the area under the receiver operating characteristic curve (AUC) and the Hosmer-Lemeshow test. The training set included 473 patients, comprising 255 males (53.91%) and 218 females (46.09%), with a median age of 52(35, 62) years. The AKI group consisted of 191 cases (40.38%), and the non-AKI group consisted of 282 cases (59.62%). The validation set included 114 patients, comprising 80 males (70.18%) and 34 females (29.82%), with a median age of 43.5 (31.0, 58.5) years. The AKI group consisted of 42 cases (36.84%), and the non-AKI group consisted of 72 cases (63.16%). Univariate analysis revealed statistically significant differences between the two groups in 23 factors (all Admission to the ICU, elevated serum creatinine at admission, and comorbid cardiac insufficiency as potential risk factors for AKI, while prophylactic use of diphenhydramine/promethazine or sodium bicarbonate showed a protective association. A predictive model with good discrimina-tion and calibration was developed, which may provide a basis for early identification of high-risk patients and timely adjustment of treatment strategies in clinical practice.
3.Mechanism of modified Lichong decoction on apoptosis of transplanted tumor cells in nude mice via Fas/FasL pathway and caspase apoptosis pathway
Guoshan SHI ; Tiexia HAO ; Zhe CHEN ; Xu LI ; Liwei MA
China Pharmacy 2026;37(7):895-901
OBJECTIVE To investigate the effect and mechanism of modified Lichong decoction (MLCD) on the apoptosis of transplanted tumor cells in nude mice. METHODS Human gastric cancer AGS cells were cultured, and a nude mice transplanted tumor model was established. The nude mice were divided into the model group and MLCD low-, medium- and high-dose groups (150, 300, 600 mg/kg). They were given distilled water or the corresponding drug solution by gavage once daily for four consecutive weeks. The size of transplanted tumors in nude mice was measured every six days, and the tumor volume was calculated. After the medication, the nude mice were sacrificed, and the transplanted tumor tissues were isolated. The contents of lactate dehydrogenase (LDH) and reactive oxygen species (ROS) in the transplanted tumor tissues were detected, and the changes in mitochondrial membrane potential were assessed. The pathological morphological changes were observed. The enzymatic activities of caspase-3, caspase-8, and caspase-9, as well as protein expressions of Fas and FasL and mRNA expressions of caspase-3, caspase-8, caspase-9, Fas and FasL in the transplanted tumor tissues, were detected. RESULTS Compared with the model group, the volume of transplanted tumors in nude mice from all MLCD dose groups was reduced to varying degrees. The contents of LDH and ROS, as well as the enzymatic activities of caspase-3, caspase-8 and caspase-9, were significantly increased/enhanced. The mitochondrial membrane potential was significantly decreased. The protein expressions of Fas and FasL, and the mRNA expressions of caspase-3, caspase-8, caspase-9, Fas and FasL were significantly up-regulated. Most of these differences were statistically significant ( P <0.05 or P <0.01). Pathological results showed that with increasing doses of MLCD, the cellular density in the transplanted tumor tissues gradually decreased, and typical morphological features of apoptosis, such as loosening and increasing fragmentation, became more prominent. CONCLUSIONS MLCD can induce apoptosis in transplanted tumor cells of nude mice, and its mechanism may be related to the activation of the Fas/FasL pathway and the caspase apoptotic pathway.
4.In Vitro Anti-psoriatic Effect of Kangfuxin Liquid via Inhibiting Cell Proliferation and Migration Ability and Blocking JAK3/STAT3 Signaling Pathway
Shuai LI ; Xuan LIU ; Wenyan TANG ; Zhenqi WU ; Chunhui CHEN ; Dadan QIU ; Yi XU ; Chenggui ZHANG ; Jianquan ZHU ; Jiali ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):123-133
ObjectiveThis paper aims to explore the in vitro anti-psoriasis activity and potential mechanism of Kangfuxin liquid (KFX liquid), providing experimental evidence for the anti-psoriasis effect of KFX liquid. MethodsFirstly, the uninduced human immortalized keratinocyte cells (HaCaT cells) were divided into seven groups, namely the control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). After being treated with different concentrations of KFX liquid, the effect of KFX liquid on the normal cell proliferation was detected by using the cell counting kit-8 (CCK-8) method. Secondly, the uninduced HaCaT cells were divided into six groups, namely the control group and recombinant human interleukin-7A (rh-IL-7A) groups with different doses (5, 10, 50, 100, 120 g·L-1). After being treated with different concentrations of recombinant human interleukin-17A (rh IL-17A) liquid, the effect of rh IL-17A on cell proliferation was detected. The optimal induction concentration was screened. Then, normal HaCaT cells were divided into a control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). Except for the control group, the other groups established psoriasis cell models with the optimal induction concentration of rh IL-17A. After being treated with different concentrations of KFX liquid, the effects of KFX liquid on the psoriasis-like HaCaT cell proliferation were investigated. Finally, the uninduced HaCaT cells were divided into six groups, namely the control group, rh IL-17A group, methotrexate (MTX) group, and KFX liquid groups with different doses (20, 40, 80 g·L-1). Except for the control group, the other groups used the optimal induction concentration of rh IL-17A to establish psoriasis cell models. After being treated with different drugs, the cell migration levels were detected through scratch assays, and real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the relative mRNA expression levels of Ki-67 antigen (Ki67), S100 calcium-binding protein A7 (S100A7), S100 calcium-binding protein A8 (S100A8), and S100 calcium-binding protein A9 (S100A9), thereby comprehensively evaluating the in vitro anti-psoriasis activity of KFX liquid. By detecting the relative mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and chemokine-20 (CXCL-20) inflammatory-related factors in psoriasis-like HaCaT cells and the protein expression levels of Janus kinase 3 (JAK3), phosphorylated Janus kinase 3 (p-JAK3), signal transducer and activator of transcription 3 (STAT3), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3), the mechanism was explored. ResultsCompared with that of control group, when treated with 80 g·L-1 KFX liquid for 72 h (P<0.05) and at different times with 160 g·L-1 KFX liquid, the HaCaT cell proliferation activity was significantly affected (P<0.01), while the other concentrations of KFX liquid had no significant differences in cell morphology and cell proliferation activity at different times, indicating that the KFX liquid is relatively safe for HaCaT cells and has no obvious toxic side effects. Compared with that of control group, when treated with different concentrations of rh IL-17A for 24 h, the HaCaT cell proliferation activity was significantly enhanced, and the cell activity was the strongest when the concentration was 100 μg·L-1 (P<0.05), with a density close to 100% and intact cell morphology, indicating that 100 μg·L-1 is the optimal concentration for inducing HaCaT cell proliferation. The results of the KFX liquid treatment on rh IL-17A-induced psoriasis-like cells show that the KFX liquid not only effectively inhibits the rh IL-17A-induced psoriasis-like HaCaT cell proliferation activity (P<0.01), but also significantly reduces the migration ability of rh IL-17A-induced psoriasis-like HaCaT cells (P<0.01), and the relative mRNA expression levels of Ki67, S100A7, S100A8, and S100A9 (P<0.01). Moreover, the KFX liquid can significantly reduce the relative mRNA expression levels of IL-1β, IL-6, and CXCL-20 in rh IL-17A-induced psoriasis-like cells (P<0.01), and significantly inhibit the phosphorylation levels of JAK3 and STAT3 proteins (P<0.05, P<0.01). ConclusionThe KFX liquid has no obvious toxicity to uninduced HaCaT cells. It can inhibit rh IL-17A-induced psoriasis-like HaCaT cell proliferation, reduce the cell migration ability, and has good in vitro anti-psoriasis activity. Its action mechanism may be related to downregulating the expression levels of inflammation-related cytokines in the JAK3/STAT3 signaling pathway and inhibiting the phosphorylation levels of JAK3 and STAT3 proteins.
5.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
ObjectiveTo optimize the extraction method for polysaccharides from turnip(Brassica rapa), and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction(BP-1) and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index, a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio, extraction time and extraction temperature, in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry(UV), gas chromatography-mass spectrometry(GC-MS), Congo red assay, and Fourier-transform infrared spectroscopy(FT-IR) to determine purity, monosaccharide composition, triple-helix structure, and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy(SEM) and atomic force microscopy(AFM). Zebrafish were divided into the blank group(adding E3 medium), and BP-1-1, BP-1-10, BP-1-50, BP-1-200, BP-1-1 000 groups(adding BP-1 solutions at concentrations of 1, 10, 50, 200, 1 000 mg·L-1, respectively), and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype, survival rate, malformation rate, and heart rate. Experimental animals were randomly divided into the blank group, model group, BP-1-10 group(10 mg·L-1), BP-1-50 group(50 mg·L-1), and BP-1-200 group(200 mg·L-1). The blank group was cultured in E3 medium, the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization(dpf) zebrafish embryos using 10 g·L-1 of glucose combined with 500 µmol·L-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution, while the blank and model groups received an equal volume of saline. Glucose and insulin(INS) levels were measured using enzyme-linked immunosorbent assay(ELISA) kits, the effects on the liver were observed by hematoxylin-eosin(HE) histopathological sections. The mRNA expression levels of glucagon(Glucagon), insulin(Insa), and phosphoenolpyruvate carboxykinase 1(PCK1) were detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). ResultsThe optimized extraction conditions were determined as follows:solid-to-liquid ratio of 1∶40(g·mL-1), extraction time of 66 min, and extraction temperature of 79 ℃. Under these conditions, the yield of turnip polysaccharides was (10.34±0.96)%. UV analysis indicated that BP-1 contained no proteins or nucleic acids, GC-MS analysis revealed that BP-1 consisted of six monosaccharides(arabinose, rhamnose, ribose, mannose, galactose and glucose). Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure, FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids, SEM analysis revealed an irregular flaky structure with a flat and smooth surface, AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L-1. Pharmacodynamic results showed that, compared with the blank group, the model group exhibited significantly increased glucose levels and significantly decreased INS levels(P<0.01). Compared with the model group, the BP-1-50 group significantly reduced glucose levels and increased INS levels(P<0.05). Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal, with hepatocytes appearing plump. Real-time PCR results showed that, compared with the blank group, the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1, and significantly downregulated mRNA expression of Insa(P<0.01). Compared with the model group, the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1, and significantly upregulated mRNA expression of Insa(P<0.01). ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate, is simple to operate, has low costs, making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides, contains α-glycosidic bonds and uronic acids, exhibits hypoglycemic activity, and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.
6.Traditional Chinese Medicine Regulation of Wnt/β-catenin Signaling Pathway for Post-stroke Cognitive Impairment Treatment: A Review
Wanyue XU ; Yanjie LI ; Haoyuan LIU ; Bohua WAN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):337-345
Post-stroke cognitive impairment (PSCI) is a common and severe complication in stroke patients, significantly affecting their quality of life and social function. Despite increasing research on PSCI in recent years, effective therapeutic methods remain limited. The Wnt/β-catenin signaling pathway has emerged as a critical research focus in neuroscience due to its essential role in neuroprotection, neurorepair, and cognitive recovery. Dysregulation of the Wnt/β-catenin pathway is considered one of the key mechanisms in the onset and progression of PSCI. Traditional Chinese medicine (TCM), with its multi-component, multi-target, and synergistic properties, has shown unique advantages in modulating the Wnt/β-catenin signaling pathway, providing a potential novel approach for PSCI treatment. TCM regulates the Wnt/β-catenin pathway through various mechanisms and exerts effects such as inhibiting cell apoptosis, maintaining blood-brain barrier integrity, reducing neuroinflammation, promoting neuroplasticity, and enhancing neurorepair, thereby improving post-stroke cognitive function. This review summarized the latest research progress on the regulation of the Wnt/β-catenin signaling pathway by TCM in intervening PSCI. It analyzed the mechanisms of action of various TCM components and compound formulas within this pathway, aiming to provide a theoretical basis for innovative strategies for PSCI treatment in the future and offer new research insights and practical guidance for the application of TCM in cerebrovascular diseases.
7.Related genes, pathogenesis, and lncRNA functions in retinoblastoma
Qihang DIAO ; Shuangxiu LI ; Xinyan XU ; Rongyu GAO ; Mengjun FU
International Eye Science 2026;26(5):816-822
Retinoblastoma(RB)represents the most common primary intraocular malignant tumor in infants and young children, posing a severe threat to the visual acuity and life of affected patients. Clinically, it is categorized into hereditary and non-hereditary subtypes. Mounting evidence indicates that RB cells most likely originate from cone photoreceptor precursor cells, and the tumorigenesis is closely associated with the inactivation of the RB1 gene. Beyond RB1, a growing list of genes including MYCN, TP53 and PRMT1 have been implicated in the initiation and progression of RB. Concurrently, the dysregulation of multiple signaling pathways such as RB/E2F, WNT, and PI3K/AKT synergistically drives the survival, proliferation, invasion, and metastasis of RB tumor cells. The therapeutic paradigm for RB has undergone a dramatic shift from the conventional enucleation-dominated approach to personalized multimodal therapies that prioritize globe salvage and visual preservation, encompassing local therapies, chemotherapy and radiotherapy. Moreover, novel therapeutic modalities including targeted therapy, immunotherapy and gene therapy are currently under active preclinical and clinical investigation. In recent years, long non-coding RNAs(lncRNAs), as pivotal regulators of genetic expression, have attracted increasing attention for their critical roles in RB oncogenesis and progression. These molecules hold great promise to serve as novel diagnostic biomarkers and offer innovative insights and strategies for RB treatment. This review summarizes the latest research advances in the aforementioned aspects of retinoblastoma.
8.The effects of tetrandrine tablets on inflammatory cytokines and acute phase response following the first-dose of zoledronic acid in postmenopausal osteoporosis patients
Zhanfei XU ; Li LI ; Xiangyang TONG ; Xuechao ZHAO
Journal of Pharmaceutical Practice and Service 2026;44(4):216-220
Objective To investigate the effect of tetrandrine tablets on inflammatory cytokines and acute phase response following the first-dose of zoledronic acid in postmenopausal osteoporosis patients. Methods 80 postmenopausal osteoporosis patients receiving the first-dose of zoledronic acid admitted in the Affliated Xuzhou Municipal Hospital of Xuzhou Medical University from June, 2022 to December, 2024 were selected as study objects, and the patients were randomly divided into control group (40 cases, prophylactic treatment with acetaminophen tablets) and study group (40 cases, prophylactic treatment with tetrandrine tablets and acetaminophen tablets). The high-sensitivity C-reactive protein (hs-CRP), interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α (TNF-α) and the occurrence of acute phase reactions (fever, muscle and joint pain, gastrointestinal discomfort) between two groups were compared. Results The levels of serum hs-CRP, IL-1β, IL-6, and TNF-α in both groups significantly increased on day 1 and day 3 after infusion of zoledronic acid (all P values <0.05). The levels of serum hs-CRP, IL-6, and TNF-α in the control group were higher than those in the study group on day 1 and 3 after infusion of zoledronic acid, with statistical significance (all P values <0.05). In terms of acute phase reactions, the incidence of fever, muscle and joint pain in the control group after infusion of zoledronic acid were higher than those in the study group, with statistical significance (P<0.05). No statistically significant difference in the incidence of gastrointestinal discomfort between the two groups (P>0.05). Conclusion Tetrandrine tablets could reduce the expression levels of hs-CRP, IL-1β, IL-6, and TNF-α following the first-dose of zoledronic acid in postmenopausal osteoporosis patients, reduce the incidence of acute phase reactions, and alleviate adverse reactions.
9.In Vitro Anti-psoriatic Effect of Kangfuxin Liquid via Inhibiting Cell Proliferation and Migration Ability and Blocking JAK3/STAT3 Signaling Pathway
Shuai LI ; Xuan LIU ; Wenyan TANG ; Zhenqi WU ; Chunhui CHEN ; Dadan QIU ; Yi XU ; Chenggui ZHANG ; Jianquan ZHU ; Jiali ZHU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):123-133
ObjectiveThis paper aims to explore the in vitro anti-psoriasis activity and potential mechanism of Kangfuxin liquid (KFX liquid), providing experimental evidence for the anti-psoriasis effect of KFX liquid. MethodsFirstly, the uninduced human immortalized keratinocyte cells (HaCaT cells) were divided into seven groups, namely the control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). After being treated with different concentrations of KFX liquid, the effect of KFX liquid on the normal cell proliferation was detected by using the cell counting kit-8 (CCK-8) method. Secondly, the uninduced HaCaT cells were divided into six groups, namely the control group and recombinant human interleukin-7A (rh-IL-7A) groups with different doses (5, 10, 50, 100, 120 g·L-1). After being treated with different concentrations of recombinant human interleukin-17A (rh IL-17A) liquid, the effect of rh IL-17A on cell proliferation was detected. The optimal induction concentration was screened. Then, normal HaCaT cells were divided into a control group and KFX liquid groups with different doses (5, 10, 20, 40, 80, 160 g·L-1). Except for the control group, the other groups established psoriasis cell models with the optimal induction concentration of rh IL-17A. After being treated with different concentrations of KFX liquid, the effects of KFX liquid on the psoriasis-like HaCaT cell proliferation were investigated. Finally, the uninduced HaCaT cells were divided into six groups, namely the control group, rh IL-17A group, methotrexate (MTX) group, and KFX liquid groups with different doses (20, 40, 80 g·L-1). Except for the control group, the other groups used the optimal induction concentration of rh IL-17A to establish psoriasis cell models. After being treated with different drugs, the cell migration levels were detected through scratch assays, and real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the relative mRNA expression levels of Ki-67 antigen (Ki67), S100 calcium-binding protein A7 (S100A7), S100 calcium-binding protein A8 (S100A8), and S100 calcium-binding protein A9 (S100A9), thereby comprehensively evaluating the in vitro anti-psoriasis activity of KFX liquid. By detecting the relative mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), and chemokine-20 (CXCL-20) inflammatory-related factors in psoriasis-like HaCaT cells and the protein expression levels of Janus kinase 3 (JAK3), phosphorylated Janus kinase 3 (p-JAK3), signal transducer and activator of transcription 3 (STAT3), and phosphorylated signal transducer and activator of transcription 3 (p-STAT3), the mechanism was explored. ResultsCompared with that of control group, when treated with 80 g·L-1 KFX liquid for 72 h (P<0.05) and at different times with 160 g·L-1 KFX liquid, the HaCaT cell proliferation activity was significantly affected (P<0.01), while the other concentrations of KFX liquid had no significant differences in cell morphology and cell proliferation activity at different times, indicating that the KFX liquid is relatively safe for HaCaT cells and has no obvious toxic side effects. Compared with that of control group, when treated with different concentrations of rh IL-17A for 24 h, the HaCaT cell proliferation activity was significantly enhanced, and the cell activity was the strongest when the concentration was 100 μg·L-1 (P<0.05), with a density close to 100% and intact cell morphology, indicating that 100 μg·L-1 is the optimal concentration for inducing HaCaT cell proliferation. The results of the KFX liquid treatment on rh IL-17A-induced psoriasis-like cells show that the KFX liquid not only effectively inhibits the rh IL-17A-induced psoriasis-like HaCaT cell proliferation activity (P<0.01), but also significantly reduces the migration ability of rh IL-17A-induced psoriasis-like HaCaT cells (P<0.01), and the relative mRNA expression levels of Ki67, S100A7, S100A8, and S100A9 (P<0.01). Moreover, the KFX liquid can significantly reduce the relative mRNA expression levels of IL-1β, IL-6, and CXCL-20 in rh IL-17A-induced psoriasis-like cells (P<0.01), and significantly inhibit the phosphorylation levels of JAK3 and STAT3 proteins (P<0.05, P<0.01). ConclusionThe KFX liquid has no obvious toxicity to uninduced HaCaT cells. It can inhibit rh IL-17A-induced psoriasis-like HaCaT cell proliferation, reduce the cell migration ability, and has good in vitro anti-psoriasis activity. Its action mechanism may be related to downregulating the expression levels of inflammation-related cytokines in the JAK3/STAT3 signaling pathway and inhibiting the phosphorylation levels of JAK3 and STAT3 proteins.
10.Extraction,Separation and Hypoglycemic Activity Analysis of Polysaccharides from Brassica rapa
Mengyu HOU ; Ruina XU ; Qingsong LI ; Shaoxuan LI ; Xinying MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(10):219-228
ObjectiveTo optimize the extraction method for polysaccharides from turnip(Brassica rapa), and analyze and evaluate the primary structure of the isolated and purified turnip polysaccharide fraction(BP-1) and its hypoglycemic effects in diabetic zebrafish. MethodsTaking polysaccharide yield as the evaluation index, a semi-bionic extraction method was employed. Single-factor experiments and Box-Behnken response surface methodology were used to investigate three factors of solid-to-liquid ratio, extraction time and extraction temperature, in order to optimize the extraction process. BP-1 was isolated and purified using the Sevage method and DEAE-52 cellulose column chromatography. Structural characterization of the turnip polysaccharides was performed using ultraviolet-visible spectrophotometry(UV), gas chromatography-mass spectrometry(GC-MS), Congo red assay, and Fourier-transform infrared spectroscopy(FT-IR) to determine purity, monosaccharide composition, triple-helix structure, and functional groups. The microstructure of the polysaccharides was observed using scanning electron microscopy(SEM) and atomic force microscopy(AFM). Zebrafish were divided into the blank group(adding E3 medium), and BP-1-1, BP-1-10, BP-1-50, BP-1-200, BP-1-1 000 groups(adding BP-1 solutions at concentrations of 1, 10, 50, 200, 1 000 mg·L-1, respectively), and zebrafish embryos were subjected to a 96-hour exposure experiment. The maximum tolerated concentration of BP-1 in zebrafish was determined by evaluating its effects on phenotype, survival rate, malformation rate, and heart rate. Experimental animals were randomly divided into the blank group, model group, BP-1-10 group(10 mg·L-1), BP-1-50 group(50 mg·L-1), and BP-1-200 group(200 mg·L-1). The blank group was cultured in E3 medium, the model and treatment groups were induced to establish a diabetic model in 4 day-post-fertilization(dpf) zebrafish embryos using 10 g·L-1 of glucose combined with 500 µmol·L-1 of alloxan. The treatment groups received corresponding doses of BP-1 solution, while the blank and model groups received an equal volume of saline. Glucose and insulin(INS) levels were measured using enzyme-linked immunosorbent assay(ELISA) kits, the effects on the liver were observed by hematoxylin-eosin(HE) histopathological sections. The mRNA expression levels of glucagon(Glucagon), insulin(Insa), and phosphoenolpyruvate carboxykinase 1(PCK1) were detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). ResultsThe optimized extraction conditions were determined as follows:solid-to-liquid ratio of 1∶40(g·mL-1), extraction time of 66 min, and extraction temperature of 79 ℃. Under these conditions, the yield of turnip polysaccharides was (10.34±0.96)%. UV analysis indicated that BP-1 contained no proteins or nucleic acids, GC-MS analysis revealed that BP-1 consisted of six monosaccharides(arabinose, rhamnose, ribose, mannose, galactose and glucose). Congo red assay indicated that the molecular conformation did not exhibit a triple-helix structure, FT-IR analysis showed the presence of α-glycosidic bonds and uronic acids, SEM analysis revealed an irregular flaky structure with a flat and smooth surface, AFM analysis suggested that the aggregated structure might be formed by the entanglement of molecular chains and intramolecular hydrogen bonding. The maximum tolerated concentration of BP-1 in zebrafish over 96 h was determined to be 200 mg·L-1. Pharmacodynamic results showed that, compared with the blank group, the model group exhibited significantly increased glucose levels and significantly decreased INS levels(P<0.01). Compared with the model group, the BP-1-50 group significantly reduced glucose levels and increased INS levels(P<0.05). Histopathological examination of liver tissue revealed that various doses of BP-1 had a certain reparative effect on damaged liver tissue. The liver tissue structure in the BP-1-200 group was nearly normal, with hepatocytes appearing plump. Real-time PCR results showed that, compared with the blank group, the model group exhibited significantly upregulated mRNA expressions of Glucagon and PCK1, and significantly downregulated mRNA expression of Insa(P<0.01). Compared with the model group, the BP-1-50 and BP-1-200 groups showed significantly downregulated mRNA expressions of Glucagon and PCK1, and significantly upregulated mRNA expression of Insa(P<0.01). ConclusionThe semi-bionic extraction method for turnip polysaccharides yields a high extraction rate, is simple to operate, has low costs, making it suitable for large-scale industrial production. BP-1 consists of six monosaccharides, contains α-glycosidic bonds and uronic acids, exhibits hypoglycemic activity, and provides a certain protective effect on the liver of alloxan-induced diabetic model zebrafish.

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