Objective: To investigate the spatio-temporal clustering characteristics of influenza in Ningxia Hui Autonomous Region from 2014 to 2023, so as to provide the basis for strengthening influenza prevention and control. Methods: Data pertaining to influenza cases reported in Ningxia Hui Autonomous Region from 2014 to 2023 were retrieved from the Infectious Disease Surveillance System of the Chinese Disease Prevention and Control Information System, including age, sex, current residence, onset date, and reporting date. The seasonal incidence of influenza was analyzed using seasonal index. The spatio-temporal clustering characteristics of influenza were identified using spatial autocorrelation analysis and spatio-temporal scan analysis. Results: A total of 20 377 influenza cases were reported in Ningxia Hui Autonomous Region from 2014 to 2023, with a male-to-female ratio of 1.15∶1. The majority were children under 15 years, with 10 950 cases accounting for 53.74%. Influenza was highly prevalent in January, February, March, and December, with seasonal indices of 219.06%, 111.00%, 246.65%, and 366.24%, respectively. The average annual reported incidence was 29.55/100 000, among which Pengyang County, Jinfeng District, Dawukou District, Xiji County, and Litong District had higher average annual reported incidence, at 63.99/100 000, 55.71/100 000, 55.70/100 000, 49.49/100 000, and 49.04/100 000, respectively. Spatial autocorrelation analysis showed that in 2023, there was spatial clustering of influenza cases in Ningxia Hui Autonomous Region (Moran's I=0.333, P<0.05), with a high-high cluster in Jingyuan County, while in other years, the distribution of influenza cases was random (all P>0.05). Spatio-temporal scan analysis showed that from 2014 to 2023, there were four space-time clusters in Ningxia Hui Autonomous Region, including one type Ⅰ cluster in Hongsibao District of Wuzhong City, with the clustering period from January 20 to 26, 2014; and three type Ⅱ clusters, mainly in January, February, March and December, covering one area in Shizuishan City, five areas in Guyuan City, one area in Zhongwei City, three areas in Wuzhong City, and four areas in Yinchuan City. Conclusions From 2014 to 2023, children under 15 years were the primary population affected by influenza in Ningxia Hui Autonomous Region, with distinct spatio-temporal distribution characteristics. The peak incidence occurred during the winter and spring seasons, and the main clustering areas were in the southern regions.

Objective:To discuss whether safranal affects the proliferation,migration,and phenotypic transformation of the vascular smooth muscle cells(VSMCs)in a high-glucose environment and to clarify the function of safranal in the prevention and treatment of diabetic(DM)vascular complications.Methods:The SD rats were selected as experimental subjects;primary VSMCs were cultured from rat thoracic aortas and divided into control group,25 mmol·L-1 high glucose(HG)group,HG+20 μmol·L-1 safranal group,HG+40 μmol·L-1 safranal group,and HG+80 μmol·L-1 safranal group.The cells in control group received no treatment;the cells in 25 mmol·L-1 HG group were pretreated with 25 mmol·L-1 HG;the cells in HG+20,40,and 80 μmol·L-1 safranal groups were further treated with 20,40,and 80 μmol·L-1 safranal respectively for 48 h on the basis of 25 mmol·L-1 HG group.Cell counting kit-8(CCK-8)method was used to determine the appropriate concentration of safranal and detect the viabilities of the VSMCs in various groups;cell scratch healing assay was used to detect the scratch healing rates of the VSMCs in various groups;Transwell chamber assay was used to detect the numbers of the migration VSMCs in various groups;immunofluorescence method was used to detect the fluorescence intensities of alpha-smooth muscle actin(α-SMA)and rabbit anti-osteopontin(OPN)in the VSMCs in various groups;Western blotting method was used to detect the expression levels of OPN,α-SMA,and proliferating cell nuclear antigen(PCNA)in the VSMCs in various groups.Results:Under microscope,on the 4th day of in vitro culture,the spindle-shaped or triangular cells crawled out from the edge of the thoracic aorta tissue blocks,with long spindle being the most common morphology.On the 14th,the cells gradually covered the bottom of the dish;when cell density reached 80%-90%,the characteristic"hills and valleys"growth pattern appeared.Third-generation cells were taken for immunofluorescence identification;immunofluorescence staining with VSMC-specific marker α-SMA showed positive expression of α-SMA protein in the primarily cultured VSMCs.The CCK-8 assay results showed that compared with control group,the cell viability of the cells in 160 μmol·L-1 safranal group was significantly decreased(P<0.01),indicating toxic damage to the cells.Under the conditions of safranal concentrations at 20,40,and 80 μmol·L-1 respectively,after 48 h intervention on VSMCs,no significant adverse effect on cell viability was observed;considering both the effect and toxicity of safranal,these three concentrations were used in subsequent cell experiments.After 48 h intervention,compared with control group,the activity of the VSMCs in 25 mmol·L-1 HG group was increased(P<0.001);compared with 25 mmol·L-1 HG group,the activities of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05).The cell scratch healing assay and Transwell assay results showed that after 48 h intervention,the scratch healing rate of the VSMCs in 25 mmol·L-1 HG group was significantly higher than that in control group(P<0.01),and the number of transmembrane cells through the Transwell chamber was significantly increased(P<0.05);compared with 25 mmol·L-1 HG group,the scratch healing rates of the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually decreased(P<0.05),and the number of transmembrane cells was decreased(P<0.05).The immunofluorescence staining results showed that compared with control group,the fluorescence intensity of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was significantly weakened(P<0.001),while the fluorescence intensity of OPN protein was significantly enhanced(P<0.001);compared with 25 mmol·L-1 HG group,the fluorescence intensities of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were gradually increased(P<0.05),and the fluorescence intensities of OPN were gradually weakened(P<0.05).The Western blotting method results showed that compared with control group,the expression level of α-SMA protein in the VSMCs in 25 mmol·L-1 HG group was decreased(P<0.05),and the expression levels of PCNA and OPN proteins were increased(P<0.01);compared with 25 mmol·L-1 HG group,the expression level of α-SMA protein in the VSMCs in HG+20,40,and 80 μmol·L-1 safranal groups were increased(P<0.05),and the expression levels of PCNA and OPN proteins were decreased(P<0.05).Conclusion:Safranal can inhibit the proliferation,migration,and phenotypic transformation of the VSMCs induced by high glucose.

Objective:To compare the efficacy of robot-assisted percutaneous reduction and screw fixation versus open reduction and plate fixation via the sinus tarsi approach in the treatment of Sanders types II and III calcaneal fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 82 patients (90 feet) with calcaneal fractures admitted to the Department of Orthopedic Trauma, Beijing Jishuitan Hospital, Capital Medical University from January 2020 to April 2024, including 74 males and 8 females, aged 24-87 years [(46.4±12.1)years]. According to Essex-Lopresti classification, the fractures were classified as tongue-type in 43 patients and joint-collapse-type in 47. According to Sanders classification, 69 feet were classified as type II and 21 as type III. Forty-seven patients (52 feet) were treated with robot-assisted percutaneous reduction and screw fixation (screw fixation group) and 35 (38 feet) with open reduction and plate fixation via the sinus tarsi approach (plate fixation group). The two groups were compared in terms of the operation duration, intraoperative blood loss, length of hospital stay and time to weight-bearing. The width, height, length, B?hler angle and Gissane angle of the calcaneus before surgery and at 1 day after surgery were compared. The Maryland foot and ankle function score, American Orthopedic Foot and Ankle Society (AOFAS) ankle and hindfoot function score, and visual analogue scale (VAS) score at 1, 3 months postoperatively, and at the last follow-up were compared. The incidence of postoperative complications and removal rate of internal fixation were also detected in the two groups.Results:All the patients were followed up for 9-60 months [(30.0±14.5)months]. There was no significant difference in the operation duration between the two groups ( P>0.05). The intraoperative blood loss, length of hospital stay and time to weight-bearing in the screw fixation group were 10.0(10.0, 20.0)ml, 7.0(5.0, 8.0)days and (5.0±0.8)weeks, which were significantly less or shorter than 30.0(20.0, 50.0)ml, 8.0(6.0, 11.0)days and (6.9±0.7)weeks in the plate fixation group ( P<0.05). The width, height, length, B?hler angle and Gissane angle of the calcaneus at 1 day after surgery were (43.4±4.2)mm, (46.2±4.0)mm, (81.6±5.1)mm, 27.1(20.4, 30.4)° and (113.4±10.1)° in the screw fixation group, which were all improved compared with those before surgery [(47.8±4.6)mm, (39.3±4.8)mm, (79.2±5.9)mm, 9.5(0.0,16.5)° and (119.3±13.4)°] ( P<0.01). The width, height, length and B?hler angle of the calcaneus at 1 day after surgery were (41.6±5.7)mm, (48.4±4.8)mm, (83.1±5.7)mm and 27.3(21.3, 31.6)° in the plate fixation group, which were all improved compared with those before surgery [(47.8±5.0)mm, (41.7±5.1)mm, (80.1±5.9)mm and 12.9(7.2,19.8)°] ( P<0.01), with no significant difference in the Gissane angle ( P>0.05). Before surgery and at 1 day postoperatively, no significant differences were found in the width, length, B?hler angle or Gissane angle of the calcaneus between the two groups ( P>0.05), while the height of the calcaneus in the screw fixation group was lower than that in the plate fixation group ( P<0.05). At 1 month after surgery and at the last follow-up, there were no significant differences in the Maryland foot and ankle function score, AOFAS ankle and hindfoot function score, and VAS score between the two groups ( P>0.05). At 3 months after surgery in the screw fixation group, the Maryland foot and ankle function score was (79.7±3.8)points, significantly higher than (74.7±2.8)points in the plate fixation group ( P<0.01); the AOFAS ankle and hindfoot function score was (77.1±5.0)points, significantly higher than (70.1±3.6)points in the plate fixation group ( P<0.01); the VAS score was 1.0(1.0, 2.0)points, significantly lower than 2.5(2.0, 3.0)points in the plate fixation group ( P<0.01). No significant difference was detected in the incidence of postoperative complications between the two groups ( P>0.05). The removal rate of internal fixation was 10% (5/52) in the screw fixation group, significantly lower than 29% (11/38) in the plate fixation group ( P<0.05). Conclusion:Compared with open reduction and plate fixation via the sinus tarsi approach, robot-assisted percutaneous reduction and screw fixation has the advantages of less intraoperative blood loss, shorter hospital stay, earlier weight-bearing exercises, better early functional recovery and pain relief, and lower internal fixation removal rate in the treatment of Sanders types II and III calcaneal fractures.

Objective:To analyze the trabecular bone score (TBS) and its association with muscle health in the postmenopausal population.Methods:It was a cross-sectional study. A total of 214 postmenopausal individuals who underwent dual-energy X-ray absorptiometry testing at the Beijing Research Institute of Sports Science between January and December 2023 were consecutively included. The participants were divided into two groups based on the presence or absence of osteoporosis (OP): 62 cases in the OP group and 152 cases in the non-OP group. All subjects completed body composition assessments and scans of bilateral hip and lumbar spine bone mineral density (BMD)(All the BMD in this study were areal BMD). Demographic data, including age, age at menarche, age at menopause, and fracture history, were also collected. TBS was calculated using the TBS iNsight software. Muscle health indicators included upper limb, lower limb, trunk, and total muscle mass, lean body mass, appendicular skeletal muscle mass index (ASMI), and grip strength. The Pearson correlation analysis was used to determine the relationship between TBS and muscle health indicators, as well as between TBS and the BMD of lumbar spine (L 1-4). Further, multiple linear regression analysis was conducted to explore the independent association between TBS and muscle health indicators. Results:TBS, lean body mass, ASMI, grip strength, and muscle mass of the upper limb, lower limb, trunk, whole body in the OP group were all lower than those in the non-OP group [(1.27±0.07) vs (1.35±0.07) score, (36.64±3.45) vs (39.14±3.62) kg, (6.16±0.56) vs (6.44±0.63) kg/m2, (23.87±3.31) vs (25.34±4.33) kg, (3.54±0.47) vs (3.78±0.47) kg, (11.70±1.33) vs (12.68±1.49) kg, (16.57±1.68) vs (17.50±1.69) kg, and (34.91±3.33) vs (37.13±3.47) kg] (all P<0.05). Pearson analysis showed that TBS was positively correlated with lean body mass, ASMI, grip strength, and muscle mass of the upper limb, lower limb, whole body (all P<0.05). TBS was negatively correlated with age ( P<0.01). TBS was positively correlated with the BMD of lumbar spine (L 1-4) ( r=0.660, P<0.01). Multiple regression analysis demonstrated that ASMI was positively correlated with TBS ( β=0.284, P<0.01). Conclusion:In the postmenopausal population, individuals with OP have lower TBS. Furthermore, TBS is closely associated with muscle health.

Objective To investigate the mechanisms by which astaxanthin(AST)alleviates oxaliplatin(OXA)-induced neuropathic pain through antioxidant pathways so as to provide theoretical basis for clinical intervention.Methods Animal experiments:SD rats were divided into five groups(n=6):control group,OXA(4 mg/kg)group,OXA+Oil group,OXA+AST(5 mg/kg)group,and OXA+AST(10 mg/kg)group.Mechanical and cold pain thresholds were measured at day 0,7,14,and 21.Malondialdehyde(MDA)content and superoxide dismutase(SOD)activity in the dorsal root ganglia(DRG)were detected using the thiobarbituric acid(TBA)method and WST-1 assay,respectively.Western blotting was performed to analyze the expressions of Nrf2 and HO-1.Cell experiments:neuro-2a cells were divided into control group,OXA(50 μmol/L)group,AST(10 μmol/L)group,and OXA(50 μmol/L)+AST(10 μmol/L)group.Cells were treated with nerve growth factor(NGF,50 ng/mL)to induce growth,and morphological changes were observed under an inverted microscope.Intracellular reactive oxygen species(ROS)level and mitochondrial superoxide were measured using DCFH-DA fluorescent probe and MitoSOXTM red,respectively.Mitochondrial function was assessed by JC-1 assay.Western blotting was used to detect Nrf2 and HO-1 expressions.Results Animal experiments:① Mechanical and cold pain thresholds were reduced in OXA and OXA+Oil groups(P＜0.05),while AST significantly increased these thresholds in OXA-treated rats(P＜0.05).② SOD activity decreased while MDA content increased in the DRG of OXA-treated rats(P＜0.05).AST restored SOD activity and reduced MDA level(P＜0.05,P＜0.01).③ Western blotting showed elevated Nrf2 and HO-1 expressions in OXA group(P＞0.05),which were further upregulated by AST(P＜0.05,P＜0.01).Cell experiments:① OXA reduced the number of neurite-bearing cells and shortened the average neurite length(P＜0.05).Inverted microscopic observation revealed that AST intervention increased both parameters(P＜0.01,P＜0.001).② OXA increased intracellular and mitochondrial ROS fluorescence intensity(P＜0.05),which was attenuated by AST(P＜0.01).③ JC-1 assay revealed decreased mitochondrial membrane potential in OXA group(P＜0.01),which was partially reversed by AST(P＜0.05).④ Western blotting results showed that OXA upregulated Nrf2 and HO-1 expressions(P＜0.05,P＜0.01),and AST further enhanced their levels(P＜0.01).Conclusion AST alleviates OXA-induced neuropathic pain by promoting Nrf2/HO-1 expression,enhancing SOD activity,reducing lipid peroxidation and ROS production,and improving mitochondrial function.

Objective To investigate the mechanisms by which astaxanthin(AST)alleviates oxaliplatin(OXA)-induced neuropathic pain through antioxidant pathways so as to provide theoretical basis for clinical intervention.Methods Animal experiments:SD rats were divided into five groups(n=6):control group,OXA(4 mg/kg)group,OXA+Oil group,OXA+AST(5 mg/kg)group,and OXA+AST(10 mg/kg)group.Mechanical and cold pain thresholds were measured at day 0,7,14,and 21.Malondialdehyde(MDA)content and superoxide dismutase(SOD)activity in the dorsal root ganglia(DRG)were detected using the thiobarbituric acid(TBA)method and WST-1 assay,respectively.Western blotting was performed to analyze the expressions of Nrf2 and HO-1.Cell experiments:neuro-2a cells were divided into control group,OXA(50 μmol/L)group,AST(10 μmol/L)group,and OXA(50 μmol/L)+AST(10 μmol/L)group.Cells were treated with nerve growth factor(NGF,50 ng/mL)to induce growth,and morphological changes were observed under an inverted microscope.Intracellular reactive oxygen species(ROS)level and mitochondrial superoxide were measured using DCFH-DA fluorescent probe and MitoSOXTM red,respectively.Mitochondrial function was assessed by JC-1 assay.Western blotting was used to detect Nrf2 and HO-1 expressions.Results Animal experiments:① Mechanical and cold pain thresholds were reduced in OXA and OXA+Oil groups(P＜0.05),while AST significantly increased these thresholds in OXA-treated rats(P＜0.05).② SOD activity decreased while MDA content increased in the DRG of OXA-treated rats(P＜0.05).AST restored SOD activity and reduced MDA level(P＜0.05,P＜0.01).③ Western blotting showed elevated Nrf2 and HO-1 expressions in OXA group(P＞0.05),which were further upregulated by AST(P＜0.05,P＜0.01).Cell experiments:① OXA reduced the number of neurite-bearing cells and shortened the average neurite length(P＜0.05).Inverted microscopic observation revealed that AST intervention increased both parameters(P＜0.01,P＜0.001).② OXA increased intracellular and mitochondrial ROS fluorescence intensity(P＜0.05),which was attenuated by AST(P＜0.01).③ JC-1 assay revealed decreased mitochondrial membrane potential in OXA group(P＜0.01),which was partially reversed by AST(P＜0.05).④ Western blotting results showed that OXA upregulated Nrf2 and HO-1 expressions(P＜0.05,P＜0.01),and AST further enhanced their levels(P＜0.01).Conclusion AST alleviates OXA-induced neuropathic pain by promoting Nrf2/HO-1 expression,enhancing SOD activity,reducing lipid peroxidation and ROS production,and improving mitochondrial function.

Objective:To compare the efficacy of robot-assisted percutaneous reduction and screw fixation versus open reduction and plate fixation via the sinus tarsi approach in the treatment of Sanders types II and III calcaneal fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 82 patients (90 feet) with calcaneal fractures admitted to the Department of Orthopedic Trauma, Beijing Jishuitan Hospital, Capital Medical University from January 2020 to April 2024, including 74 males and 8 females, aged 24-87 years [(46.4±12.1)years]. According to Essex-Lopresti classification, the fractures were classified as tongue-type in 43 patients and joint-collapse-type in 47. According to Sanders classification, 69 feet were classified as type II and 21 as type III. Forty-seven patients (52 feet) were treated with robot-assisted percutaneous reduction and screw fixation (screw fixation group) and 35 (38 feet) with open reduction and plate fixation via the sinus tarsi approach (plate fixation group). The two groups were compared in terms of the operation duration, intraoperative blood loss, length of hospital stay and time to weight-bearing. The width, height, length, B?hler angle and Gissane angle of the calcaneus before surgery and at 1 day after surgery were compared. The Maryland foot and ankle function score, American Orthopedic Foot and Ankle Society (AOFAS) ankle and hindfoot function score, and visual analogue scale (VAS) score at 1, 3 months postoperatively, and at the last follow-up were compared. The incidence of postoperative complications and removal rate of internal fixation were also detected in the two groups.Results:All the patients were followed up for 9-60 months [(30.0±14.5)months]. There was no significant difference in the operation duration between the two groups ( P>0.05). The intraoperative blood loss, length of hospital stay and time to weight-bearing in the screw fixation group were 10.0(10.0, 20.0)ml, 7.0(5.0, 8.0)days and (5.0±0.8)weeks, which were significantly less or shorter than 30.0(20.0, 50.0)ml, 8.0(6.0, 11.0)days and (6.9±0.7)weeks in the plate fixation group ( P<0.05). The width, height, length, B?hler angle and Gissane angle of the calcaneus at 1 day after surgery were (43.4±4.2)mm, (46.2±4.0)mm, (81.6±5.1)mm, 27.1(20.4, 30.4)° and (113.4±10.1)° in the screw fixation group, which were all improved compared with those before surgery [(47.8±4.6)mm, (39.3±4.8)mm, (79.2±5.9)mm, 9.5(0.0,16.5)° and (119.3±13.4)°] ( P<0.01). The width, height, length and B?hler angle of the calcaneus at 1 day after surgery were (41.6±5.7)mm, (48.4±4.8)mm, (83.1±5.7)mm and 27.3(21.3, 31.6)° in the plate fixation group, which were all improved compared with those before surgery [(47.8±5.0)mm, (41.7±5.1)mm, (80.1±5.9)mm and 12.9(7.2,19.8)°] ( P<0.01), with no significant difference in the Gissane angle ( P>0.05). Before surgery and at 1 day postoperatively, no significant differences were found in the width, length, B?hler angle or Gissane angle of the calcaneus between the two groups ( P>0.05), while the height of the calcaneus in the screw fixation group was lower than that in the plate fixation group ( P<0.05). At 1 month after surgery and at the last follow-up, there were no significant differences in the Maryland foot and ankle function score, AOFAS ankle and hindfoot function score, and VAS score between the two groups ( P>0.05). At 3 months after surgery in the screw fixation group, the Maryland foot and ankle function score was (79.7±3.8)points, significantly higher than (74.7±2.8)points in the plate fixation group ( P<0.01); the AOFAS ankle and hindfoot function score was (77.1±5.0)points, significantly higher than (70.1±3.6)points in the plate fixation group ( P<0.01); the VAS score was 1.0(1.0, 2.0)points, significantly lower than 2.5(2.0, 3.0)points in the plate fixation group ( P<0.01). No significant difference was detected in the incidence of postoperative complications between the two groups ( P>0.05). The removal rate of internal fixation was 10% (5/52) in the screw fixation group, significantly lower than 29% (11/38) in the plate fixation group ( P<0.05). Conclusion:Compared with open reduction and plate fixation via the sinus tarsi approach, robot-assisted percutaneous reduction and screw fixation has the advantages of less intraoperative blood loss, shorter hospital stay, earlier weight-bearing exercises, better early functional recovery and pain relief, and lower internal fixation removal rate in the treatment of Sanders types II and III calcaneal fractures.

Objective:To analyze the trabecular bone score (TBS) and its association with muscle health in the postmenopausal population.Methods:It was a cross-sectional study. A total of 214 postmenopausal individuals who underwent dual-energy X-ray absorptiometry testing at the Beijing Research Institute of Sports Science between January and December 2023 were consecutively included. The participants were divided into two groups based on the presence or absence of osteoporosis (OP): 62 cases in the OP group and 152 cases in the non-OP group. All subjects completed body composition assessments and scans of bilateral hip and lumbar spine bone mineral density (BMD)(All the BMD in this study were areal BMD). Demographic data, including age, age at menarche, age at menopause, and fracture history, were also collected. TBS was calculated using the TBS iNsight software. Muscle health indicators included upper limb, lower limb, trunk, and total muscle mass, lean body mass, appendicular skeletal muscle mass index (ASMI), and grip strength. The Pearson correlation analysis was used to determine the relationship between TBS and muscle health indicators, as well as between TBS and the BMD of lumbar spine (L 1-4). Further, multiple linear regression analysis was conducted to explore the independent association between TBS and muscle health indicators. Results:TBS, lean body mass, ASMI, grip strength, and muscle mass of the upper limb, lower limb, trunk, whole body in the OP group were all lower than those in the non-OP group [(1.27±0.07) vs (1.35±0.07) score, (36.64±3.45) vs (39.14±3.62) kg, (6.16±0.56) vs (6.44±0.63) kg/m2, (23.87±3.31) vs (25.34±4.33) kg, (3.54±0.47) vs (3.78±0.47) kg, (11.70±1.33) vs (12.68±1.49) kg, (16.57±1.68) vs (17.50±1.69) kg, and (34.91±3.33) vs (37.13±3.47) kg] (all P<0.05). Pearson analysis showed that TBS was positively correlated with lean body mass, ASMI, grip strength, and muscle mass of the upper limb, lower limb, whole body (all P<0.05). TBS was negatively correlated with age ( P<0.01). TBS was positively correlated with the BMD of lumbar spine (L 1-4) ( r=0.660, P<0.01). Multiple regression analysis demonstrated that ASMI was positively correlated with TBS ( β=0.284, P<0.01). Conclusion:In the postmenopausal population, individuals with OP have lower TBS. Furthermore, TBS is closely associated with muscle health.

In order to establish a stable in vitro culture platform for chicken small intestine three-dimensional (3D) organoids, in this study, crypt cells were collected from the small intestine of 18-day-old embryos of AA broilers. On the basis of the L-WRN conditioned medium, we optimized the culture conditions of chicken small intestinal organoids by adjusting the proportions of nicotinamide, N-acetylcysteine, LY2157299, CHIR99021, Jagged-1, FGF, and other cytokines to select the medium suitable for the long-term stable growth of the organoids. The optimization results showed that the addition of 1.5 µmol/L CHIR99021 significantly improved the organoid formation efficiency and organoid diameter. When 0.5 µmol/L Jagged-1 was added, a small amount of bud-like tissue appeared in organoids. After the addition of 50 ng/mL FGF-2, the rate of organoid germination was significantly increased. The 1.5 µmol/L CHIR99021, 0.5 µmol/L Jagged-1, and 50 ng/mL FGF-2 added in the medium can cooperate with each other to improve the formation and speed up the proliferation and differentiation of organoids, while improving the stemness maintenance of cells. The morphology, cell types, and culture characteristics of chicken small intestinal organoids were studied by HE staining, transmission electron microscopy, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR), indirect immunofluorescence, and immunohistochemistry. The results showed that the 3D organoids of the chicken small intestine cultured in vitro were morphologically consistent with the chicken intestinal tissue and contained differentiated epithelial cells. In summary, we successfully established an in vitro culture system for chicken small intestinal organoids, providing a new method for the subsequent research on chicken intestinal physiology, pathology, and host-pathogen interaction mechanism and the development of relevant drugs.
Animals ; Organoids/metabolism* ; Intestine, Small/drug effects* ; Chickens ; Cell Culture Techniques/methods* ; Culture Media ; Chick Embryo ; Tissue Culture Techniques/methods*