ObjectiveTo investigate the mechanism of ferroptosis mediated by long-chain acyl-CoA synthetase 4 （ACSL4） signalling pathway in rats with coronary heart disease with blood stasis syndrome and the intervention effect of Xuefu Zhuyutang. MethodsSPF male SD rats were randomly divided into normal group， sham-operation group， model group， trimetazidine group （5.4 mg·kg^-1）， low-， medium-， and high-dose group （3.51， 7.02，14.04 g·kg^-1） of Xuefu Zhuyutang. The coronary artery left anterior descending ligation method was used to prepare a model of coronary heart disease with blood stasis syndrome， and continuous treatment for 7 d was conducted， while the sham-operation group was only threaded and not ligated. The general macroscopic symptoms of the rats were observed， and indicators such as electrocardiogram， echocardiography， and blood rheology were detected. The pathological morphology of myocardial tissue was observed by hematoxylin-eosin （HE） staining， and the changes in mitochondria in myocardial tissue were observed by transmission electron microscopy. The level of iron deposition in myocardial tissue was observed by Prussian blue staining. The levels of 12-hydroxyeicosatetraenoic acid （12-HETE） and 15-HETE were detected in serum by enzyme-linked immunosorbent assay. A biochemical colourimetric assay was used to detect the levels of Fe²⁺， lipid peroxidation （LPO）， glutathione （GSH）， and T-GSH/glutathione disulfide （GSSG） in myocardial tissue. DCFH-DA fluorescence quantitative assay was employed to detect the levels of reactive oxygen species （ROS）. Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was adopted to detect the protein and mRNA expressions of glutathione peroxidase 4 （GPX4）， ferritin heavy chain 1 （FTH1）， ACSL4， and ly-sophosphatidylcholine acyltransferase3 （LPCAT3） in myocardial tissue. ResultsCompared with those in the normal group， the rats in the model group were poor in general macroscopic symptoms. The electrocardiogram showed widened QRS wave amplitude and increased voltage， bow-back elevation of the ST segments， elevated T waves， J-point elevation， and accelerated heart rate. Echocardiography showed a significant reduction in left ventricular ejection fraction （LVEF） and left ventricular fraction shortening （LVFS） （P<0.01）. Blood rheology showed that the viscosity of the whole blood （low， medium， and high rate of shear） was significantly increased （P<0.01）. HE staining showed an abnormal structure of myocardial tissue. There was a large area of myocardial necrosis and inflammatory cell infiltration and a large number of connective tissue between myocardial fibers. Transmission electron microscopy showed that the mitochondria were severely atrophy or swelling. The cristae were reduced or even broken， and the matrix was flocculent or even vacuolated. Prussian blue staining showed that there were a large number of iron-containing particles， and the iron deposition was obvious. The content of 12-HETE and 15-HETE in the serum was significantly increased （P<0.01）. The content of Fe²⁺， LPO， and ROS in myocardial tissue was significantly increased （P<0.01）. The content of GSH was significantly decreased （P<0.01）， and T-GSH/GSSG was decreased （P<0.01）. The protein and mRNA expressions of GPX4 and FTH1 in myocardial tissue were both significantly decreased （P<0.05， P<0.01）， while those of ACSL4 and LPCAT3 increased significantly （P<0.01）. Compared with the model group， the general macroscopic symptoms and electrocardiogram results of rats in low-， medium- and high-dose groups of Xuefu Zhuyutang were alleviated， and the differences in LVEF/LVFS ratios were all significantly increased （P<0.05， P<0.01）. The differences in whole-blood viscosity （low， medium， and high rate of shear） were all significantly decreased （P<0.01）. The results of HE staining and transmission electron microscopy showed that the morphology， structure， and mitochondria of cardiomyocytes were improved. The content of 12-HETE and 15-HETE in serum was reduced to different degrees in low-， medium-， and high-dose groups of Xuefu Zhuyutang （P<0.05， P<0.01）. The content of Fe²⁺， LPO， and ROS was significantly reduced in the medium- and high-dose groups of Xuefu Zhuyutang （P<0.05， P<0.01）， and the content of GSH and T-GSH/GSSG was significantly increased （P<0.05， P<0.01）. The protein and mRNA expressions of GPX4 and FTH1 were significantly increased to varying degrees in the medium- and high-dose groups of Xuefu Zhuyutang （P<0.05， P<0.01）， and ACSL4 and LPCAT3 were decreased to different degrees in the low-， medium-， and high-dose groups of Xuefu Zhuyutang （P<0.05， P<0.01）. ConclusionXuefu Zhuyutang can regulate iron metabolism and anti-lipid oxidation reaction to mediate ferroptosis through the ACSL4 signalling pathway， thus exerting a protective effect on rats with coronary heart disease with blood stasis syndrome.

ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

Objective:To analyze the current situation of myopia and its related factors among primary school students in a certain district of Beijing City in 2022, and provide a basis for the risk assessment of myopia among primary school students.Method:In June 2022, a cluster sampling method was used to include 376 third-grade students from a primary school in a certain district of Beijing. A questionnaire survey was conducted to collect basic information about students, including eye usage habits, reading and writing postures, and parents′ myopia conditions. The examination of students′ distant visual acuity and refractive status was performed. A multivariate logistic regression model was used to analyze the related factors of myopia occurrence.Results:The age of 376 primary school students was (8.87±0.417) years old, with 48.40% (182) being male. A total of 196 myopia cases were identified, with a myopia rate of 52.13%. The results of the multivariate logistic regression model analysis showed that students who sometimes read while lying down ( OR=2.003, 95% CI: 1.128-3.555), often read while lying down ( OR=18.853, 95% CI: 4.512-78.778), had outdoor activity time less than 120 minutes per day ( OR=4.937, 95% CI: 2.4464-9.892), were engaged in indoor break activities ( OR=4.995, 95% CI: 2.773-8.996), performed eye exercises less than once per day ( OR=8.710, 95% CI: 4.464-16.995), had a reading distance from the book less than 30 cm ( OR=5.098, 95% CI: 2.410-10.787), occasionally maintained a fist distance from the edge of the desk ( OR=1.918, 95% CI: 1.086-3.385), and had high school desks and tables ( OR=5.325, 95% CI: 1.465-19.359) could have a higher risk of myopia occurrence, compared with those who never read while lying down, had outdoor activity time more than 120 minutes per day, maintained outdoor break activities, performed eye exercises more than once per day, had a reading distance from the book more than 30 cm, always maintained a fist distance from the edge of the desk, and had short school desks and tables. Conclusion:The incidence rate of myopia among primary school students in a certain district of Beijing City. in 2022 is relatively high. The occurrence of myopia is related to insufficient outdoor activity time and poor eye usage habits.

Objective:To analyze the current situation of myopia and its related factors among primary school students in a certain district of Beijing City in 2022, and provide a basis for the risk assessment of myopia among primary school students.Method:In June 2022, a cluster sampling method was used to include 376 third-grade students from a primary school in a certain district of Beijing. A questionnaire survey was conducted to collect basic information about students, including eye usage habits, reading and writing postures, and parents′ myopia conditions. The examination of students′ distant visual acuity and refractive status was performed. A multivariate logistic regression model was used to analyze the related factors of myopia occurrence.Results:The age of 376 primary school students was (8.87±0.417) years old, with 48.40% (182) being male. A total of 196 myopia cases were identified, with a myopia rate of 52.13%. The results of the multivariate logistic regression model analysis showed that students who sometimes read while lying down ( OR=2.003, 95% CI: 1.128-3.555), often read while lying down ( OR=18.853, 95% CI: 4.512-78.778), had outdoor activity time less than 120 minutes per day ( OR=4.937, 95% CI: 2.4464-9.892), were engaged in indoor break activities ( OR=4.995, 95% CI: 2.773-8.996), performed eye exercises less than once per day ( OR=8.710, 95% CI: 4.464-16.995), had a reading distance from the book less than 30 cm ( OR=5.098, 95% CI: 2.410-10.787), occasionally maintained a fist distance from the edge of the desk ( OR=1.918, 95% CI: 1.086-3.385), and had high school desks and tables ( OR=5.325, 95% CI: 1.465-19.359) could have a higher risk of myopia occurrence, compared with those who never read while lying down, had outdoor activity time more than 120 minutes per day, maintained outdoor break activities, performed eye exercises more than once per day, had a reading distance from the book more than 30 cm, always maintained a fist distance from the edge of the desk, and had short school desks and tables. Conclusion:The incidence rate of myopia among primary school students in a certain district of Beijing City. in 2022 is relatively high. The occurrence of myopia is related to insufficient outdoor activity time and poor eye usage habits.

Objective: To evaluate health risks of five heavy metals in drinking water of rural schools in Henan Province, so as to provide scientific basis for drinking water safety in rural schools. Methods: Totally 1 269 drinking water samples were collected and analyzed for five heavy metals (As, Cd, Cr 6+ , Pb, Hg) concentration in 106 cities and counties of Henan Province, and its spatial distribution characteristics were explored by geographic information system (GIS). Risk assessment method recommended by the United States Environmental Protection Agency (U.S. EPA) was applied to evaluate health risks from five heavy metals through oral ingestion. Results: The qualified rate of As concentration was 100%, and the qualified rates of Cd, Cr 6+ , Pb and Hg were 99.9% , 99.9%, 99.5%, 99.6%. The highest carcinogenic risk was As (3.05×10 -5 ), followed by Cr 6+ (2.73×10 -5 ), and the highest non carcinogenic risk was As (0.158 3), followed by Pb(0.041 7). The carcinogenic risk and non carcinogenic risk were all higher in girls than boys. The carcinogenic and non carcinogenic risks were differences in different regions( P <0.05). Conclusion Health risks of five heavy metals in drinking water for rural schools in Henan Province were within the acceptable risk level recommended by EPA. As, Cr 6+ and Pb were the main health risk factors in drinking water, and that might impact negatively the healthy growth of primary and middle school students. These should be future efforts of drinking water safety management.

Humans ; Pregnancy ; Female ; Umbilical Cord/diagnostic imaging* ; Pregnancy Outcome ; Ultrasonography ; Ultrasonography, Prenatal

Objective:To evaluate the effects of a booster immunization with a candidate tetanus toxoid, reduced diphtheria toxoid and acellular pertussis combined vaccine (Tdap) in a rat model after primary vaccination with diphtheria, tetanus, acellular pertussis and Sabin strain inactivated poliovirus combined vaccine (DTacP-sIPV) or diphtheria, tetanus, acellular pertussis, inactivated poliovirus and haemophilus type b combined vaccine (DTacP-IPV/Hib) for further preclinical study.Methods:Wistar rats were randomly divided into three groups and respectively immunized with a self-developed DTacP-sIPV, a marketed DTacP-IPV/Hib and normal saline at 0, 1, and 2 months of age. Serum levels of antibody against each component in each group were detected before immunization and after each dose. A booster dose of the candidate Tdap was given 10 months after primary immunization. Serum levels of antibody against each component in each group were detected before, 1 month and 6 months after the booster immunization.Results:One month after three doses of primary immunization, the geometric mean titers (GMT, Log2) of antibodies against diphtheria toxoid (DT), tetanus toxoid (TT), pertussis toxin (PT), filamentous hemagglutinin (FHA) and pertactin (PRN) in the DTacP-sIPV group were 17.41, 18.34, 18.11, 19.93 and 13.91, respectively, and the seroconversion rates of these components all reached 100%. Ten months after primary immunization, the GMTs of antibodies against DT, TT, PT, FHA and PRN decreased to 15.17, 14.26, 13.60, 14.51 and 10.39, respectively, and the seroconversion rates remained above 89%. One month after booster immunization, the GMTs of antibodies against DT, TT, PT and FHA in the DTacP-sIPV and DTacP-IPV/Hib groups were 16.49/17.26, 16.80/17.63, 16.70/17.74 and 18.48/19.26, respectively, and the seroconversion rates of these components all reached 100% with no significant difference between the two groups ( P>0.05). The GMTs of anti-PRN antibody in the DTacP-sIPV and DTacP-IPV/Hib groups were 13.07 and 11.00, and the seroconversion rates were 100% and 88%, which were higher in the DTacP-sIPV group than in the DTacP-IPV/Hib group ( P<0.05). Six months after booster immunization, the GMTs of antibodies against DT, TT, PT, FHA and PRN in the DTacP-sIPV and DTacP-IPV/Hib groups decreased to 15.74/14.87, 15.07/15.14, 14.84/15.73, 16.62/16.37 and 11.44/9.96, respectively, and the seroconversion rates remained above 88%. Conclusions:Booster vaccination with the candidate Tdap vaccine induces humoral immune response following primary immunization with DTacP-sIPV or DTacP-IPV/Hib in the Wistar rat model, while the antibody titer decreases with time.

Objective:To investigate the correlation between lipoprotein(a)[Lp(a)]levels and parameters for left ventricular structure in elderly hypertensive patients.Methods:A total of 65 patients aged ≥65 years diagnosed with essential hypertension were enrolled in this retrospective analysis.Left atrial diameter dilation(LAD)was measured to evaluate the degree of cardiac involvement.According to the existence of LAD, patients were divided into the negative group(n=43)and the positive group(n=22). The differences in levels of blood lipids including Lp(a)and LAD as well as other echocardiographic indexes between the two groups were compared, and their correlations were analyzed.Results:Compared with the negative group, the duration of hypertension, LAD, and Lp(a)level increased in the positive group( P<0.05). LAD was correlated with age, duration of hypertension, left ventricular end diastolic diameter and Lp(a)level.The Lp(a)level was correlated with age and LAD( P<0.05). Logistic regression analysis showed that duration of hypertension( OR=1.060, 95% CI: 1.008-1.116)and Lp(a)level( OR=6.394, 95% CI: 1.038-39.396)were independent risk factors for LAD.The area under the receiver operating characteristic(ROC)curve of Lp(a)for LAD was 0.665(0.524, 0.806). Conclusions:Laboratory testing for serum lipoprotein(a) levels will help assess the degree of cardiac involvement in patients with hypertension and has practical value for the risk assessment of cardiac remodeling in elderly hypertensive patients.

Objective:To investigate differences in the value of three chronic kidney disease epidemiology collaboration(CKD-EPI)estimation formulas in the evaluation of renal function damage in elderly patients with hypertension.Methods:A total of 58 patients with essential hypertension over 65 years of age were enrolled from November 2018 to March 2019 in this retrospective analysis.Patients with abnormal results in one or more items out of serum creatinine(Scr), blood urea nitrogen and urine protein were included in the hypertensive renal damage group(the positive group, n=25)and patients without abnormal results were included in the simple hypertensive group(the negative group, n=33). We measured Scr and cystatin c(Cys C)levels in fasting blood for each group.The estimated glomerular filtration rates(eGFR Scr, eGFR Cys C and eGFR Scr-Cys C)were calculated using the CKD-EPI formulas. Results:There was no statistically significant difference in gender, age, body mass index, grade of hypertension, onset time of hypertension, total cholesterol and triglycerides between the two groups( P> 0.05). There were significant differences in Cys C, eGFR Scr, eGFR Cys C and eGFR Scr-Cys C between the two groups( P<0.05). Spearman correlation analysis showed that Cys C was positively correlated with Scr( r=0.427, P<0.01), serum urate( r=0.266, P<0.05)and blood urea nitrogen( r=0.357, P<0.01)and was negatively correlated with eGFR Scr( r=-0.369, P<0.01), eGFR Cys C( r=-0.982, P<0.01)and eGFR Scr-Cys C( r=-0.845, P<0.01). Of the three formulas, eGFRScr-Cys C had the largest area under the receiver operating characteristic(ROC)curve(AUC)and its value was 0.707(0.565, 0.849). Conclusions:For the three estimation formulas, CKD-EPI Scr-Cys C has slightly better diagnostic efficiency than the others and may help improve the diagnosis of renal damage in elderly patients with hypertension.