Objective To investigate the effects and mechanisms of fatty acid binding protein 4(FABP4)on ferroptosis in human renal tubular epithelial cells(HK-2)treated with hypoxia/reoxygenation(H/R).Methods HK-2 cells were cultured in vitro and subjected to hypoxia for 24 hours followed by reoxygenation for different durations(1,3,6 h).The messenger RNA(mRNA)and protein levels of FABP4 in HK-2 cells were detected at each time point using real-time fluorescent quantitative polymerase chain reaction and Western blotting.Small interfering RNA(siRNA)technology was used to silence the expression of FABP4 gene in HK-2 cells,which were then treated with H/R(24 h of hypoxia and 6 h of reoxygenation)or treated with the Nrf2 inhibitor ML385.Cell proliferation activity was assessed using cell counting kit-8(CCK-8).Lactate dehydrogenase(LDH)levels were measured by enzyme-linked immune absorbent assay.Malondialdehyde(MDA),glutathione(GSH)and ferrous ion(Fe2+)levels were determined by biochemical technology.Reactive oxygen species(ROS)levels were detected using the 2',7'-dichlorodihydrofluorescein diacetate fluorescence probe.Protein expression levels of FABP4,nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),glutathione peroxidase 4(GPX4)and solute carrier family 7 member 11(SLC7A11)were measured by Western blotting.Results The mRNA and protein levels of FABP4 in HK-2 cells increased with prolonged reoxygenation time(all P<0.05).H/R treatment reduced cell proliferation activity,increased LDH levels in the cell supernatant,and elevated MDA,Fe2+and ROS levels in HK-2 cells while decreasing GSH levels and the protein levels of Nrf2,HO-1,GPX4 and SLC7A11(all P<0.05).Silencing FABP4 enhanced the proliferation activity of H/R-treated HK-2 cells(P<0.05),reduced MDA,Fe2+and ROS levels,increased GSH levels,and elevated the protein levels of Nrf2,HO-1,GPX4 and SLC7A11(all P<0.05).However,these beneficial effects of FABP4 silencing on H/R-induced ferroptosis in HK-2 cells were reversed by co-treatment with ML385.Conclusions Silencing FABP4 alleviated H/R-induced ferroptosis in HK-2 cells,possibly by activating the Nrf2/GPX4 axis.

Objective:To investigate the positive rate,mutation type and distribution characteristics of thalassemia gene detection in children in Nanchong area.Methods:The common α and β-thalassemia gene mutation sites were detected in 1 254 children suspected of thalassemia by hematological screening in our hospital from January 2017 to December 2023,and the genotypes,detection rates and clinical characteristics of thalassemia in local children were statistically analyzed.Results:Among 1 254 children with suspected thalassemia,490 carriers were screened out,with a positive detection rate of 39.07％.Among them,220 cases(17.54％)were α-thalassemia,251 cases(20.02％)wereβ-thalassemia,and 19 cases(1.52％)were αβ compound thalassemia.Among 220 cases of α-thalassemia,the main genotypes were--SEA/αα,-α3.7/αα,-α3.7/--SEA and-α4.2/αα,accounting for 63.64％,18.64％,5.91％,and 5.00％,respectively.Among 251 cases of β-thalassemia,CD17,CD41-42,and IVS-Ⅱ-654 genotypes were the most common,accounting for 40.24％,29.88％,and 17.93％,respectively.In 19 cases of αβ compound thalassemia,the most common genotypes were-α3.7/αα compound CD41-42,--SEA/αα compound CD41-42,--SEA/αα compound CD17,accounting for 26.32％,15.79％,and 15.79％,respectively.In addition,compared with healthy individuals,red blood cell(RBC)in the thalassemia gene carriers was significantly increased,while hemoglobin(Hb),mean corpuscular volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration(MCHC)and red blood cell distribution width-standard deviation(RDW-SD)were significantly decreased(all P＜0.01).The ROC curve analysis showed that the area under the curve(AUC)of RDW-SD,MCHC,MCH,MCV,Hb and RBC were 0.827,0.707,0.823,0.863,0.603 and 0.882,respectively.The thalassemia gene carrying rates from 2017 to 2023 in Nanchong were 35.6％(54/154),28.43％(56/197),34.74％(74/213),40.56％(58/143),42.69％(73/171),45.86％(83/181),and 47.18％(92/195),respectively,showing an upward trend year by year.Conclusion:The positive detection rate of children's thalassemia gene in Nanchong is relatively high,and the genetic types are complex,with β-thalassemia as the main type.The genetic pattern shows obvious regional distribution characteristics.The genotypes of thalassemia in children are mainly--SEA/αα,-α3.7/αα,CD17,CD41-42 and IVS-Ⅱ-654,which are consistent with the genotypes of adults in this area,but different from high-risk areas such as Dongguan and Guangxi.

Objective:To investigate the positive rate,mutation type and distribution characteristics of thalassemia gene detection in children in Nanchong area.Methods:The common α and β-thalassemia gene mutation sites were detected in 1 254 children suspected of thalassemia by hematological screening in our hospital from January 2017 to December 2023,and the genotypes,detection rates and clinical characteristics of thalassemia in local children were statistically analyzed.Results:Among 1 254 children with suspected thalassemia,490 carriers were screened out,with a positive detection rate of 39.07％.Among them,220 cases(17.54％)were α-thalassemia,251 cases(20.02％)wereβ-thalassemia,and 19 cases(1.52％)were αβ compound thalassemia.Among 220 cases of α-thalassemia,the main genotypes were--SEA/αα,-α3.7/αα,-α3.7/--SEA and-α4.2/αα,accounting for 63.64％,18.64％,5.91％,and 5.00％,respectively.Among 251 cases of β-thalassemia,CD17,CD41-42,and IVS-Ⅱ-654 genotypes were the most common,accounting for 40.24％,29.88％,and 17.93％,respectively.In 19 cases of αβ compound thalassemia,the most common genotypes were-α3.7/αα compound CD41-42,--SEA/αα compound CD41-42,--SEA/αα compound CD17,accounting for 26.32％,15.79％,and 15.79％,respectively.In addition,compared with healthy individuals,red blood cell(RBC)in the thalassemia gene carriers was significantly increased,while hemoglobin(Hb),mean corpuscular volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration(MCHC)and red blood cell distribution width-standard deviation(RDW-SD)were significantly decreased(all P＜0.01).The ROC curve analysis showed that the area under the curve(AUC)of RDW-SD,MCHC,MCH,MCV,Hb and RBC were 0.827,0.707,0.823,0.863,0.603 and 0.882,respectively.The thalassemia gene carrying rates from 2017 to 2023 in Nanchong were 35.6％(54/154),28.43％(56/197),34.74％(74/213),40.56％(58/143),42.69％(73/171),45.86％(83/181),and 47.18％(92/195),respectively,showing an upward trend year by year.Conclusion:The positive detection rate of children's thalassemia gene in Nanchong is relatively high,and the genetic types are complex,with β-thalassemia as the main type.The genetic pattern shows obvious regional distribution characteristics.The genotypes of thalassemia in children are mainly--SEA/αα,-α3.7/αα,CD17,CD41-42 and IVS-Ⅱ-654,which are consistent with the genotypes of adults in this area,but different from high-risk areas such as Dongguan and Guangxi.

Objective:To study the effect of angiopoietin-1(Ang-1)and tyrosine kinase receptor 2(Tie2)inhibitor on glucose transportation in the human umbilical vein endothelial cells(HUVECs)cultured under high glucose conditions,and to clarify its mechanism.Methods:The HUVECs were cultured in high glucose(30 mmol·L?1)in vitro and treated with 0,200,500,1 000,and 2 000 μg·L?1 Ang-1 and 0,2 500,5 000,and 7 500 nmol·L?1 Tie2 inhibitor;cell counting kit-8(CCK-8)method was used to detect the cell activity to screen the optimal concentrations of Ang-1 and Tie2 inhibitor.Glucose kit was used to detect the glucose level in the supernatant of the HUVECs after Ang-1 intervention.The HUVECs were randomly divided into blank control group(NG group),high glucose group(HG group),HG+Tie2 inhibitor group(HG+In-Tie2 group),HG+Ang-1 group,HG+Ang-1+Tie2 inhibitor group(HG+Ang-1+In-Tie2 group),and HG+Ang-1+phosphatidylinositol 3-kinase(PI3K)inhibitor group(HG+Ang-1+LY294002 group).5-Ethynyl-2'-deoxyuridine(EdU)method was used to detect the proliferation activities of the cells in various groups;YO-PRO-1/PI method was used to detect the apoptotic rates of the cells in various groups;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Ang-1 mRNA and Tie2 mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of Tie2,glucose transporter 1(GLUT1),and glucose transporter 4(GLUT4)proteins and the ratios of phosphorylated PI3K(p-PI3K)/PI3K and phosphorylated protein kinase B(p-AKT)/AKT in the cells in various groups.Results:The CCK-8 assay results showed that compared with 0 μg·L?1 Ang-1 group,the activity of the HUVECs was significantly increased after treated with 200 μg·L?1 Ang-1 for 48 h(P<0.01);compared with 0 nmol·L?1 Tie2 inhibitor group,the activity of the HUVECs was significantly decreased after treated with 2 500、5 000 and 7 500 nmol·L?1 Tie2 inhibitor(P<0.01);the optimal concentrations of Ang-1 and Tie2 inhibitor were 200 μg·L?1 and 2 500 nmol·L?1,respectively.Compared with NG group,the glucose level in the supernatant of the HUVECs in HG group was significantly increased(P<0.01);compared with HG group,the glucose level in the supernatant of the HUVECs in Ang-1 group was significantly decreased(P<0.01).The EdU assay results showed that compared with NG group,the proliferation activity of the HUVECs in HG group was significantly decreased(P<0.01);compared with HG group,the proliferation activity of the HUVECs in HG+In-Tie2 group was significantly decreased(P<0.01),and the proliferation activity of the HUVECs in HG+Ang-1 group was significantly increased(P<0.01);compared with HG+Ang-1 group,the proliferation activities of the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased(P<0.01).The YO-PRO-1/PI assay results showed that compared with NG group,the apoptotic rate of the HUVECs in HG group was significantly increased(P<0.01);compared with HG group,the apoptotic rate of the HUVECs in HG+In-Tie2 group was significantly increased(P<0.01),and the apoptotic rate of the HUVECs in HG+Ang-1 group was significantly decreased(P<0.01);compared with HG+Ang-1 group,the apoptotic rates of the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly increased(P<0.01).The RT-qPCR results showed that compared with NG group,the expression levels of Ang-1 mRNA and Tie2 mRNA in the HUVECs in HG group and HG+In-Tie2 group were significantly decreased(P<0.01);compared with HG group,the expression levels of Ang-1 mRNA and Tie2 mRNA in HG+In-Tie2 group were significantly decreased(P<0.01),and the expression levels of Ang-1 mRNA and Tie2 mRNA in the HUVECs in HG+Ang-1 group were significantly increased(P<0.05);compared with HG+Ang-1 group,the expression levels of Ang-1 mRNA and Tie2 mRNA in the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased(P<0.05 or P<0.01).The Western blotting results showed that compared with NG group,the expression level of Tie2 protein in the HUVECs in HG group was significantly decreased(P<0.01),and the expression levels of GLUT1 and GLUT4 proteins were significantly increased(P<0.01);compared with HG group,the expression levels of Tie2,GLUT1,and GLUT4 proteins in the HUVECs in HG+In-Tie2 group were significantly decreased(P<0.01),the expression level of Tie2 protein in the HUVECs in HG+Ang-1 group was significantly increased(P<0.01),and the expression levels of GLUT1 and GLUT4 proteins were significantly decreased(P<0.01);compared with HG+Ang-1 group,the expression levels of Tie2,GLUT1,and GLUT4 proteins in the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased(P<0.01).Compared with NG group,the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG group were significantly increased(P<0.01);compared with HG group,the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG+In-Tie2 group were significantly decreased(P<0.01),and the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG+Ang-1 group were significantly decreased(P<0.01);compared with HG+Ang-1 group,the p-PI3K/PI3K and p-AKT/AKT ratios in the HUVECs in HG+Ang-1+In-Tie2 group and HG+Ang-1+LY294002 group were significantly decreased(P<0.01).Conclusion:Ang-1 down-regulates the expressions of GLUT1 and GLUT4 in the HUVECs cultured under high glucose conditions;the binding of Ang-1 to Tie2 may down-regulate GLUT1 and GLUT4 via the PI3K/AKT signaling pathway to participate in the glucose transportation in the HUVECs cultured under high glucose conditions.

OBJECTIVE: To explore clinical efficacy of autologous platelet-rich plasma(PRP) in treating Rockwood type Ⅲ acromioclavicular dislocation. METHODS: From January 2019 to July 2021, 32 patients with Rockwood type Ⅲ acromioclavicular dislocation were treated with minimally invasive adjustable titanium plate internal fixation, and were divided into PRP group and control group according to whether PRP treatment was performed, with 16 patients in each group. In PRP group, there were 10 males and 6 females, aged from 28 to 47 years old with an average of (36.75±7.14) years old;the time from injury to surgery ranged from 1 to 31 h with an average of (26.13±3.98) h;5 patients on the left side and 11 patients on the right side;PRP was injected once during operation and the 4th and 8th weeks after operation respectively. In control group, there were 8 males and 8 females, aged from 30 to 52 years old with an average of (38.50±5.48) years old; the time from injury to surgery ranged from 1 to 29 h with an average of (25.48±3.11) h;7 patients on the left side and 9 patients on the right side; minimally invasive surgical treatment was performed. Visual analogue scale(VAS) was used to evaluate pain and Constant-Murley score for shoulder joint function was used to evaluate the recovery of shoulder joint movement function before operation and 1, 3, 6, and 12 months after operation respectively. RESULTS: All patients were followed up for 12 to 28 months with an average of (18.3±5.2) months. All incisions patients healed well without adverse events such as infection. Postoperative VAS of PRP group at 1, 3, and 6 months were (5.5±1.2), (3.7±1.6), and (2.4±1.2), respectively, while were lower than those of control group (6.6±1.4), (4.9±1.1), and (3.7±1.3), respectively;and had statistical differences between two groups (P<0.05). There was no statistically significant difference in VAS between two groups before operation and 12 months after operation (P>0.05). Postoperative Constant-Murley scores of PRP group at 1, 3, and 6 months were (64.09±11.61), (73.19±12.89), and (82.61±14.81) points, respectively, which were higher than those of control group were (52.32±17.42), (61.65±14.43), and (72.52±11.04) respectively;and the differences were statistically significant (P<0.05). There was no statistically significant difference in Constant-Murley scores at 12 months after operation between two groups (P>0.05). In PRP group, there was no statistically significant difference at 6 months and 12 months after operation (P>0.05), while there were statistically significant differences at the other time points (1 month after operation compared with before operation, 3 months after operation compared with 6 months after operation, and 3 months after operation compared with 1 month after operation) (P<0.05). In control group, there was no statistically significant difference when comparing 1 month and 3 months after operation (P>0.05), while at the other time points (1 month after operation with before operation, 3 months after operation with 6 months after operation, and 6 months after operation with 12 months after operation), the differences were all statistically significant (P<0.05). CONCLUSION Adjustable titanium plate fixation combined with postoperative injection of PRP for the treatment of Rockwood type III acromioclavicular joint dislocation has effect of promoting the recovery of shoulder joint function and reducing pain.
Humans ; Male ; Female ; Adult ; Middle Aged ; Platelet-Rich Plasma ; Acromioclavicular Joint/surgery* ; Bone Plates ; Titanium ; Joint Dislocations/therapy* ; Fracture Fixation, Internal/methods*

Objective To investigate the effects and mechanisms of fatty acid binding protein 4(FABP4)on ferroptosis in human renal tubular epithelial cells(HK-2)treated with hypoxia/reoxygenation(H/R).Methods HK-2 cells were cultured in vitro and subjected to hypoxia for 24 hours followed by reoxygenation for different durations(1,3,6 h).The messenger RNA(mRNA)and protein levels of FABP4 in HK-2 cells were detected at each time point using real-time fluorescent quantitative polymerase chain reaction and Western blotting.Small interfering RNA(siRNA)technology was used to silence the expression of FABP4 gene in HK-2 cells,which were then treated with H/R(24 h of hypoxia and 6 h of reoxygenation)or treated with the Nrf2 inhibitor ML385.Cell proliferation activity was assessed using cell counting kit-8(CCK-8).Lactate dehydrogenase(LDH)levels were measured by enzyme-linked immune absorbent assay.Malondialdehyde(MDA),glutathione(GSH)and ferrous ion(Fe2+)levels were determined by biochemical technology.Reactive oxygen species(ROS)levels were detected using the 2',7'-dichlorodihydrofluorescein diacetate fluorescence probe.Protein expression levels of FABP4,nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HO-1),glutathione peroxidase 4(GPX4)and solute carrier family 7 member 11(SLC7A11)were measured by Western blotting.Results The mRNA and protein levels of FABP4 in HK-2 cells increased with prolonged reoxygenation time(all P<0.05).H/R treatment reduced cell proliferation activity,increased LDH levels in the cell supernatant,and elevated MDA,Fe2+and ROS levels in HK-2 cells while decreasing GSH levels and the protein levels of Nrf2,HO-1,GPX4 and SLC7A11(all P<0.05).Silencing FABP4 enhanced the proliferation activity of H/R-treated HK-2 cells(P<0.05),reduced MDA,Fe2+and ROS levels,increased GSH levels,and elevated the protein levels of Nrf2,HO-1,GPX4 and SLC7A11(all P<0.05).However,these beneficial effects of FABP4 silencing on H/R-induced ferroptosis in HK-2 cells were reversed by co-treatment with ML385.Conclusions Silencing FABP4 alleviated H/R-induced ferroptosis in HK-2 cells,possibly by activating the Nrf2/GPX4 axis.

Objective:To investigate the clinical value immature granulocyte percentage(IG％)and other inflammatory indicators in the severity of acute appendicitis.Methods:A total of 201 pediatric patients undergoing appendicitis surgery admitted to Zhoukou Central Hospital from June 2022 to August 2023 were included.Patients with pathologically confirmed actue appendici-tis were divided two subgroups:actue simple appendicitis(ASA)group and actue complicated ap-pendicitis(ACA)group,The variables that included IG％,white blood cell(WBC)count,absolute neutrophil count(ANC),absolute lymphocyte count(ALC),neutrophil to lymphocyte ratio(NLR),pro-calcitonin(PCT),C-reactive protein(CRP),platelet to lymphocyte(PLR)and other indexes were ana-lyzed between ASA and ACA group.The logistic regression model for diagnosis of ACA was es-tablished,and the diagostic value of this model and other inflammtory indicators for ACA was evaluated by receiver operating characteristic(ROC)curve analysis.Results:The levels of IG％,WBC,ANC,ALC,NLR,PCT and PLR were higher and the level of ALC was lower in ACA group than those in ASA group(all P＜0.05).Logistic regression analysis showed that IG％,NLR and CRP were three diagnostic determinants of ACA(all P＜0.05).The AUC of the established logistic model and IG％,NLR,CRP were 0.868,0.821,0.691 and 0.790(all P＜0.001).The logistic model was vali-dated by independent cohorts,and the AUC was 0.872,the sensitivity was 90.0％and the speci-ficity was 75.6％.Conlusions:The IG％value can early indicator for pediatric ACA,and the es-tablished logistic regression model based on biomarkers including IG％,NLR and CRP has clinical value in diagnosing ACA in children.

Objective:To screen and diagnose cervical lesions in middle-aged and elderly postmenopausal women and analyze the characteristics of cervical lesions in middle-aged and elderly women.Methods:A retrospective analysis was conducted on the screening data of 1 337 postmenopausal women aged between 50 and 86 years who visited the outpatient clinics of the gynecology department of Beijing Hospital between January, 2015 and December, 2019.Participants were divided into groups based on age, with 787 in the 50-59 years old group, 483 in the 60-69 years old group, 60 in the 70-79 years old group, and 7 in the 80-89 years old group.Exfoliated cervical cells were collected, and high-risk human papillomavirus(HPV)detection and liquid-based cervical cytology examination were performed to confirm the diagnosis based on the pathological results of cervical biopsy.Cervical lesions were analyzed.Results:53.0%(708)of the 1 337 enrolled patients made their hospital visits because of symptoms.There were 666 patients(49.8%)with cervical lesions, including 357 with low-grade squamous intraepithelial lesions(LSIL), 252 with high-grade squamous intraepithelial lesions(HSIL)and 57(4.3%)with cervical cancer.The 70-79 group showed the highest incidence of HSIL at 30.0%(18/60), while the 50~59 group had the highest incidence of LSIL at 30.6%(241/787), and there were significant differences in different types of cervical lesions between different age groups( χ2=28.919, P<0.01). Liquid-based cervical cytology examination revealed a positive rate of 51.6%(166/322)for LSIL, a positive rate of 70.8%(143/202)for HSIL, and a rate of 71.4%(25/35)for cervical cancer.There were significant differences in LSIL, HSIL and other above-mentioned positive rates of lesions( P<0.05). In the cervical malignancy group, cytological high-grade squamous epithelial lesions made up 51.4%(18/35). Intraepithelial lesions or cancerous lesions represented 29.2%(59/202)in the HSIL group.The rate of intraepithelial lesions or cancerous lesions in the LSIL group was 48.4%(156/322), with statistically significance in these differences( χ2=118.806, P<0.01). Chronic cervicitis was found in 52.9%(109)of 206 patients with atypical squamous cells and positive HPV, and in 76.2%(16)of 21 patients without HPV.HPV examination was completed in 1 301 patients, with 69.4%(903)having infections by one HPV strain and 30.6%(398)having infections by mixed HPV strains.Biopsy results showed that the rates of HPV infections in LSIL, HSIL and cancer patients were 84.6%(302/357), 95.9(185/193)and 100%(54/54), respectively, and the differences in these pathological results were statistically significant( P<0.01). HPV16 was the most common subtype in single HPV strain infections.The most common subtypes of HPV infections in cervical lesions with HSIL and above were HPV16 and HPV18.The agreement rate was 52.4%(187/357)for LSIL, 60.7%(153/252)for HSIL, and 64.9%(37/57)for cervical cancer between diagnoses based on cervical biopsy and those based on colposcopy.As the severity of lesions increased, the accuracy of colposcopy also increased.Differences between colposcopic diagnosis and pathological diagnosis on different types of cervical lesions were statistically significant( χ2=550.382, P<0.01).1 310 patients underwent endocervical curettage, identifying 120 cases of HSIL and 39 cases of cervical cancer, with a detection rate of 55.8%(159/285). Conclusions:The incidence of cervical cancer is high in elderly women.The incidences of high-grade lesions and cervical cancer in patients with atypical squamous cells plus HPV infections in the cervix are also high.Postmenopausal patients with cervical lesions are mainly infected by a single HPV subtype, with subtype 16 being the most common one.With increasing severity of lesions, the accuracy of colposcopy-based diagnosis increases and the rate of missed diagnosis decreases.Cervical curettage is an important means to reduce missed diagnosis with colposcopy.

Excessive sodium/salt intake is the leading dietary risk factor for the loss of healthy life in the Chinese population. The "Healthy China 2030" Action Plan set the goal of reducing salt intake by 20% by 2030. However, salt intake in China is still at a very high level in the world, with adults reaching 11 g/d, more than twice the recommended limit of 5 g/d. The current policies and action plans of China have targeted catering workers, children, adolescents, and home chefs in salt, oil, and sugar reduction actions. However, there are still obvious deficiencies in the coordinated promotion and implementation. This study, therefore, proposed a set of comprehensive strategies (named CHRPS that is composed of communication and education, salt reduction in home cooking, salt reduction in restaurants, reducing salt content in pre-packaged food, and surveillance and evaluation) and key implementation points for further deepening the salt reduction action in China. These strategies were developed based on the main sources of dietary sodium for Chinese residents, the status of "knowledge, attitude and practice" in salt reduction, evidence of effective intervention measures, existing policies and requirements, and the salt reduction strategies of the World Health Organization and experience from some other countries. As a scientific reference, the CHRPS strategies will help the government and relevant organizations quickly implement salt reduction work and facilitate the earlier realization of China's salt reduction goal.
Adult ; Child ; Adolescent ; Humans ; Sodium Chloride, Dietary ; Sodium, Dietary ; Diet ; Food ; China

OBJECTIVES: To explore the cytotoxicity of four wild mushrooms involved in a case of Yunnan sudden unexplained death (YNSUD), to provide the experimental basis for prevention and treatment of YNSUD. METHODS: Four kinds of wild mushrooms that were eaten by family members in this YNSUD incident were collected and identified by expert identification and gene sequencing. Raw extracts from four wild mushrooms were extracted by ultrasonic extraction to intervene HEK293 cells, and the mushrooms with obvious cytotoxicity were screened by Cell Counting Kit-8 (CCK-8). The selected wild mushrooms were prepared into three kinds of extracts, which were raw, boiled, and boiled followed by enzymolysis. HEK293 cells were intervened with these three extracts at different concentrations. The cytotoxicity was detected by CCK-8 combined with lactate dehydrogenase (LDH) Assay Kit, and the morphological changes of HEK293 cells were observed under an inverted phase contrast microscope. RESULTS: Species identification indicated that the four wild mushrooms were Butyriboletus roseoflavus, Boletus edulis, Russula virescens and Amanita manginiana. Cytotoxicity was found only in Amanita manginiana. The raw extracts showed cytotoxicity at the mass concentration of 0.1 mg/mL, while the boiled extracts and the boiled followed by enzymolysis extracts showed obvious cytotoxicity at the mass concentration of 0.4 mg/mL and 0.7 mg/mL, respectively. In addition to the obvious decrease in the number of HEK293 cells, the number of synapses increased and the refraction of HEK293 cells was poor after the intervention of Amanita manginiana extracts. CONCLUSIONS The extracts of Amanita manginiana involved in this YNSUD case has obvious cytotoxicity, and some of its toxicity can be reduced by boiled and enzymolysis, but cannot be completely detoxicated. Therefore, the consumption of Amanita manginiana is potentially dangerous, and it may be one of the causes of the YNSUD.
Humans ; HEK293 Cells ; Sincalide ; China ; Amanita ; Death, Sudden