The effects of pH, dosage, and the initial mass concentration of each treatment solution upon adsorption of F‐ were determined. Kinetic and thermodynamic models were applied to reveal the mechanism of defluoridation, and an orthogonal experiment was designed to obtain the optimal combination of conditions. The results show that the surfaces of CB, BC, and RHB have an irregular pore structure and rough surface, whereas DA has a rich pore structure, clear pores, large specific surface area, and high silica content. With regard to the adsorption process for F‐, DA has an adsorption complex electron interaction; that of CB, BC, and RHB occur mainly via ion exchange with positive and negative charges; and CB on F‐ relies on chemical electron bonding adsorption. The maximum adsorption capacity of DA can reach 32.20 mg/g. When the mass concentration of fluoride is 100 mg/L, the pH value is 6.0 and the dosage is 4.0 g/L; the adsorption rate of F‐ by DA can reach 91.8%. Therefore, we conclude that DA soil could be used as an efficient, inexpensive, and environmentally friendly adsorbent for fluoride removal, perhaps providing an empirical basis for improving the treatment of fluorine-containing water in the future.Water containing high concentrations of fluoride is widely distributed and seriously harmful, largely because long-term exposure to fluoride exceeding the recommended level will lead to fluorosis of teeth and bones. Therefore, it is imperative to develop cost-effective and environmentally friendly adsorbents to remove fluoride from polluted water sources. In this study, diatomite (DA), calcium bentonite (CB), bamboo charcoal (BC), and rice husk biochar (RHB) were tested as adsorbents to adsorb fluoride (F Current2-nH2O), mainly derived from aquatic diatom plant skeletons. Due to its high porosity, large surface area, high permeability, low density, small particle size, heat resistance, and chemical stability  in water and to screen for material’s best enabling adsorption for fluoride. A second objective was to investigate the adsorption mechanism for fluoride in water in high-fluorine areas to provide a theoretical basis for the treatment of fluoride-containing water.In summary, many studies have examined the use of adsorption materials to remove fluoride in water, but the adsorption capacity and action mechanism of different adsorption materials are likely to differ. Accordingly, in this study, diatomite (DA), calcium bentonite (CB), bamboo biochar (BC), and rice husk biochar (RHB) were selected to systematically study their adsorption properties for fluoride . Their crystal structures were characterized by X-ray diffraction , and their surface structural groups were analyzed by Fourier transform infrared spectroscopy .‐). The experimental water was deionized; the buffer solutions used for the experiment were sodium citrate dihydrate and sodium nitrate . The instruments used in the experiment were a JA1003 electronic balance, a PHS-3C type pH meter, a BS-MS thermostatic oscillator, an L550 Xiang Yi centrifuge, a CM-230 laboratory pure water treatment system, a PXS-270 fluorometer, and a JB-10 magnetic stirrer.Sodium fluoride (GR) was used to simulate aqueous fluoride , and the adsorption experiment was carried out at 25°C for 120 min in a constant temperature shaker at 200 rpm. After centrifugation at 4,000 r/min for 10 min, 10 ml of the supernatant was centrifuged through a 0.45 μm filter membrane, and the remaining F‐ mass concentration in the solution was determined using a PXS-270 fluoride ion-selective electrode. Each adsorbent was tested for each F‐ concentration in triplicate, and relative standard deviations of duplicate samples were less than 5.0%.The initial mass concentration gradient of F‐ (100 mg/L) sorption by each adsorbent (0.10 g) were determined with 25 ml solutions for 5, 10, 20, 40, 60, 120, 240, 360, 480, and 720 min at 25 °C in the solution at pH 6. After centrifugation at 4,000 r/min for 10 min, 10 ml of the supernatant was centrifuged through a 0.45 μm filter membrane, and the remaining F‐ mass concentration in the solution was measured.The kinetic parameters of F‐ solution (100 mg/L) was added into 50 ml centrifuge tubes, and its pH was adjusted to 3.0, 4.0, 5.0, 6.0, 7.0, or 8.0. To each tube, 0.10 g of DA was added, and all tubes were stored at 25°C. The adsorption experiment was the same as given in Section 2.3.1. After centrifugation, 10 ml of the supernatant was taken and passed through a 0.45 μm filter membrane to determine the remaining F‐mass concentration in the solution., About 25 ml of an F‐ (100 mg/L) in solution was added to each tube. The adsorption experiment was the same as given in Section 2.3.1. After centrifugation at 4,000 r/min for 10 min, 10 ml of the supernatant was centrifuged through a 0.45 μm filter membrane and then the remaining F‐ mass concentration in the solution was measured.The DA was weighed at 0.04, 0.08, 0.10, 0.15, 0.20, and 0.30 g in 50 ml centrifuge tubes, and 25 ml of a mass concentration of F‐ solution (B), and adsorbent dosage (C) on the adsorption of F‐ in water. Three levels of each factor were selected for the three-factor, three-level orthogonal test of L934.According to the design method of the orthogonal test, the factors selected for investigation were as follows: adsorption pH (A), the mass concentration of the FThe removal rate is calculated as:‐ in solution at the adsorption equilibrium (mg/L), andwhereThe adsorption volume is calculated as follows:‐ in the solution at the adsorption equilibrium (mg/L), ‐ at the adsorption equilibrium (mg/g).where The two adsorption fitting models are as follows. The Langmuir model equation is The Freundlich model equation is ‐ in the solution at the adsorption equilibrium (mg/L), ‐ at the adsorption equilibrium (mg/g), n > 1, linear adsorption for n = 1, and non-preferential adsorption for n < 1.where The quasi-first-order kinetic equation isThe quasi-second-order kinetic equation ist (mg/g), where 2. The sharp diffraction peaks at around 26° can be attributed to quartz impurities in the materials, while the wider diffraction peaks at around 35° correspond to amorphous Al2O3  and the surface hydroxyl group. All four materials have vibrational peaks around 1,634 cm−1, probably from the bound water or the surface hydroxyl group. The peaks of DA, CB, BC, and RHB located at 1,100 cm−1 and 538 cm−1 are of siloxane groups (Si-O-Si-), and the peak at 792 cm−1 is attributable to the Al-O absorption band  was higher than that of BC and RHB. It can be concluded that the magnitude of adsorption performance of the four materials is DA > CB > BC > RHB. The adsorption rate of Fon of F‐ . The higon of F‐ . When thion rate . For the‐ processes of the four materials, and their fitted parameters are in R2 values of up to 0.9930, indicating that the adsorption process of DA for F‐ is an adsorption–complex electron interaction, surface adsorption, and multi-molecular layer adsorption. When applied to CB and BC, both models fit better, indicating that the adsorption process of F‐ by CB and BC is unilamellar and multilamellar co-adsorption, mainly based on ion exchange involving positive and negative charges. For RHB, the Langmuir model fits better than the Freundlich model, with an R2 value of 0.9128, indicating that the adsorption process of F‐ by RHB is unilamellar. Its adsorption mechanism is mainly based on ion exchange involving positive and negative charges. Similar results were reported by The Freundlich and Langmuir models were used here to describe the adsorption of F‐ by the four materials over time. Clearly, the adsorption amount increased with the elapsed time, and the maximum adsorption amount was reached at around 120 min. The materials’ adsorption amounts were ranked as follows: DA > CB > BC > RHB; the maximum adsorption amount of DA reached 22.300 mg/g, twice as much as that found for the other three materials. Beyond 120 min, the adsorption amount did not change and adsorption had reached equilibrium. ‐ rapidly occupies the active sites on the adsorbent’s surface, and its adsorption capacity increases  and biochar adsorbents. The characterization of four adsorbent materials was investigated by FEI-SEM, XRD, and FT-IR. The DA material has an excellent surface structure and is rich in oxygen-containing functional groups vis-à-vis the other three tested materials, conferring to it a strong adsorption force for F


Babesia microti (B. microti) is the dominant species responsible for human babesiosis, which is associated with severe hemolytic anemia and splenomegaly because it infects mammalian erythrocytes. The actual prevalence of B. microti is thought to have been substantially underestimated.B. microti-infected erythrocytes, and parasitemia was subsequently measured by calculating the proportion of infected erythrocytes. The ultrastructure of infected erythrocytes was observed using scanning and transmission electron microscopes. Quantifying phosphatidylserine (PS) exposure, oxidative stress, intracellular Ca2+, and erythropoiesis of erythrocytes were done using flow cytometry. The physiological indicators were analyzed using a Mindray BC-5000 Vet automatic hematology analyzer.In this study, Bagg’s albino/c (BALB/c) mice were intraperitoneally injected with B. microti-infected group. The percentage of annexin V-positive erythrocytes and the levels of reactive oxygen species (ROS) in the erythrocytes were higher in the B. microti-infected group than in the control group at 10 dpi. Significant splenomegaly and severe anemia were also observed following B. microti infection. The parasitemia level in the B. microti-infected splenectomized group was higher than that of the B. microti-infected sham group. The population of early erythroblasts increased, and the late erythroblasts decreased in both the bone marrow and spleen tissues of the B. microti-infected group at 10 dpi.Of note, 40.7 ± 5.9% of erythrocytes changed their structure and shrunk in the B. microti-infected group. This study revealed for the first time that B. microti could also induce eryptosis. At the higher parasitemia phase, the occurrence of severe anemia and significant changes in the abundance of erythroblasts in B. microti-infected mice group were established. The spleen plays a critical protective role in controlling B. microti infection and preventing anemia. B. microti infection could cause a massive loss of late erythroblasts and induce erythropoiesis.PS exposure and elevated ROS activities were hallmarks of eryptosis in the  Macroph control . However2.2.1.Babesia microti strain ATCC®PRA-99TM used in this study was obtained from the Institute of Laboratory Animal Sciences, Chinese Academy of Medical Sciences (CAMS). This strain was maintained through serial passage in i.p. infected Bagg’s albino/c (BALB/c) mice. Infection was confirmed by observing for the presence of parasites in thin blood smears using an optical microscope at 3 or 5 dpi.2.2.7B. microti-infected erythrocytes per milliliter. The mice in infected groups were intraperitoneally injected with 107 infected erythrocytes in a volume of 0.2 ml. To ensure all mice received equal numbers of viable parasites, they were infected with equal parasite inoculum at the same time. Blood for smears was collected daily from the tail snip for 30 days post-infection (dpi) to evaluate the parasitemia level. The proportion of infected erythrocytes was calculated as previously described , aged 6–8 weeks, were purchased from the Shanghai Laboratory Animal Center (China). The housing and maintenance of the rodents complied with national regulations. Orbital blood samples were collected from the infected mice and diluted with sterile saline to 5 × 10escribed , based o2.3.Orbital sinus blood samples were collected in ethylenediaminetetraacetic acid at 10 dpi. The blood was immediately fixed for 2 h by electron microscopy fixative at room temperature. The preservation and transportation temperature was 4°C. Each sample was washed three times for 15 min with 0.1 M phosphate buffer (PB) (pH 7.4) and stained for 1–2 h at room temperature using 1% osmium tetroxide in 0.1 M PB (pH 7.4). The cells were dehydrated for 15 min in serial ethanol concentrations of 50%, 70%, 80%, 90%, and 95% ethanol, twice in 100% ethanol, and finally in isoamyl acetate. The cells were dried using a critical point dryer  and attached to metallic stubs using carbon stickers and sputter-coated with gold for 30 s. The cells were observed using an SEM  and a TEM .2.4.Mouse blood cells were washed twice using Ringer’s solution supplemented with 5 mM calcium chloride. To detect FITC annexin V-positive cells, the erythrocytes were suspended in an annexin-binding buffer  with FITC annexin V  and incubated for 15 min at room temperature. Well mixing was performed by pipetting. Finally, erythrocytes were diluted five times in the annexin-binding buffer before analysis in the FACSVerse flow cytometer  at an excitation wavelength of 488 nm (blue laser) and emission wavelength of 530 nm.2.5.The oxidative stress level was determined using 2′, 7′-dichlorodihydrofluorescein diacetate . Briefly, the erythrocytes (4 μl) were mixed in 1 ml Ringer’s solution, from which 150 μl of the cell suspension was centrifuged at 1,600 rpm for 3 min at room temperature. The collected cells were stained with DCFDA (10 μM) in Ringer’s solution at 37°C for 30 min and washed three times with 150 μl of Ringer’s solution. The DCFDA-loaded erythrocytes were then resuspended in 200 μl Ringer’s solution . The reactive oxygen species (ROS)-dependent fluorescence intensity was measured at an excitation wavelength of 488 nm and an emission wavelength of 530 nm.2.6.Calcium ion influx was evaluated using Fluo-3 Am  staining. Erythrocytes were suspended in 200 μl Ringer’s solution supplemented with Fluo-3 Am (1 μM) and incubated at 37°C for 30 min. Thereafter, the RBCs were stained with Fluo-3 Am, rinsed, and resuspended in Ringer’s solution, and analyzed using a 488 nm blue laser and a 530 nm bandpass filter.2.7.Mice were anesthetized using ether and a small incision was made under the left costal margin before ligating the splenic vessels. For mice in the sham group, the spleens were exposed but not removed. Blood was collected from the orbital sinus into a microhematocrit tube every 5 days until 20 dpi. The blood was analyzed using a Vet automatic hematology analyzer  for animals. The physiological indicators measured included the RBC count, hemoglobin concentration (HGB), red cell distribution width standard deviation (RDW-SD), and red cell distribution width coefficient of variation (RDW-CV).2.8.Erythropoiesis was analyzed as previously described . The mou2.9.® software (version 2019) and GraphPad Prism (version 8.3.0). Continuous data were expressed as the mean ± SD. The difference between groups was analyzed using the student’s t-test, whereas analysis of variance (ANOVA) was used for multiple groups. Statistical significance was set at p < 0.05.Data were analyzed using Microsoft Excel3.3.1.B. microti-infected group had a significant increase in spurred erythrocytes that were irregularly distributed, variably sized, and with pointy projections off their surfaces -labeled Annexin V. Of note, the percentage of annexin V-positive erythrocytes was significantly higher in the ficantly .3.3.B. microti-infected group were 2.34 ± 0.09, 3.36 ± 0.13, 3.08 ± 0.08, and 2.74 ± 0.25 g while those of the control group were 1.77 ± 0.16, 1.72 ± 0.08, 1.70 ± 0.14, and 1.78 ± 0.08 g at 5, 10, 15, and 20 dpi  (B. microti infection (B. microti-infected sham and splenectomized groups (sham mice had the spleen while splenectomized mice had their spleens harvested) to assess the role of the spleen during B. microti infection. Parasitemia in both groups increased rapidly at 6 dpi and peaked at 10 or 11 dpi. Of note, 40.56 ± 4.14% of the erythrocytes from the B. microti-infected sham mice and 56.88 ± 3.97% of the erythrocytes from the B. microti-infected splenectomized mice were infected at 10 and 11 dpi, respectively. The peak parasitemia level of the B. microti-infected splenectomized group was higher than that of the B. microti-infected sham group and persisted for ~6 days. The difference in parasitemia level between the B. microti-infected sham and splenectomized groups at 11–20 dpi was statistically significant (p < 0.01). In the B. microti-infected sham and splenectomized groups, the parasites could not be detected in erythrocytes by 23 and 27 dpi, respectively. These results indicate that the spleen plays a defensive role in B. microti infection  . Notablynfection ,C. We hanfection .B. microti-infected sham and splenectomized mice compared to mice in the uninfected sham and splenectomized groups at 10 and 15 dpi. However, the RBCs and HGB of mice in the B. microti-infected splenectomized group were lower than those in the B. microti-infected sham group at 15 and 20 dpi  and a decrease in the late erythroblast population  in both the bone marrow and spleen tissues of the B. microti-infected group at 10 dpi  mice and nonobese diabetic SCID mice, in vivo models using BALB/c mice have shown that B. microti infection can resolve spontaneously after reaching peak parasitemia , National Institute of Parasitic Diseases of China CDC.PS and Y-CC: data curation and writing the original manuscript draft. PS, Y-CC, and M-XC: methodology. S-HC and J-XC: writing, review and project supervision. All authors contributed to the article and approved the submitted version.This project was funded by the Shanghai Municipal Health Commission (20194Y0046), the Shanghai Natural Science Foundation (21ZR1469900), the National Parasitic Resources Center, and the Ministry of Science and Technology (NPRC-2019-194-30).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The objective of this study was to evaluate the long-term surgical results of transposition of the great arteries with left ventricular outflow tract obstruction.We conducted a retrospective study of patients with transposition of the great arteries or double outlet right ventricle with left ventricular outflow tract obstruction undergoing biventricular repair between 1980 and 2017.One hundred and eleven patients were enrolled and classified into five groups: atrial switch (n = 20), arterial switch (n = 12), Nikaidoh (n = 7), Rastelli (n = 48), and REV operation groups (n = 24). Early mortality was highest in Nikaidoh group (29%). Median follow-up was 18.2 years. Long-term survival was by far lowest in Nikaidoh group and comparable among the other 4 groups. Freedom from reoperation at 20 years was lowest in Rastelli group (32.1%) due to right ventricular outflow tract-related reoperations. While having no recurrence of left ventricular outflow tract obstruction, the arterial switch operation group had a high proportion of substantial neo-aortic regurgitation (29%).The long-term survival was satisfactory regardless of the surgical technique except Nikaidoh group. The surgical option for transposition of the great arteries with left ventricular outflow tract obstruction should be selected based on the features of the respective procedures. The surgical treatment of transposition of the great arteries (TGA) with significant left ventricular outflow tract obstruction (LVOTO) continues to evolve. The Rastelli and atrial switch operations were first introduced as treatments for TGA with LVOTO in 1969 . The aorThis study aims to compare the long-term results of the five main procedures: the atrial switch, arterial switch, Nikaidoh, Rastelli, and REV operations, and presented the long-term features of each procedure.This study is a retrospective, cohort study with consecutive patients undergoing biventricular repair for TGA or TGA-type DORV with LVOTO at the Tokyo Women’s Medical University Hospital from 1980 to 2017. During this period, a total of 748 patients were diagnosed with TGA or a TGA type of DORV and underwent surgery. Of these patients, 111 patients with TGA/LVOTO were enrolled herein. This study was approved and monitored by the Tokyo Women’s Medical University’s research ethics committee  and the need for patient consent was waived due to the study’s retrospective nature. This study was performed in conformity with the Declaration of Helsinki. Medical records were reviewed, and the following data were retrieved and analyzed: basic demographic data, anatomical information, intraoperative data, and postoperative outcomes including catheter intervention, reoperation, and echocardiogram data.In this study, we reviewed the transition of the surgical strategies for TGA with LVOTO, identified the advantages and disadvantages of the main five procedures by evaluating the long-term surgical results of TGA with LVOTO, and presented a perspective view of the strategies for TGA with LVOTO.The type of definitive operation was selected for each patient based on the anatomic and hemodynamic characteristics during the study period Fig. . The ASOAll statistical analyses were performed with JMP Pro version 14 software . The normal or non-normal distribution of continuous variables was confirmed using the Shapiro–Wilk test. Data of continuous variables were presented as mean ± standard deviation for normal distribution and as median  for non-normal distribution. Categorical variables were presented as a proportion (frequency). Early mortality was defined as death before hospital discharge or within 30 days of the definitive operation. The survival time was estimated from the date of the definitive operation to the date of all-cause death or the last contact, and the freedom from reoperation time was estimated from the date of definitive operation to the date of reoperation. The endpoint of follow-up was defined as the date of death or the last contact. Actuarial survival rates were analyzed by the Kaplan–Meier curve. Valve diameter was expressed as Z-scores . LVOTO wA total of 111 patients were classified into five groups based on the type of definitive operations: the atrial switch operation (n = 20), ASO (n = 12), Nikaidoh (n = 7), Rastelli (n = 48), and REV operations (n = 24). The ASO group had the youngest mean operative age of 0.3 years and the Rastelli group had the oldest one of 5.3 years. Forty-six balloon atrial septostomy and 61 palliative operations were performed before the definitive operation. The category of LVOTO was valvular in 34 patients (31%), subvalvular in 30 (27%), atresia in 15 (14%), and subvalvular/valvular in 32 (29%). The most common type of LVOTO was small pulmonary diameter with valvular stenosis and left deviation of the infundibular septum with subvalvular stenosis. Preoperative median Z-scores of pulmonary diameter in valvular LVOTO for atrial switch operation, ASO, Nikaidoh operation, Rastelli operation, and REV operation groups were − 0.20 , 0.73 , − 0.64 , − 3.84 , and − 1.99 , respectively. Preoperative left ventricular shortening fraction was not notably different between the groups. Preoperative patient data were summarized in Table In atrial switch group, Senning n = 18) or Mustard n = 2) technique was used. In ASO group, the neo-PA was reconstructed by the original method (n = 2) or the Lecompte maneuver (n = 10), and the VSD was enlarged for intraventricular baffle in 5 TGA-type DORV patients. In Nikaidoh group, the RVOT was reconstructed using an autologous pericardial transannular patch. In Rastelli group, the RVOT was reconstructed with a xenograft roll conduit bearing handmade xenograft pericardial valves , an autologous pericardial roll conduit bearing handmade autologous pericardial valves , or an expanded polytetrafluoroethylene (e-PTFE) conduit with bulging sinuses and fan-shaped valves 2012–2017; n = 3) [; n = 3 [ or Musta techniquDuring the median follow-up of 18.2 7.3–25.0) years, 74 patients were alive, 28 died, and 9 were lost to follow-up. There were 8 early mortalities , which were all recorded in the 1980s or 1990s. Extracorporeal membrane oxygenation was introduced in four of these patients. There was no early mortality in REV group. The most frequent cause of these early mortalities was low output syndrome. The causes of death are shown in Table –25.0 yeaSurvival at 20 years after definitive operation was 78.5 ± 9.6% in atrial switch group, 75.0 ± 12.5% in ASO group, 42.9 ± 18.7% in Nikaidoh group, 75.5 ± 6.5% in Rastelli group, and 76.7 ± 10.8% in REV group  required postoperative catheter intervention: 2 in atrial switch, 5 in ASO, 1 in Nikaidoh, 14 in Rastelli, and 3 in REV. The most common catheter intervention for each group was pacemaker implantation in atrial switch operation group (2/2: 100%), percutaneous RVOT dilatation in Rastelli group (8/14: 57%), and percutaneous distal PA angioplasty in ASO (5/5: 100%), Nikaidoh (1/1: 100%), and REV (2/3: 67%) groups. Catheter ablation was performed for supraventricular tachycardia in 3 patients in Rastelli group.A total of 40 reoperations were conducted on 36 patients. Freedom from reoperation at 20 years after definitive operation was 87.7 ± 8.3% in atrial switch operation group, 61.1 ± 15.7% in ASO group, 100% in Nikaidoh group, 32.1 ± 8.4% in Rastelli group, and 85.2 ± 8.0% in REV group  and 4 in REV group. In Rastelli group, conduit stenosis was repaired by conduit replacement (n = 20) or direct right ventricle-to-PA anastomosis with patch angioplasty (n = 2), and infectious endocarditis was treated by bioprosthetic pulmonary valve replacement (n = 1). In REV group, RVOT was reconstructed with an autologous pericardial patch bearing a handmade autologous pericardial valve (n = 4). Overall, freedom from RVOT-related reoperation at 20 years after definitive operation was 33.1 ± 8.9% in Rastelli group, 90.2 ± 6.7% in REV group, and 100% in other groups , 1 residual VSD closure with concomitant tricuspid annuloplasty (ASO), and 3 tricuspid valve replacement for systemic atrioventricular valve regurgitation .Late echocardiography data were obtained from 73 of 83 survivors (88%). The median Z-score of valve diameter in the systemic ventricular outflow tract in atrial switch operation, ASO, Rastelli and REV groups was 2.0 (1.4–2.6), 1.7 (1.3–1.9), 0.2 (0.2–0.4), 2.5 (1.4–3.6), and 2.5 (1.4–3.6) respectively and the median flow velocity in the systemic ventricular outflow tract was 1.3 (1.1–1.4), 2.0 (1.6–2.1), 1.0 (0.9–1.1), 1.3 (0.9–2.0), and 1.3 (0.9–1.7) m/s respectively. A high incidence of substantial neo-aortic regurgitation was found in ASO group (2/7: 29%). A proportion of the substantial pulmonary regurgitation was comparable among Nikaidoh, Rastelli, and REV groups (67% vs. 58% vs. 63%). The median systemic ventricular shortening fraction in atrial switch operation, ASO, Nikaidoh operation, Rastelli operation, and REV operation groups was 0.20 (0.17–0.22), 0.37 (0.31–0.38), 0.31 (0.26–0.33), 0.28 (0.24–0.34), and 0.27 (0.26–0.29), respectively. No patients had Class III or IV on the New York Heart Association functional classification.Since the initial atrial switch operation  and RastThe surgical selection for TGA with LVOTO greatly changed after the introduction of the ASO, however, its use remains controversial. A relatively wide pulmonary annulus with no or small VSD made the ASO and atrial switch operation possible in our study. A reasonable threshold of the ASO  provided a favorable outcome with the mean late neo-aortic Z-score of 1.7 without LVOTO recurrence. However, our threshold of the indication for ASO might be extended to a pulmonary diameter Z-score of between − 1.5 and − 1.0 as a preoperative pulmonary valve Z-score of less than − 1.7 has been reported to be an independent predictor of recurrent LVOTO after the ASO for TGA/LVOTO , 12, andThe Nikaidoh operation is a technically demanding operation with the risks of destabilizing the aortic valve or kinking/compressing the proximal coronary arteries. Due to frequent coronary troubles, the Nikaidoh operation was abandoned in 1996 at our institution. Nikaidoh group in our study experienced the highest early mortality and consequently the lowest long-term survival of 42.9% among the groups while presenting with a low rate of late mortality and reoperation. The Nikaidoh operation has been reported to provide low early mortality of 3–5% as well as superior long-term outcomes in some reports . AdditioRVOT obstruction or pulmonary regurgitation is the most serious complication after the definitive operation requiring RVOT reconstruction. Similar to previous reports , 16, 18,The presence of LVOTO in TGA patients was revealed to be an independent risk factor after the ASO , 20, 21.This study has certain limitations: this was a retrospective, single-center study and biases were present owing to a lack of homogeneity, which can make group comparisons inaccurate; and finally, there was a lack of cases with follow-up. As this was a long and wide retrospective study, our study also included bias regarding postoperative management, surgical techniques, and uncommon surgical strategies.The 40-year surgical results at our institution revealed the unique advantages and disadvantages of the respective procedures. There were no significant differences in survival between the groups except for Nikaido group. Although having a low incidence of LVOTO recurrence and the best systemic ventricular function, ASO group presented a high proportion of late substantial neo-aortic regurgitation. In terms of avoidance of reoperation, the REV operation could be one of the optimal procedures for patients with severe LVOTO unsuitable for ASO and aortic root translocation.


Ellochelon vaigiensis (squaretail mullet) adapts to a wide salt spectrum, grows quickly and is easy to raise with other species, so it is the object of attention in aquaculture. Information on the biology and ecology of the species, diet, in particular, is still scattered. Here, we explore the feeding habit, feeding intensity, and food composition of the squaretail mullet. A total of 942 fish  were collected from November 2020 to October 2021 at four coastal sites in the Mekong Delta, Vietnam. The squaretail mullet is an algi‐omnivorous fish, as their relative gut length (1.81) falls into the range 1–3, and the diet is mostly based on algae. The feeding intensity is high due to the high value of the fullness index (4.39 ± 0.08%). The fullness index did not vary by sampling site and month, while RGL and FI changed depending on sex. Bacillariophyta (49.13%), detritus derived from organic matter (30.37%), and Cyanophyta (18.39%) are the dominant food items in the diet composition of E. vaigiensis, in which detritus is the most important food with the highest IRI index. Besides, Euglenophyta (1.00%), Chlorophyta (0.95%), Paramecium (0.06%), Copepoda (0.04%), Rotatoria (0.03%), Polychaeta (0.02%), and Cladocera (0.01%) are also recorded and ranked based on their biovolume. Some differences in diet composition are observed between immature and mature at different seasons and their interactions. Our results increase the knowledge about the feeding ecology of squaretail mullet and can help the sustainable management of this commercially important fish species. The results denote that squaretail mullet is an algi‐omnivorous fish, as their relative gut length (1.81) falls into the range 1‐3, and the diet is mostly based on algae. The feeding intensity of this mullet is high due to the high value of the fullness index (4.39 ± 0.08%). The results obtained from this study increase the knowledge about the feeding ecology of squaretail mullet and can help the sustainable management of this commercially important fish species in the MD. Moreove Hyslop, . The metTo determine the importance of types of food in the diet composition, we calculated the percentage of the important relative index (%IRI) of each food item as follows: %IRI = %O × (%N + %V)  Windell, .2.2The normal distribution of RGL and FI was verified by Kolmogorov–Smirnov test Kim, . Since tLm) was used to divide fish into the immature group (TL < Lm) and the mature group (TL ≥ Lm). The male and female Lm was calculated as P = 1/(1 + exp[−r × (TL‐Lm)])   Zar, .Dietary changes by sex, body size, season, and site based on biovolume percentage of foods were analyzed in the software PRIMER v.6.1.11 with PERMANOVA+v.1.0.1, according to Dinh et al. . After i33.1KS = 0.04, p = .004). The overall relative gut length (RGL) was 1.81 ± 0.01, ranging from 1 to 3; therefore, the species was an omnivore . The RGL exhibited significant differences according to sex but not by body size, season, and sites. Specifically, female RGL (1.89 ± 0.03) was significantly higher than that of males (1.73 ± 0.03) . Immature and mature fishes exhibited RGL values not statistically different . This can suggest that the feeding habit did not change from juvenile to adult. The feeding pattern of the squaretail mullet in the dry season (1.81 ± 0.03) was not significantly different from that of the wet season (1.80 ± 0.02) . In addition, squaretail mullets did not change their feeding habits at the four sampling sites as their RGL values ranged from 1 to 3  . The RGL was independent on the interactions of sex × body size , sex × season , sex × site , body size × season , body size × site , season × site .The 3.2KS = 0.106, p < .01). This value of the squaretail mullet was relatively high, about 4.39 ± 0.08%, indicating that the mullet has a high feeding intensity. The differences in body size , season , and sites  did not affect FI values. The FI of females (4.20 ± 0.12%) was statistically lower than that of males (4.55 ± 0.12%) , whereas the FI of immature (4.53 ± 0.22%) was equivalent to mature (4.04 ± 0.14%). Similarly, this value did not differ between the dry season (4.26 ± 0.21%) and the wet season (4.29 ± 0.14%). The analysis results also showed that FI fluctuated not significantly amongst Ben Tre (4.71 ± 0.39%), Tra Vinh (4.27 ± 0.17%), Soc Trang (3.84 ± 0.20%), and Bac Lieu (4.32 ± 0.25%). The interactions of site × season  and sex × body size  effected on FI. The remaining interactions showed no significant effect on FI such as site × sex , site × body size , sex × season , and season × body size .The distribution of FI values was not normal   and sampling season  significantly affected diet composition. The result of MDS visualized the food composition according to body size and seasons and revealed that Bacillariophyta, detritus derived from organic matter, and Cyanophyta were the three main foods , whereas they ate other foods equally. In dry season, this mullet consumed more detritus , Cyanophyta , Chlorophyta , and Polychaeta  than in wet season. Otherwise, the squaretail mullet ate more Bacillariophyta  and Euglenophyta . No differences were found in the other prey items between dry and wet seasons.The result of the Mann–Whitney test revealed that immature fish ingested more Copepoda (0.12 ± 0.03%) than mature fish (0.05 ± 0.01%) ; sex × season ; and season × site . The results of the MDS graphs visualized the influence of the above interactions on the diet composition of mullet; and Bacillariophyta, detritus derived from organic matter, and Cyanophyta were still the three main food , whereas, in the mature group, the male and female consumed the same food except for Bacichloropyta . Considering the interaction between sex and season, detritus , Bacillariophyta , Cyanophyta , Chlorophyta , Euglenophyta , and Polychaeta  were the main cause of variation in diet of male in dry and wet season, whereas, in female, Detritus , Bacillariophyta , Cyanophyta , and Euglenophyta  caused a seasonal variation in their diet composition. In the wet season, the dietaries of this mullet at four sampling sites were not statistically different . However, in the dry season, their food composition differs according to the sampling sites, due to the variability of the following food items: detritus , Bacillariophyta , Cyanophyta , Chlorophyta , and Euglenophyta . Thus, it could be concluded that the diet of this fish changed by sex at different fish sizes in the dry and wet seasons at each sampling site. Conversely, the diet composition was not dependent on the interaction of sex × site , body size × season , and body size × site .Refer to the interaction between sex and body size on the food spectrum of mullet, in the immature group, Chlorophyta was the only food that differed between males and females  indicated the feeding intensity of squaretail mullet was strong. A similar result was found in E. vaigiensis consumed diatom, algae, detritus, polychaetes, mollusks, and crustaceans together with sand and mud. Liza vaigiensis, in the study of Hajisamae et al. , algae (2.31 and 9.85%), desmids, plant materials, zooplankton (1.92 and 18.23%), detritus, sand grains, and fishes . Notably, in the study of Wijeyaratne and Costa , the main food item of L. vaigiensis changed by month, but serpulid polychaetes were always the essential food item. Specifically, detritus was the main food item from April to December, while serpulid polychaetes were dominant from January to March in squaretail mullet with a total length of 5–10 cm; whereas, in July and October detritus was the major item and in August, sand particles were the most in size group of 20–25 cm.Sex differences in food selection may be due to random selection or to different energy requirements between males and females, but within the scope of this study, the primary and preferred foods item for both sexes were not different. It was evident that males and females of squaretail mullet tended to feed more actively on Bacillariophyta, detritus, and Cyanophyta than other food items. Similarly, d Costa , the ma5E. vaigiensis in this study provided systematic information on the nutritional biology that is useful in the future artificial culture of this species.The squaretail mullet was algi‐omnivorous, and most of its food items were algae‐based, consisting of Bacillariophyta, detritus derived from organic matter, Cyanophyta, Euglenophyta, Chlorophyta, Paramecium, Copepoda, Rotatoria, Polychaeta, and Cladocera. The diet composition changed concerning sex and season. The feeding intensity of this mullet was relatively high and tended to decrease in the breeding season and increase in the post‐breeding season. The knowledge about the food and feeding habits of Quang Minh Dinh: Conceptualization ; data curation ; funding acquisition ; investigation ; methodology ; project administration ; resources ; validation ; writing – original draft ; writing – review and editing . Ngon Trong Truong: Investigation ; methodology ; writing – original draft ; writing – review and editing . Ton Huu Duc Nguyen: Conceptualization ; data curation ; investigation ; resources ; writing – original draft ; writing – review and editing . Tran Thi Huyen Lam: Conceptualization ; investigation ; methodology ; resources ; validation ; writing – original draft ; writing – review and editing . Tien Thi Kieu Nguyen: Investigation ; methodology ; validation ; writing – original draft ; writing – review and editing . Dung Quang Le: Investigation ; validation ; writing – original draft ; writing – review and editing . Simon Kumar Das: Investigation ; validation ; writing – original draft ; writing – review and editing .This work was funded by VINGROUP and supported by Vingroup Innovation Foundation (VINIF) under project code VINIF.2020.DA01.The authors declare that they have no competing interests.


Competitive sports involve physiological, technical and psychological skills, which influence directly on individuals’ performance. This study aims to investigate the levels of perceived stress and Heart Rate Variability (HRV) before and after matches with victory and defeat in professional eSports athletes. Our hypothesis was that the winners would have better autonomic and stress responses after match, thus corroborating the literature on neurocardiac connections. Fifty male eSport players were selected players from 10 different Brazilian teams. The experiment was carried out in 2 sessions. Firstly, after signing the informed consent form, 24 h before the game, anthropometric, physical activity levels and time of expertise data were recorded only for sample characterization and the players were familiarized with the perceived stress scale—10 (PSS-10) and the HRV measurements. Secondly, players performed the PSS-10 and HRV recording at rest by 10 min 60 and 30 min before the game  and 10 min after the end of the game. Overall, concerning PSS-10 our findings show that VG had significant reduced scores in post-game time compared to baseline (BL) and pre-game times, while DG had significant increased scores in post-game time compared to BL and pre-game times. Regarding HRV, our results demonstrate that VG had significant increase in RR, SDNN, rMSSD, pNN50 and HF, and significant decrease in LF and LF/HF, while DG had a significant decrease in RR, SDNN, rMSSD and HF, and significant increase in LF and LF/HF. It was observed that VG had better HRV responses (greater parasympathetic activation) as well as lower levels of perceived stress, while DG had worst HRV responses (greater sympathetic activation) and higher levels of perceived stress. Independently of the environment and activities, individuals’ performance are always affected by their physiological , technicWith the analysis of HRV it is possible to examine the balance of autonomic control , statistical power 1−β = 0.80, and α = 0.05. Given that no study has presented results directly relevant to the effect being targeted in the present study, estimates of effect sizes pertaining to psychophysiological responses to similar experimental manipulations within the context of pre-competitive stress in sports science were used .The sample size was determined through power calculations conducted with G*Power v.3.1 , using ted i.e., . Therefoed i.e., . Based oFifty male eSport players aged 18–29 years  of Brazilian teams took part of the study. The inclusion criteria were being professional athlete with at least 5 years of experience of national or international competitions, and 8–10 h of training daily. The exclusion criteria were to have neuropsychiatric, cardiovascular, or osteoarticular diseases, used any kind of neuropsychiatric drugs, used any caffeinated or alcoholic beverages on the day of the experiment. The study was approved by the Ethics Committee of the Research Center in Sport, Health and Human Development , and all athletes were informed of the inherent risks and benefits of the study before signing an informed consent form.We selected players from 10 different Brazilian teams, all composed of 5 players. If any of the players did not meet the inclusion criteria, the team was not selected to participate in the study. We chose to always carry out the study with two teams per decisive game to facilitate the equalization of the sample to observe the phenomenon of victory and defeat. Thus, after the end of the experiment we had 25 players to Victory Group (VG) and Defeat Group (DG).Players were exposed to decisive games in international championships, such as CS: GO PGL Major Championship Fall and Six Invitational. The experiment was carried out in 2 sessions. In the first one, after signing the informed consent form in the game house, 24 h before the game, anthropometric , physical activity levels through International Physical Activity Questionnaire IPAQ;  and timeThe familiarization procedures with the PSS-10 occurred with the same researcher, 24 h before the experimental conditions, as follows: (1) the researcher read the specific instructions for all questions; (2) the researcher clarified that “there is no right or wrong answer” to the questions and that the answers must be given among the alternatives exposed; (3) the researcher determined for players that there could be no double answers to the same question, and highlighted the importance of veracity in the answers; (4) the researcher asked the players to review their answers before completing the questionnaire. This procedure occurred only during the familiarization phase. In turn, during the experimental conditions the questionnaire was self-reported.As for the HRV collections, the players received instructions on the placement of the chest strap (over the xiphoid process), use of the heart rate monitor and received instructions to remain quiet, with eyes opened, and to breathe spontaneously over the acquisition period.To assess perceived stress, the players responded to the PSS-10. We chose the PSS-10, which is a brief, easy-to-use version with equivalent psychometric properties to the PSS-14, as advised by The HRV data from all players were recorded with players seated on a bench, in an air-conditioned room . After aHRV was recorded at rest using a cardiotachometer Polar RS800cx  at a sampling of 1,000 Hz . A perioThe dependent variables were analyzed in the domain of frequency  and time . The effect size was calculated and then interpreted as suggested by Cohen’s d—0.00 to 0.19 ; 0.20 to 0.49 ; 0.50 to 0.79 (moderate); and ≥ 0.80 . At BL, the assumptions were met for age, weight, height, time of experience, PSS-10, and HRV. Thus, the independent samples t-tests were used to verify the differences between the two groups (VG vs. DG) in BL. A 2 × 2 mixed factor analysis of variance (ANOVA) was used to test for differences between Victory Group vs. Defeat Group (between-group effects) and differences among BL, pre-game, and post-game (within-group effects) for PSS-10, and HRV measurements in the time and frequency domains. Post-hoc analysis was performed using the Bonferroni to assess the effects within each group. The level of significance was determined by 5%  and pre-game (p = 0.541) times, nor intra group difference between BL and pre-game times for VG (p = 0.999) and for DG (p = 0.989). PSS-10 in the post-game time was lower in VG than in DG  = 889.8; p ≤ 0.001], main effects for group , and time  for PSS-10. The interaction revealed a decreased score for PSS-10 in the post-game (4.33 ± 0.96) compared to BL (15.12 ± 1.77) and pre-game (14.75 ± 1.62) times for VG , while also showed an increased score for PSS-10 in the post-game (27.79 ± 1.71) compared to BL (14.70 ± 1.60) and pre-game (14.20 ± 1.84) times for DG .Mixed analysis of variance showed significant group by time interaction , main effects for group [F  = 237.7], and main effects for time [F  = 95.08]. The interaction revealed an increased score of R-R in the post-game (993.44 ± 4.63) compared to BL (950.4± 39.97) and pre-game (951.28 ± 36.15) times for VG , while also showed a decrease in the score of RR in the post-game (749.96 ± 22.46) compared to BL (948.36 ± 37.02) and pre-game (949.52 ± 33.56) times for DG .Concerning time domain measures, for R-R any difference was found between groups in the BL (p = 0.991) and pre-game (p = 0.966) times, nor intra group difference between BL and pre-game times for VG (p = 0.999) and for DG (p = 0.999). However, SDNN in the post-game time was higher in VG than in DG  = 39.82; p ≤ 0.001], main effects for group , and main effects for time . The interaction revealed an increased score of SDNN in the post-game (70.76 ± 3.75) compared to BL (61.6 ± 4.91) and pre-game (61.76 ± 4.9) times for VG , while also showed a decreased score of SDNN in the post-game (57.48 ± 3.36) compared to BL (61.92 ± 4.97) and pre-game (62.28 ± 4.41) times for DG .For SDNN, no difference was found between groups in the BL (p = 0.990) and pre-game (p = 0.899) times, nor intra group difference between BL and pre-game times for VG (p = 0.999) and for DG (p = 0.999). However, rMSSD in the post-game time was higher in VG than in DG  = 50.92; p ≤ 0.001], main effects for group , and main effects for time . The interaction revealed an increased score of rMSSD in the post-game (76.96 ± 2.31) compared to BL (72.96 ± 3.80) and pre-game (72.12 ± 2.18) times for VG , while also showed a decreased score of rMSSD in the post-game (64.4 ± 5.93) compared to BL (73.24 ± 3.46) and pre-game (72.8 ± 3.42) times for DG .For rMSSD, no difference was found between groups in the BL (p = 0.999) and pre-game (p = 0.422) times, nor intra group difference between BL and pre-game times for VG (p = 0.999) and for DG (p = 0.510). In opposition, pNN50 in the post-game time was higher in VG than in DG  = 43.95; p ≤ 0.001], main effects for group , and main effects for time . The interaction revealed an increased score of pNN50 in the post-game (7.04 ± 1.45) compared to BL (3.28 ± 1.13) and pre-game (3.48 ± 0.87) times for VG , while also showed no significant changes in the score of pNN50 in the post-game (3.64 ± 0.86) compared to BL (3.38 ± 1.18) and pre-game (3.92 ± 1.11) times for DG .For pNN50, no difference was found between groups in the BL (p = 0.993) and pre-game (p = 0.999) times, nor intra group difference between BL and pre-game times for VG (p = 0.999) and for DG (p = 0.999). HF in the post-game times was higher in VG than in DG  = 72.24; p ≤ 0.001], main effects for group , and main effects for time . The interaction revealed an increased score of HF in the post-game (8.28 ± 1.20) compared to BL (6.08 ± 1.57) and pre-game (5.96 ± 0.88) times for VG , however with no significant differences between times for while also showed a decreased score of HF in the post-game (3.24 ± 0.72) compared to BL (6.04 ± 1.54) and pre-game (5.88 ± 0.97) times for DG .Considering frequency domain measures, for HF any difference was found between groups in the BL (p = 0.986) and pre-game (p = 0.956) times, nor intra group difference between BL and pre-game times for VG (p = 0.941) and for DG (p = 0.999). However, LF in the post-game time was lower in VG than in DG  = 81.64; p ≤ 0.001], main effects for group  and main effects for time . The interaction revealed a decreased score of LF in the post-game (5.16 ± 0.85) compared to BL (7.72 ± 1.02) and pre-game (8.08 ± 0.86) times for VG , while also showed an increased score of LF in the post-game (9.24 ± 1.09) compared to BL (7.8 ± 0.81) and pre-game (7.96 ± 0.88) times for DG .For LF, no difference was found between groups in the BL (p = 0.870) and pre-game (p = 0.952) times, nor intra group difference between BL and pre-game times for VG (p = 0.999) and for DG (p = 0.999). In opposition, LF-HF in the post-game time was lower in VG than in DG  = 27.55; p ≤ 0.001], and main effects for group . The interaction revealed a decreased score of LF-HF in the post-game (1.42 ± 0.20) compared to BL (2.52 ± 0.78) and pre-game (2.44 ± 0.70) times for VG , while also showed an increased score of LF-HF in the post-game (3.18 ± 0.94) compared to BL (2.41 ± 0.65) and pre-game (2.34 ± 0.66) times for DG .For LF-HF, no difference was found between groups in the BL  in the sinoatrial node of the heart, causing an imbalance between the sympathetic and parasympathetic systems  to control for. As future perspectives, topics such as the use of ergogenic resources to improve neurocognitive and neuromotor performance through caffeine  and braiThe present study investigated the behavior of HRV in the time and frequency domains, and in the perceived stress in different outcomes in eSports athletes during competitive matches. It was observed that VG had better HRV responses (greater parasympathetic activation) as well as lower levels of perceived stress, while DG had worst HRV responses (greater sympathetic activation) and higher levels of perceived stress. Future studies are needed to investigate HRV behavior and emotional states in eSports before and after different outcomes .secm80@gmail.com.The datasets presented in this article are not readily available because the data are under confidentiality requirements. Requests to access the datasets should be directed to SM, The studies involving human participants were reviewed and approved by Ethics Committee of the Research Center in Sport, Health and Human Development . The patients/participants provided their written informed consent to participate in this study.SM, BT, and DM have substantial contributions to the conception or design of the work, drafted the work and revising it critically for important intellectual content provide approval for publication of the content. LO, LC, DT, and FR helped in the acquisition, analysis, and interpretation of data for the work. All authors contributed to the article and approved the submitted version.This research was supported by national funds through the Portuguese Foundation for Science and Technology, I.P., under the project UID04045/2020.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The detection was performed by multiple reaction monitoring (MRM) mode via electrospray ionization (ESI) source in positive and negative ion modes. The product ions m/z 166.1⟶148.1, 285.1⟶123.1, 417.1⟶255.1, and 579.0⟶433.1 were used for determination of ephedrine, gastrodin, liquiritin, and I.S., respectively. The calibration curves of the three analytes were linear with r2 greater than 0.994. The intra and interday precision RSD% was less than 11.5 and 13.4. The intra and interday precision RE% was between −10.4% and 9.33%. The average extraction recoveries of the three analytes were no less than 86.88 ± 1.08%. The developed and validated method was for the first time applied to the pharmacokinetics of three compounds in rat plasma after intragastric administration of Banxia Baizhu Tianma Tang.A rapid and highly selective and sensitive ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC-QQQ-MS/MS) method was applied to simultaneously determine ephedrine, gastrodin, and liquiritin in rat plasma. The three analytes and vitexin-2″-O-rhamnoside (I.S.) were analyzed on a Waters Acquity UPLC C18 column (1.7  Pinellia ternata (Thunb.) Breit. (PT), Atractylodes macrocephala Koidz. (AM), Gastrodia elata Bl. (GE), Poria cocos (Schw.) Wolf (PC), Citrus reticulata Blanco outer peel (CR), and Glycyrrhiza uralensis Fisch. (GU). The formula has the function of drying dampness and resolving phlegm and calming the liver and the wind and is mainly used to treat the wind phlegm dizziness. Modern research reported that the formula has the function of anti-hypertensive and vasodilatory effects [Banxia Baizhu Tianma Tang originates from the book “Yixue Xinwu” of Qing Dynasty, being the classic prescription of traditional Chinese medicine, which consists of six herbs including  effects , and it  effects , 3. Nowa effects , antitum effects  and so oRecently, some literature mainly concentrated on in vitro and in vivo the main chemical composition analyses by HPLC  and UPLCThe six compounds including ephedrine, gastrodin, atractylenolide, pachymic acid, hesperidin, and liquiritin in Banxia Baizhu Tianma Tang, respectively, represent the main chemical markers for the quality evaluation of PT , AM, GE,PT, AM, GE, PC, CR, and GU of Banxia Baizhu Tianma Tang were purchased from the affiliated hospital of Liaoning University of Traditional Chinese Medicine  and authenticated by Prof. Rongxiang Wang, a senior expert of herbal authentication at School of Pharmacy, Liaoning University of Traditional Chinese Medicine. The reference standards including ephedrine, gastrodin, liquiritin, and vitexin-2″-O-rhamnoside   at 30°C was used for the chromatographic separations. The mobile phase consists of 0.1% formic acid aqueous solution (A) and acetonitrile (B), and the gradient elution procedure was 5–35% B from 0 to 5 min, 35%–60 B from 5 to 7 min, 60–90% B from 7 to 9 min, and 90% B from 9 to 11 min. An aliquot of 5 μL was injected, and the flow rate was 0.35 mL/min.An UHPLC system  coupled with an Agilent 6460 Triple Quadrupole Mass Spectrometer  equipped with an electrospray ionization (ESI) source operated in positive and negative ionization modes was used to investigate the pharmacokinetic profiles of ephedrine, gastrodin, and liquiritin in Banxia Baizhu Tianma Tang. A Waters Acquity UPLC C18 column  and collision energy (CE) parameters of multiple reaction monitoring (MRM) to detect the three analytes and I.S. were m/z 166.1 ⟶ 148.1  for ephedrine, m/z 285.1 ⟶ 123.1  for gastrodin, m/z 417.1 ⟶ 255.1  for liquiritin, and m/z 579.0 ⟶ 433.1  for I.S. The other parameters were as follows: drying gas (N2) flow rate, 11.0 L/min; drying gas temperature, 300°C; nebulizer pressure, 15 psig; and capillary voltage, 4000 V.μg/mL in order to prepare calibration curves and quality control (QC) working standard solutions. The final effective concentrations of the calibration standards in rat plasma samples made by spiking 10 μL the corresponding working solutions into 100 μL blank plasma were 2.5, 5, 10, 20, 40, 100, and 250 ng/mL of ephedrine, 50, 100, 200, 400, 1000, and 2500 ng/mL of gastrodin, and 5, 10, 20, 40, 100, 250, and 500 ng/mL of liquiritin. Similarly, the low, middle, and high-concentration QC samples were made by the same way at final concentrations of ephedrine, gastrodin, liquiritin of 5, 40, and 200 ng/mL, 150, 600, and 2000 ng/mL, and 15, 75, and 400 ng/mL, respectively.Stock solutions of calibration standards including ephedrine, gastrodin, liquiritin, and I.S. were, respectively, prepared in methanol at concentrations of 2.40, 1.25, 5.24, and 1.88 mg/mL. The stock solutions of three analytes and I.S. were serially diluted with methanol to be the working standard solutions of 25.0, 10.0, 5.0, and 1.0 μL of plasma sample were spiked with 25 μL of I.S. (0.2 μg/mL) and vortex-mixed for 1 min, and then 300 μL of acetonitrile was added to the sample, vortex-mixed for 1 min, and then centrifuged at 3000 rpm for 15 min. The supernatant was transferred into a 1.5 mL EP tube and evaporated to dryness under 40°C of the stream of nitrogen, and 100 μL of methanol was added, vortex-mixed for 1 min, centrifuged at 15,000 rpm for 3 min, and then transferred to an autosampler vial. An aliquot of 5 μL was injected into the UHPLC-QQQ-MS/MS system.Healthy male SD rats (SCXK2020-0001)  were purchased from Liaoning Changsheng Biotechnology Co. Ltd. . The whole animal experiment process was approved by the Committee of Ethics of Animal Experimentation of Liaoning University of TCM and implemented in strict accordance with the guiding principles of laboratory animal protection. Rats were kept in environmentally controlled breeding room for one week, fed with standard laboratory food as well as water ad libitum, and fasted overnight before the experiments. Aliquots of 100 PT (9 g), AM (18 g), GE (6 g), PC (6 g), CR (6 g), and GU (3 g) in Banxia Baizhu Tianma Tang were, respectively, cut into small pieces, combined and immersed in water for 20–30 min, and then extracted twice with 500 mL water for 1 h and 0.5 h, respectively. The twice extracted solutions were combined, filtered with gauze, and concentrated to about 10 mL under reduced pressure, and then some amount of water was added so that the total volume reached exactly 10 mL that was stored at 4°C before use, and the solution was analyzed by the UHPLC method to obtain the contents of ephedrine, gastrodin, and liquiritin which were 1.699, 0.601, and 0.539 mg/mL, respectively.Selectivity was proved by comparing MRM chromatograms of blank plasma obtained from six rats with those of corresponding standard plasma sample spiked with the three analytes and I.S., and plasma sample after administered to the rats via intragastric administration. The cross-talk of ephedrine, gastrodin, and liquiritin as well as I.S. was evaluated by analyzing upper limit of quantification (ULOQ) plasma samples without I.S. and blank plasma samples only with I.S. The responses of the analyte and I.S. of the low limit of quantification (LLOQ) were, respectively, less than 20% and 5%. Carry-over was assessed by injecting blank samples after injecting ULOQ. The results indicated that the carry-over in blank samples following the ULOQ was not greater than 20% of the analyte response at the LLOQ and 5% of the response for the I.S.c2) least squares linear regression: y = 0.1642x − 0.06984, y = 0.0795x + 0.06003, and y = 0.0051x + 0.20200, with a correlation coefficient (r2) of 0.994, 0.998, and 0.997, respectively, where y is the peak area ratio of the analyte to I.S. and x is the spiked concentration of the analytes.The calibration curves over the concentration range of 2.5, 5, 10, 20, 40, 100, and 250 ng/mL of ephedrine, 50, 100, 200, 400, 1000, and 2500 ng/mL of gastrodin, and 5, 10, 20, 40, 100, 250, and 500 ng/mL of liquiritin were determined using seven or six standard plasma samples via plotting the peak area ratio of the three analytes to I.S. versus the nominal concentration of the analyte in rat plasma. The regression equations of ephedrine, gastrodin, and liquiritin were obtained using weighted (1/The limits of detection (LOD) were determined in signal to noise ratio (S/N) of 3, which were 0.61, 13.7, and 1.36 ng/mL for ephedrine, gastrodin, liquiritin, respectively. LLOQ was defined as the lowest concentration on calibration curve with acceptable precision and accuracy. LLOQs of ephedrine, gastrodin, and liquiritin were 2.5, 50, and 5 ng/mL (S/N > 10) with RSDs of the intra and interday precisions, respectively, below 11.5% and 13.4% and accuracies (RE%) between −10.4% and 9.11%.n = 5), while interday precision and accuracy were determined by repeated analysis on three consecutive days (n = 5 series per day). The RSD and RE should be less than 15%, except at the LLOQ where it should not exceed 20% [Precision and accuracy were evaluated by analyzing the QC samples at low, middle, and high concentrations of the three analytes. Precision was expressed as relative standard deviation (RSD%), and accuracy was expressed as / × 100%. Intraday precision and accuracy were determined by repeated analysis of a set of standards on one day . The matrix effects were investigated at three QC levels by comparing the peak areas of post-extraction blank plasma spiked with the three analytes by directly injecting the pure standard solutions at the same concentrations in six replicates (n = 6). The matrix effect values should be in the range between 85% and 115%. The recovery and matrix effect of the three analytes are listed in The recoveries were performed by comparing the analytical results of extracted samples at three QC levels with corresponding extracts of blank spiked with the analyte after extraction that represented 100% recovery (n = 6). Short-term stability and long-term stability studies were determined at room temperature for 4 h and −20°C for 1 month, respectively. Freeze-thaw stabilities were evaluated after three freeze (−20°C)-thaw (room temperature) cycles. Later, the concentrations obtained after samples were processed and analyzed in this experiment were compared with the nominal values of the QC samples. The stability results are listed in The stabilities of QC samples at three concentrations were studied  being the mobile phase with gradient elution was used to evaluate UHPLC separation and sensitivity in MS detection. Finally, acetonitrile-water containing 0.1% formic acid, which provides better ionization and higher sensitivity, was selected for the separation of the three analytes and I.S., and the excellent peak shape and mass spectral response were also obtained. Three analytes and I.S. were rapidly eluted with total retention times less than 5.0 min and the whole run time was 7.0 min, in which the retention time of ephedrine, gastrodin, I.S., and liquiritin was 1.43, 1.64, 4.45, and 4.85 min, respectively. Typical MRM chromatograms of a blank sample, a blank sample spiked with three analytes and I.S., and a plasma sample at 30 min after intragastric administration in a single dose are shown in Banxia Baizhu Tianma Tang solution of 16.6 mL/kg approximately containing ephedrine of 28 mg, gastrodin of 10 mg, and liquiritin of 9 mg was administered to the rats via intragastric administration. A blood sample of 0.25 mL was withdrawn from orbital venous at times of 0, 5, 10, 20, 30, 45, 60, 90, 120, 180, 240, 360, 480 and 720 min to heparinized polythene tubes and centrifuged at 3000 rpm for 15 min to obtain plasma samples that were stored at −20°C before analysis. The UHPLC-QQQ-MS/MS method described above was validated successfully and applied to the pharmacokinetic studies of ephedrine, gastrodin, and liquiritin in Banxia Baizhu Tianma Tang. Pharmacokinetic analysis was performed using the DAS 2.0 program  to calculate the pharmacokinetic parameters with both compartment and non-compartment approaches. The mean plasma concentration versus time plots of ephedrine, gastrodin, and liquiritin are shown in Cmax) and elimination phase for ISR were selected, which could adequate coverage of the entire PK profile. Therefore, 12 samples  more than 10% of all the 66, 60 and 78 samples of ephedrine, gastrodin and liquiritin were respectively reanalyzed for ISR. Tables 2The samples used to prepare the calibration curves and QC during method validation were not the actual study samples because the differences of protein binding, sample in homogeneity or concomitant medications existed and metabolites, which can affect the accuracy and precision of the analyte in such samples during processing and storage. Therefore, incurred samples were reanalyzed of study samples in separate runs at different days to evaluate accuracy of the samples. The samples around the maximum concentration (c2) gave the best fit according to the Akaike Information Criterion (AIC) values, and the main pharmacokinetic parameters of the three analytes are listed in Cmaxb) of ephedrine, gastrodin, and liquiritin in plasma were, respectively, 128.2 ± 28.8, 1622.7 ± 657.3, and 244.7 ± 64.2 ng/mL, along with the corresponding values of the area under the concentration-time curves (AUCt0⟶a) of the three analytes were 262.2 ± 42.2, 2483.5 ± 1194.0, and 230.3 ± 135.5 ng h/mL, meaning that ephedrine and liquiritin had a little weak absorption compared with gastrodin. Moreover, both apparent volume of distribution (Vda) and clearance (CLa) of ephedrine and liquiritin were larger than those of gastrodin, indicating that ephedrine and liquiritin were mainly distributed in surrounding tissues leading to lower plasma concentrations. In addition, the peak time (Tmaxb) of ephedrine, gastrodin, and liquiritin was 0.917, 0.500, and 0.167 h, respectively, suggesting that the three analytes were all rapidly absorbed, in which the liquiritin was the fastest one. The terminal half-life (t1/2b) 2.639, 1.278, and 2.176 h, indicated that the eliminations of them were slow. Furthermore, it was noteworthy that ephedrine and liquiritin could be detected up to 720 min, gastrodin up to 240 min after dosed, meaning that they exert a longer lasting effect in vivo in rats, and the advantage of TCM was that the chemical compositions can remain a longer time in body and exert a lasting therapeutic effect and increase the clinical efficacy.In the study, pharmacokinetic analysis was performed using the DAS 2.0 program to calculate the pharmacokinetic parameters with both compartment (a) and non-compartment (b) approaches, a two-compartment open model to gastrodin, and one-compartment open model to ephedrine and liquiritin (weight = 1/A developed and validated UHPLC-QQQ-MS/MS method was successfully applied to the pharmacokinetic studies of ephedrine, gastrodin, and liquiritin in rat plasma. Three components in Banxia Baizhu Tianma Tang are simultaneously determined in rat plasma after intragastric administration for the first time. This experiment provides a pharmacokinetic basis for Banxia Baizhu Tianma Tang in clinical aspects and can make Banxia Baizhu Tianma Tang better applied in clinical treatment. Although the pharmacokinetic study of traditional Chinese medicine is very difficult due to its very low content of chemical components in each single herb, ephedrine, gastrodin, and liquiritin which are representative of the herbs of PT, GE, and GU in Banxia Baizhu Tianma Tang can be determined in the study, and further studies will be conducted on other components in this formula in the future.


There has been considerable interest in the interrelationship between the liver and hypertension. The relationship between serum total bile acid (TBA) and hypertension has been reported. Moreover, intrahepatic cholestasis of pregnancy was correlated to gestation hypertension. However, the association between maternal serum TBA level in the normal range and new-onset hypertension disorders during pregnancy remains unclear. The present study aimed to evaluate the relationship between maternal serum TBA level in the normal range and the risk, disease severity and adverse pregnancy outcomes of new-onset hypertension during pregnancy.Using the electronic medical records on all pregnant women from the Department of Obstetrics and Gynecology, Third Affiliated Hospital of Guangzhou Medical University, between 2014 and 2020, we conducted a retrospective cohort study of 2581 singleton pregnant women with maternal serum TBA levels in the normal range. Patients were grouped into the non-hypertension during pregnancy (1071), gestational hypertension (480) and preeclampsia (1030) groups.p < 0.01). Multiple logistic regression analysis showed that TBA level was independently and significantly associated with preeclampsia and gestational hypertension . Moreover, elevated TBA level was positively associated with the risk of severe PE and negatively with mild PE (p < 0.01). In addition, maternal serum TBA levels were negatively related to birth weight (p < 0.001).We found that maternal serum TBA levels were significantly higher in the preeclampsia and gestational hypertension groups than in the non-hypertension group (These results suggest that maternal serum TBA in the normal range also might be a valuable biomarker for disease severity in preeclampsia and gestational hypertension. Additionally, our results also indicate associations of serum total bile acid levels in the normal range with an increased risk of fetal growth restriction and low birth weight among offspring. These results suggest that TBA could serve as a prognostic biomarker for new-onset hypertension during pregnancy. Hypertensive disorders of pregnancy (HDP) are common gestational complications that cause 10–15% of maternal deaths every year worldwide . Among tRecently, there has been more interest in the interrelationship between liver dysfunction and hypertension. The liver, the center of metabolism, has complicated functions, including synthesis and secretion of proteins, biotransformation of biomolecules, carbohydrate metabolism, lipid metabolism and bile acid metabolism . NumerouThe synthesis and secretion of bile acid (BA) is one of the most important functions of the liver. BA has been found to be a part of the digestive system over the last few decades and is important for chyle digestion and absorption of lipid-soluble vitamins , 10. Hown = 1030) and GH (n = 480) subgroups.As shown in Fig. @8000 modular analyzer series , and T bilirubin, ALB and 24-h proteinuria Roche cobas@ C501.Maternal clinical parameters were collected from medical record, including maternal age, gestational age at delivery (GA), body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP), whether receive in-vitro fertilization (IVF), nulliparity, pregestational diabetes infant birth weight and body length. SBP and DBP data in HDP group were collected for the first time measured after diagnosis, and in the non-HDP group, SBP and DBP were collected for the first time measured after hospitalized. Maternal fasting blood samples were collected by first-time examination after hospitalization, after an overnight fast of at least 8 h at a mean gestational age of 20 to 34 weeks. The biomarkers included TBA, ALB, TC, TG, AST, ALT, T bilirubin, and 24-h proteinuria. TBA, TC, TG, AST, ALT were assessed by Roche cobasThe criteria suggested by the American College of Obstetricians and Gynecologists (ACOG) for the diagnosis of pregnancy-associated hypertension were adopted. PE was defined as systolic blood pressure ≥ 140 mmHg or diastolic blood pressure ≥ 90 mmHg over at least two intervals of 6 h after 20 weeks of gestation with previously normal blood pressure and new-onset proteinuria (> 0.3 g/24-h or at least > 1 + on protein dipstick when urine could not be collected for 24 h). And liver function impaired, thrombocytopenia, severe persistent right upper quadrant or epigastric pain and not accounted for by alternative diagnoses, renal insufficiency, pulmonary edema, new onset headache unresponsive to acetaminophen and visual disturbances should diagnosed as PE for women with gestational hypertension in the absence of proteinuria .The primary pregnancy outcomes in our study were low birth weight and fetal growth restriction (FGR). FGR was diagnosed following the guidelines for FGR , as indiData were presented as the mean ± standard deviation or median (25–75 percentile). According to the normality of the variable distributions, measurement data were expressed as the means ± standard deviations, except for GA, SBP, DBP, TBA, AST, ALT, 24 h proteinuria and birth length. An independent-samples t-test was performed for age, BMI, ALB, TC, TG and birth weight. Count data were expressed as proportions (%), and the Mann–Whitney U test was performed for GA, SBP, DBP, TBA, AST, ALT, 24 h proteinuria and birth length. Spearman correlation analysis was used to evaluate the correlation between maternal serum TBA levels and birth weight and between AST/ALT and TBA levels. Multiple logistic regression analysis was performed to examine the influence of the following variables: maternal age, prepregnancy BMI, AST, ALT, T bilirubin, TC, TG, ALB, pregestational diabetes, nulliparity and IVF. The odds ratios with 95% CIs were calculated.p values < 0.05 were considered indicative of statistical significance.All statistical analyses were performed with IBM SPSS Statistics version 20 software  and GraphPad Prism 6 ; P < 0.01) and an earlier gestational age at delivery than the non-HDP patients (interquartile range (IQR) of GA: 39 (38–40) vs. 38 (37–39), 37 (34–38), for non-HDP, GH and PE, respectively). Similar to other reports, HDP patients tended to have a higher BMI (P < 0.01), AST and ALT levels (p < 0.01). TC and TG levels were significantly higher in the PE group (p < 0.01) than in the non-HDP group but not in the GH group. Smoking and drinking were considered as risk factors for HDP [After ineligible patients were excluded, 2581 pregnant women were included for subsequent analysis. 480 women were diagnosed with GH, 1030 were diagnosed with PE, and 1071 pregnant women were non-HDP. The characteristics of the pregnant women among these groups were shown in Table  for HDP ; howeverρ = 0.8, p < 0.01, ρ = 1.58, p < 0.01, respectively, data not shown) in pregnant women, which is consistent with a previous report [In our study, the mean serum level of TBA was 2.2 (1.5–3.4) µmol/L for the GH patients and 2.1 (1.5–3.2) µmol/L for the PE patients, which was approximately 1.3-fold higher than that in the non-HDP group (1.8 (1.18–2.7) µmol/L) ; beyond that, TG was also an independent factor associated with PE and GH .Multivariate logistic regression analysis was used to test whether serum TBA was significantly and independently correlated with PE and GH Fig. . As in pr = 0.2096, p < 0.0001), DBP  and 24 h-proteinuria .The diagnosis of PE was based on blood pressure and proteinuria. Thus, we asked if maternal serum TBA level was associated with systolic blood pressure, diastolic blood pressure and 24 h-proteinuria. We apply Spearman correlation analysis between serum TBA and SBP, DBP, 24 h-proteinuria in patients that diagnosed for HDP. As shown in Fig. Next, we tested the relationship between serum TBA and the severity of PE. Based on a previous report , we catePreeclampsia is divided into mild preeclampsia and sever preeclampsia according to the severity. Among PE patients, increased serum TBA levels showed more percentage of severe PE. TBA levels between 1.6 to 4.1 µmol/L , higher than 4.1 µmol/L and less than 8.2 µmol/L  and between 8.2 µmol/L and 10 µmol/L  had higher risks of severe PE. And for mild PE, TBA levels between 1.6 to 4.1 µmol/L , higher than 4.1 µmol/L and less than 8.2 µmol/L  and between 8.2 µmol/L and 10 µmol/L   and GA  , fetal growth restriction (FGR), and earlier GA. Thus, we performed Spearman correlation to test the relationship of maternal serum TBA and GA as well as birth weight in HDP patients. The results showed that TBA was significantly correlated with birth weight  and between 8.2 µmol/L and 10 µmol/L  carry a higher risk for LBW. Pregnancies with serum TBA levels higher than 4.1 µmol/L and less than 8.2 µmol/L  and between 8.2 µmol/L and 10 µmol/L  had a higher risk for FGR (Table This study had the following findings: 1) Maternal serum TBA was significantly and positively associated with HDP and severity of PE; 2) The TBA level was negatively related to birth weight and FGR. Taken together, these findings indicate that maternal serum TBA is a potential prognostic biomarker of PE.Intrahepatic cholestasis of pregnancy (ICP) was determined by pruritus occurring during pregnancy and serum TBA above 10 µmol/L, and excluding for elevated serum aminotransferase and/or serum bile acids related to other causes of liver test abnormalities , 25. ICPIn our study, we demonstrate that maternal serum TBA levels in the normal range were significantly higher in the HDP group than those in the non-HDP group and could be an independent risk factor for HDP. In addition, the level of serum TBA is significantly and positively associated with the severity of PE. Although, total bile acid was not significant change during pregnancy, especially second and third trimesters , 30. HigFetal complications, especially LBW and FGR, have been reported in PE and ICP pregnancies. A recent study in China reported increased TBA levels with a decrease in birth weight . Song anTo our knowledge, this is the first study to investigate the relationship between HDP and maternal serum TBA levels in the normal range. In addition, we included hospitalized patients, which ensured the integrity of the information and allowed us to study the relationship between HDP and TBA. However, the present study has some limitations. First, this study is a single-center study. Second, because this study is a cross-sectional observational study, causality cannot be determined. Third, although blood sample were collected after overnight fasting, we do not measure hormone level, dietary habit and emotional status that are factors that can influence the TBA level. Last, bile acid has many secondary forms; in this study, we discuss only the total bile acid in serum.In conclusion, the present study demonstrates that maternal serum TBA can be an indicator of new-onset HDP, especially PE. Moreover, an increasing level of TBA can be a cost-effective predictor for PE, which is associated with both clinical severity and pregnancy outcomes. Therefore, maternal serum TBA has the potential to be a biomarker for PE and to facilitate disease management.


It is important to reorient the acetabular fragment into an optimal position and version to ensure a good long-term outcome after Bernese periacetabular osteotomy (PAO). Unfortunately, the intraoperative balance between overcorrection and undercorrection remains challenging for the surgeon. The purpose of this study was to answer two questions: (1) Does the femoral head coverage measured on intraoperative fluoroscopy agree with that measured on postoperative radiography? (2) What is the reliability of intraoperative fluoroscopy in identifying hip center correction in PAO?p value of < 0.05. Bland–Altman analysis, conducted using GraphPad Software (version 9), was used to quantify the agreement between intraoperative fluoroscopic images and postoperative radiographs.A total of 173 patients (173 hips) who underwent PAO for developmental dysplasia of the hip (DDH) at our center from July 01, 2020, to December 31, 2020, were retrospectively reviewed. Imaging data from 111 patients  were included in this study. The analysis included measurement of the lateral center-edge angle (LCEA), acetabular index (AI), anterior wall index (AWI), posterior wall index (PWI), extrusion index (EI), and medial offset distance (MO). These measurements were acquired from intraoperative fluoroscopic images and postoperative radiographs and compared by paired t test using SPSS (version 24.0). Significance was determined at a p = 0.90, 0.95, and 0.83, respectively), but those in the AI, PWI, and MO were significant (p < 0.05). The mean biases (95% limits of agreement) of the LCEA, AI, AWI, PWI, EI, and MO were − 0.04 (− 6.85), − 0.97 (− 7.78), 0 (− 0.30), − 0.11 (− 0.36), − 0.12 (− 11.92), and − 1.11 (− 5.51), respectively.The means  of the LCEA, AI, AWI, PWI, EI, and MO obtained on intraoperative fluoroscopy were 32.86° (5.73°), 0.66° (5.55), 0.29 (0.10), 0.75 (0.17), 11.15% (6.50%), and 8.49 mm (3.68 mm), respectively. On postoperative radiography, the corresponding values were 32.91° (6.31°), 1.63° (5.22°), 0.29 (0.15), 0.85 (0.14), 11.27% (7.36%), and 9.60 mm (3.79 mm). The differences in the LCEA, AWI, and EI acquired from intraoperative fluoroscopic images and postoperative radiographs were not significant (The LCEA, EI, and AWI can be used to reliably predict postoperative femoral head coverage at the level of 2D graphics. Acetabular inclination can be cautiously assessed using AI on intraoperative fluoroscopy. In the absence of intraoperative 3D image evaluation, the AWI and PWI demonstrate acceptable agreement between fluoroscopy and radiography in assessing the acetabular version. Although the MO shows slight bias, it can be helpful in properly positioning the acetabulum during PAO. Bernese periacetabular osteotomy (PAO), introduced by Ganz in 1988 –5. HowevAfter the osteotomy, the acetabulum can be freely rotated and mobilized, which not only improves the femoral coverage but also optimizes the rotation center of the hip joint. Although the anatomy around the acetabulum is complex and close to the iliac vessels and sciatic nerves, it is not very difficult for surgeons to complete the osteotomy with monitoring by intraoperative fluoroscopy after clearing the learning curve. However, reorienting and confirming the optimal orientation and version of the acetabular fragment , as wellIntraoperative radiography or fluoroscopy, computer-assisted navigation , customiThis study was performed with the aim of answering two questions: (1) Does the femoral head coverage measured on intraoperative fluoroscopy agree with that measured on postoperative radiography? (2) What is the reliability of intraoperative fluoroscopy in identifying hip center correction in PAO?After receiving institutional ethics review board approval, we retrospectively reviewed the radiographs of all 173 patients (173 hips) who underwent PAO for developmental dysplasia of the hip (DDH) at our center between July 01, 2020, and December 31, 2020. As previously described , Bernes All patients’ preoperative computed tomography (CT) images of the pelvis, images obtained by intraoperative fluoroscopy, and postoperative standing pelvic AP radiographs were obtained from our hospital’s picture archiving and communication system (PACS). The CT scanning parameters were as follows: 120 kV; 300 mAs; matrix, 512 × 512; pitch, 0.7539; field of view (FOV), 300–400 mm; and slice thickness, 0.75 mm. To eliminate pelvic tilt and rotation during surgery, we referred to Lehmann’s method [All standing AP radiographs of the pelvis and intraoperative fluoroscopic images were obtained as per the protocols recommended in previous studies , 12, 15.Intraoperative fluoroscopic images and postoperative standing AP radiographs of the pelvis were analyzed by an orthopedic surgeon (J.P.). The analysis included measurement of the LCEA, AI, anterior wall index (AWI), posterior wall index (PWI), extrusion index (EI), and medial offset distance (MO), as described in previous reports –19. The p value of < 0.05. Bland–Altman analysis, conducted using GraphPad Software (version 9), was used to quantify the agreement between intraoperative fluoroscopy and postoperative radiography. The bias was estimated by calculating the mean difference and the 95% limits of agreement (LOA) between the intraoperative fluoroscopic images and the postoperative radiographs.The LCEA, AI, AWI, PWI, EI, and MO acquired by intraoperative fluoroscopy and postoperative radiography were compared by paired t test using SPSS (version 24.0). Significance was determined at a The results of the paired t test and the Bland–Altman analysis are shown in Table p = 0.90, 0.95, and 0.83, respectively); however, there was a significant difference in the AI, PWI, and MO between intraoperative fluoroscopy and postoperative radiography (p < 0.05).The means  of the LCEA, AI, AWI, PWI, EI, and MO obtained on intraoperative fluoroscopy were 32.86° (5.73°), 0.66° (5.55), 0.29 (0.10), 0.75 (0.17), 11.15% (6.50%), and 8.49 mm (3.68 mm), respectively. The corresponding parameters obtained on postoperative radiography were 32.91° (6.31°), 1.63° (5.22°), 0.29 (0.15°), 0.85 (0.14°), 11.27% (7.36°), and 9.60 mm (3.79 mm), respectively. According to the paired t test, the LCEA, AWI, and EI determined using the two imaging modalities were not significantly different . Since multiple prior studies have reported on interrater and intrarater reliability in measuring the LCEA, AI, AWI, PWI, EI, and MO, we did not repeat such assessments. Third, we determined that the bias of the AI, PWI, and MO as measured on fluoroscopy was acceptable based on the reference normal values reported in previous studies. Further studies are needed to determine whether the bias of these parameters affects hip function after PAO. Fourth, we did not investigate the radiograph in the supine position after the operation. However, the stability of the hip joint in the weight-bearing position is more important regarding the occurrence of pathological hip changes and symptoms. Fifth, we did not use intraoperative 3D images to evaluate femoral head coverage. Intraoperative 3D evaluation may be a more accurate method. However, this is a relatively expensive method that has not been widely used in China. Since 3D images can more accurately evaluate the coverage of the acetabulum to the femoral head, we will add postoperative 3D image data in future studies to further evaluate the accuracy and reliability of intraoperative fluoroscopy.p < 0.05). We suspect that this difference may be due to the difficulty in determining the medial margin of the acetabular sourcil on fluoroscopic images. Charles [The LCEA of Wiberg, AI, and EI was used to assess the lateral coverage of the femoral head. Correction of the LCEA between 25° and 40°, AI between 0° and 10°, and EI ≤ 20% were defined as the target ranges after PAO based on previously published normative values , 23. Cha Charles  also conProper acetabular reorientation includes not only lateral but also anterior and posterior coverage. Excessive anterior coverage is a detriment to posterior coverage and may cause impingement and adversely affect the long-term survival of the joint after PAO . An anteA lateralized hip center is considered to be a sign of structural instability. The hip center is considered lateralized if the medial aspect of the femoral head is greater than 10 mm from the ilioischial line . MedialiThe LCEA, EI, and AWI can reliably predict postoperative femoral head coverage at the level of 2D graphics. The use of AI in the intraoperative assessment of acetabular inclination requires caution. In the absence of intraoperative 3D image evaluation, the agreement in the AWI and PWI between the two imaging modalities is acceptable in assessing the acetabular version. We acknowledge that it is difficult to assess the hip center position intraoperatively. Although the MO demonstrates slight bias, it can nevertheless help position the acetabulum properly during PAO.