The mean IOP elevation was 34.41 ± 9.70 mmHg in the whole study population, and the difference in IOP elevation between OAG and nonglaucoma subgroups was not significant (p = 0.159). In the whole group, the presuction IOP, postdilated pupil size (PPS), and central corneal thickness (CCT) were positively corrected to higher IOP elevation , while axial length (AL) was negatively related to IOP elevation . For the OAG subgroup, the longer AL was more significantly correlated to lower IOP elevation compared to those without glaucoma . In conclusion, presuction IOP, PPS, and CCT are related to higher IOP during FLAC, while the AL is negatively correlated to the IOP elevation in FLACS, especially for patients with OAG. Reviewing these parameters before FLACS may enable physicians to find patients who are at risk of IOP elevation.In this study, we evaluate the association between biometrics and intraocular pressure (IOP) during femtosecond laser-assisted cataract surgery (FLACS) in normal patients and those with open-angle glaucoma (OAG). A retrospective cross-sectional study was conducted. A total of 103 patients who had received elective FLACS were enrolled, and those with OAG who received FLACS were further divided into a subgroup. The perioperative IOP of FLACS was measured before, during, and after the suction procedure. Demographic data and preoperative biometrics were collected from the medical records. The generalized linear model was applied to yield the adjusted odds ratio (aOR) and corresponding 95% confidence interval (CI) of each biometric for the IOP elevation in the whole group and the OAG subgroup. The mean preoperative IOP was 20.96 ± 4.79 mmHg, which rose to 55.37 ± 11.58 mmHg during suction, and decreased to 23.75 ± 6.42 mmHg after suction; the IOP both during and after suction was significantly higher than the presuction IOP (both  Femtosecond laser-assisted cataract surgery (FLACS), first introduced in 2008, is widely applied for patients who need cataract surgery to improve visual acuity . CornealFor the mechanism of IOP elevation in FLACS, stabilizing the eye during FLACS with fixation devices is a necessary step, such as with laser in situ keratomileusis, and may lead to the transient elevation of IOP ,8. The cRegarding the correlation between elevated IOP and other ocular parameters in cataract surgery, the mean preoperative IOP and deep anterior chamber depth (ACD) were correlated to the postoperative IOP in conventional cataract surgery , and canThe aim of our study is to evaluate the potential predictive factors, mainly ocular biometrics, for increasing intraoperative IOP in patients who have received FLACS using the fluid-filled interface. In addition, the potential different effect of ocular biometrics on IOP elevation during FLACS between OAG and nonglaucoma patients was analyzed.A retrospective cross-sectional study was conducted in the Taipei Municipal Wanfang Hospital. Patients scheduled for elective FLACS and intraocular lens implantation who were over 20 years old were enrolled in the study population. The exclusion criteria were: (1) angle-closure glaucoma or uncontrolled OAG with an IOP more than 21 mmHg despite medical treatment; (2) active inflammation in the eye; (3) a history of prominent corneal disease or corneal surgery, except superficial corneal injury; (4) a CCT less than 500 mm or greater than 600 mm; (5) active iris neovascularization; (6) the presence of intraocular silicone oil; or (7) ocular or systemic steroid use three months before the preoperative visit. After the selection process, a total of 103 patients who had undergone FLCS were included. In the study population, 12 patients were diagnosed with OAG and constituted the OAG subgroup in the statistical analysis.® Vision System, Alcon, Fort Worth, TX, USA) in all patients. Directly after surgery, patients were asked whether they had vision problems during the procedure, and symptoms were recorded if they had occurred. Topical levofloxacin eyedrops and prednisolone emulsion were prescribed postoperatively four times per day for one week.All FLACS procedures were performed in one eye of each patient by two well-trained surgeons (Chien-Liang Wu and Yi-Zhen He) with the same femtosecond laser system . The patient interface of LensAR was composed of two parts: a suction ring that contacted only the sclera with an outer diameter of 18.0 mm and a disposable lens that rigidly mated the suction ring to the surgical system. In the first step, the suction ring  is centr® XL , which was covered by individually wrapped Ocu-Film and tip covers before every application; then, the calibration was checked. Pre- and post-treatment IOP measurements were performed three times in each patient, and the average value was used, in which the time interval between the removal of the ring the post-treatment IOP measurement was within five seconds for all participants. For the IOP during the suction procedure, records were extracted from the FLACS device by the same surgeon (either Yi-Zhen He or Chien-Liang Wu). In addition, total suction time (ST) was collected from the same FLACS device. The above data were applied in subsequent analyses.Demographic data and medical history were obtained from the medical records. Preoperative biometrics, including ACD, CCT, and axial length (AL) were obtained by Optical Biometer AL-Scan . Postdilated pupil size (PPS) was recorded after dilatation using the FLACS device. Regarding the IOP measurement, pre- and post-treatment IOPs were measured by another physician (Ya-Hui Wang) using a Tono-Penp value less than 0.05 was considered to indicate statistical significance, and a p value less than 0.001 was depicted as p < 0.001.SPSS version 20.0  was utilized for statistical analysis in our study, except for power calculation. The statistical power of the OAG subgroup reached 0.73 under a 0.05 alpha value and medium effect size using G*power version 3.1.9.2 . Descriptive analysis was used to present the demography, systemic disease, existence of OAG, biometrics, ST, and IOP values, in which numerical indices are shown as the mean ± standard deviation (SD). Repeated one-way analysis of variance was used to compare the IOP value before, during, and after the FLACS procedure, and we constructed a box plot to demonstrate the distribution of IOP in each measurement. Then, the amount of IOP elevation was calculated via the following formula: IOP during FLACS minus the presuction IOP. In the next step, the generalized linear model was utilized to produce the adjusted odds ratio (aOR) and corresponding 95% confidence interval (CI) of the presuction IOP, PPS, and all the biometrics of the IOP elevation, which incorporated the effects of age, sex, eye laterality, systemic disease, the presence of OAG, and all ocular parameters. For subgroup analysis, the Mann–Whitney U test was used to compare the IOP value between the OAG and nonglaucoma subgroups. Furthermore, the generalized linear model was applied again to reveal the aOR and 95% CI of ocular parameters for IOP elevation in patients with OAG compared to nonglaucoma participants. For analysis with three or more values, we used Bonferroni correction/adjustment to refine the statistical results of multiple comparisons. A The mean age of the study population was 66 ± 12.60 years old, and there were 52 male and 51 female patients in the study population. A total of 58 patients received FLACS for the right eye, and another 45 subjects received FLACS for the left eye. The details of systemic disease numbers and preoperative biometrics are shown in p < 0.001). After the removal of the suction ring, the mean IOP reduced to 23.75 ± 6.42 mmHg, which was significantly lower than the IOP during suction (p < 0.001), but still higher than that in the preoperative status (p < 0.001) (p = 0.137), peak IOP , and IOP elevation  compared to nonglaucoma patients, while the postsuction IOP was significantly higher in the OAG subgroup .The mean preoperative IOP was 20.96 ± 4.79 mmHg and rose to 55.37 ± 11.58 mmHg upon application of the suction ring and vacuum with significant difference (< 0.001) . In addip = 0.038), larger PPS , and thicker CCT  were positively correlated to higher IOP elevation in the multivariable model. On the other hand, longer AL was correlated to lower IOP elevation  after adjusting for all indices (p = 0.006). The effects of other parameters were similar between the two subgroups , and the effect of each parameter on IOP elevation in the two subgroups was similar to that in the whole group (Regarding the correlation between ocular parameters and IOP elevation, higher presuction IOP (aOR: 2.399, 95%CI: 1.770–8.521,  indices . In subgle group .Briefly, our study demonstrates that an IOP elevation would occur after the performance of FLACS with a fluid-filled interface. The AL was negatively correlated to IOP elevation during FLACS, while presuction IOP, CCT, and PPS were positively related to IOP elevation. Moreover, the negative correlation between longer AL and IOP elevation was more prominent in OAG individuals compared to that in nonglaucoma patients.Regarding the potential predicting factors for IOP elevation during FLACS, there was a relatively rare finding. In the study written by Liu et al., and Chang et al., both preoperative IOP and ACD were positively correlated to a higher postoperative IOP in patients with angle-closure glaucoma who had received traditional cataract surgery ,14. In aRegarding other ocular parameters and the elevated IOP, presuction IOP, PPS, and CCT showed positively significant associations with higher IOP elevation in the current study. The effect of presuction IOP on the IOP elevation was established in previous studies ,16, and Research to evaluate the influence of ocular parameters on IOP elevation during FLACS in OAG individuals is rare. In the current study, the longer AL was associated with lower IOP elevation in OAG patients compared to nonglaucoma patients, and all the parameters showed a similar effect in the whole group, OAG subgroup, and nonglaucoma subgroup. In a previous study, OAG patients with an AL shorter than 26 mm were associated with larger 24-h IOP fluctuation . No patiPatients with glaucoma would experience a higher elevation of IOP during FLACS compared to nonglaucomatous individuals, while the long-term effect of such IOP elevation is not fully elucidated . In our For the interface device in FLACS, a flat applanating contact lens was first introduced for femtosecond laser flap creation . This inThere are still limitations in our study. First, the absence of a control group made the comparison of different basic characters difficult. Second, we used tonopen only for the IOP measurement, while the application of a Pascal tonometer should be considered in subsequent research. Patient numbers between the OAG subgroups and the nonglaucoma subgroups were extremely uneven due to the retrospective design and could contribute to some statistical bias. The statistical power of the OAG subgroup was also relatively inadequate, yielding a power of 0.73. However, we also presented the effect of ocular parameters on IOP elevation in the general population; thus, we did not reduce the nonglaucoma participants, and recruiting some recent OAG cases who received FLACS may increase the heterogeneity of our study population concerning the time of surgery.In conclusion, AL is negatively correlated to IOP elevation in patients receiving FLACS, especially for those with OAG. Furthermore, presuction IOP, PPS, and CCT are positively associated with IOP elevation in FLACS. Consequently, repeated examination of these parameters may be recommended for patients scheduled for FLACS, and the use of FLACS should be considered carefully in OAG patients with short AL. Further prospective study to quantify the influence of FLACS with a fluid-filled interface on different severities of glaucoma is mandatory.


The treatment of choice for patients with advanced heart failure (HF) and with limiting symptoms with evidence of a poor prognosis despite optimal conventional treatment is a heart transplant. However, there is little literature dealing with the effects of cardiovascular prehabilitation with an important change in physical capacity, which can influence the admission on the waiting list for a heart transplant. We presented one young male, smoker, with no prior history of cardiovascular disease, severe ventricular dysfunction, interventricular defect, and HF. It was decided to implant a cardioverter-defibrillator as primary prevention of sudden death and start the pre-cardiac transplant evaluation and subsequent inclusion in the waiting list on an elective basis. While waiting for the transplant, cardiopulmonary rehabilitation (CPR) was indicated. After 15 months of CPR, the patient improved his left ventricular ejection fraction , systolic pulmonary artery pressure (55 to 40 mmHg), and peak oxygen uptake (23.9 to 29.1 ml/kg/min). In this patient, a program of CPR improved cardiac function and physical capacity, allowing him to be removed from the national waiting list for a heart transplant. The treatment of choice for patients with advanced heart failure (HF) and with limiting symptoms with evidence of a poor prognosis despite optimal conventional treatment is heart transplant . Unfortu2peak) of <10 mL/kg/min is a strong predictor of poor prognosis [2peak lower than 50% can also be used to guide transplant candidacy in conjunction with absolute value VO2peak [Through the cardiopulmonary exercise testing (CPET), the objective assessment of physical capacity provides prognostic information. In patients, achieving a peak oxygen uptake VOpeak of < VO2peak .Exercise training led to an increase in multiple systemic, angiogenetic, and platelet-derived inflammatory mediators . AdditioThe literature suggests a multifactorial beneficial effect of cardiopulmonary rehabilitation (CPR) after a heart transplant, improving autonomic control, muscle strength, and body composition -10, but Prehabilitation programs have shown efficacy in increasing functional capacity and preventing postoperative complications in some high-risk surgical populations . NeverthIn this context, there is little literature dealing with the effects of cardiovascular prehabilitation with an important change in physical capacity, which can influence the admission on the waiting list for a heart transplant.A 42-year-old male, office employee, smoker, height 1.66 m, weight 66 kg, with no family history or antecedents of cardiovascular disease, who consulted at Instituto Cardiovascular de Rosario (ICR) for a condition characterized by progressive dyspnea lasting approximately two months, associated with episodes of mid-thoracic pain with radiation to the shoulders two hours long.In the initial evaluation, signs and symptoms compatible with HF and holosystolic murmur in a polifocal bar were evidenced, with the presence of thrill and gallop rhythm. He was admitted and further evaluated by: (i) routine biochemistry examination within normal parameters except for an elevated N-terminal pro-brain natriuretic peptide (NT-ProBNP) of 5321 pg/ml. (ii) 12-lead electrocardiogram revealed sinus tachycardia of 130 beats per minute associated with signs of inferior-lateral myocardial fibrosis with compromised right leads. (iii) Chest X-ray, where signs of grade III venulo-capillary hypertension were observed. (iv) Color Doppler echocardiogram: systolic and diastolic diameter of the left ventricle 61/55 mm, respectively, impaired left ventricular ejection fraction , abnormal segmental motility with apical akinesia in all its segments, inferior, septal, and anteromedial. Systolic displacement of the plane of the tricuspid annulus was 13 mm and grade II diastolic dysfunction. Pulmonary artery systolic pressure was 55 mmHg. At the junction of the middle and apical third of the interventricular septum, a continuity solution with a left to right flow of approximately 5.5 mm was observed , clopidogrel 75 mg daily (8 hours), bisoprolol 2.5 mg every 12 hours (8-20 hours), ivabradine 7.5 mg every 12 hours (8-20 hours), enalapril 5 mg every 12 hours (8-20 hours), eplerenone 50 mg per day (8 hours), furosemide 20 mg per day (8 hours), atorvastatin 80 mg per day (20 hours) with good clinical response.Once his admission condition had stabilized, post-infarction risk stratification was continued by: (i) Cinecoronariography: the left main coronary artery did not present lesions, while angiographically significant stenotic lesions were evidenced (>60%) in the circumflex, anterior descending, and right coronary artery. (ii) Cardiac magnetic resonance imaging (MRI): severely increased ventricular volumes and severely depressed LVEF (28%). At the septoapical level, a 3.5 mm muscular interventricular communication was visualized. Absence of myocardial viability.Due to the high risk of the patient due to severe ventricular dysfunction, interventricular defect, and HF, it was decided to implant a cardioverter-defibrillator as primary prevention of sudden death and to start the pre-cardiac transplant evaluation and subsequent inclusion on the waiting list on an elective basis. At the time of his discharge, outpatient CPR was indicated.On January 31, 2020, the patient entered the CPR program. The sessions lasted 60 minutes, where the first 30 minutes were for aerobic training and the remaining 30 minutes for exercises for muscle strength, balance, coordination, and flexibility. ECG telemetric control and pulse oximetry were used to monitor the patient during the first sessions. The patient had a functional class III on the New York Heart Association (NYHA) scale at admission to the program. The first five weeks were for adaptation to training, where the intensity was lower than 4 (0-10) on the Borg scale.2peak of 23.9 mL/kg/min, finding his anaerobic threshold at 100 beats per minute (bpm) and 70% of the VO2peak reached, and a maximum heart rate toward the end of the test of 124 bpm. Concerning the theoretical maximum heart rate (TMHR), it reached 70%, although the patient was on beta-blocker medication. The hemodynamic response of the patient during the test was normal, without presenting chest pain. The respiratory exchange ratio (RER) was 1.09 (Table The first cardiopulmonary exercise test (CPET) was performed on a treadmill on March 6, 2020. The protocol used was modified Bruce, where the intensity was increasing every three minutes. The reason for stopping the test was due to exhaustion and the request of the patient in stage 4 of Bruce protocol. At that time, the patient reached a VOWith these results, starting at week 5, the treadmill aerobic training was planned at an intensity of 80% with a variable continuous training method, where progressively, and depending on the patient's responses during the sessions, we modified the training protocol to perform four minutes at 80% interspersed by two minutes of recovery at 40%. Concerning the strength exercises, 4 or 5 exercises were performed per session, with an intensity measured by the Borg scale that should not be higher than level 5 when reaching the last repetitions of each series. As in aerobic training, depending on the patient's responses and adaptations, the number of series, number of repetitions, and complexity of the exercises progressed with the running of the sessions. The training protocol was carried out based on the recommendations of the American Association of Cardiovascular and Pulmonary Rehabilitation, the American Heart Association, and the American College of Cardiology guidelines .2peak of 29.1 mL/kg/min, finding his anaerobic threshold at 125 bpm and 86% of VO2peak and a maximum heart rate of 136 bpm (82% of his TMHR). The RER was 1.09.After 15 months within the CPR program and 122 sessions carried out, with a frequency of two-weekly sessions, a new follow-up CPET with the same protocol was carried out on May 14, 2021. It is essential to clarify that the patient was with the same medication as the baseline, although, at that time, he had NYHA functional class 0 and gained 3 kg. The test was stopped in stage 4 at the request of the patient. At the end of the test, the patient reached a VOA second color Doppler echocardiogram was performed in May 2021, finding: left ventricular systolic and diastolic diameter of 54/41 mm, respectively, LVEF with moderate deterioration of 40%, abnormal segmental motility with apical, inferior and inferoseptal akinesia. Systolic displacement of the plane of the tricuspid annulus of 22 mm. Pulmonary artery systolic pressure of 40 mmHg. Interventricular communication of approximately 5.2 mm.2peak value was reached during the second CPET and the absence of hospitalizations was associated with HF in this period, his cardiologist decided to exclude him from the national waiting list for a heart transplant and to continue within the CPR program, to maintain and/or improve his current physical condition.Due to the clinical improvements of the patient, the VOHF harms the health-related quality of life (HRQoL), both in physical function and psychosocial function, worsening the deterioration with increased functional class . In thisCPR has been widely demonstrated to improve physical fitness, functional capacity, and HRQoL in chronic HF patients . Even moIn this case, the exercise was before the transplant, a concept defined as prehabilitation. A recent study evaluated a multimodal prehabilitation intervention in HF patients awaiting heart transplant showing that CPR is feasible, safe and may prevent the deterioration in exercise, functional capacity, and quality of life while waiting for a transplant .2peak is considered the best predictor of survival in cardiovascular diseases [2peak. In addition, VO2peak values have a critical role in informing patient selection for advanced HF interventions such as heart transplants [The VOdiseases . CPET prnsplants . Previous studies have indicated that a peak aerobic power ≤10 mL/kg/min is a strong predictor of a poor prognosis in patients with HF . In addi2peak improved from 23.9 to 29.1 ml/kg/min, and more important was the improvement in the appearance of the anaerobic threshold, which would suggest that the patient was less fatigued and more resistant to the aerobic effort, due to the rightward deviation of the threshold. One possible explanation is physiological and clinical adaptations to training, particularly peripheral muscle adaptations that contribute to the improvements shown in the CPET [In this present case, the exercise capacity analyzed through the measurement of VOFurther studies will be needed to show the effects of the prehabilitation in this population and to suggest a CPR guideline in patients with severe HF awaiting a heart transplant.As shown in this case report, the authors were able to carry out safe and effective CPR, which allowed the patient to significantly improve their quality of life and exercise capacity and get off the cardiac transplant list for clinical stability. Therefore, CPR can be a good strategy in patients who are candidates for a heart transplant, particularly in many developing countries, where referral of patients to a heart transplant list for presurgical CPR is not frequent.


Mesenchymal stem cells (MSCs) have become promising candidates for regeneration medicine due to their multidifferentiation potential and immunomodulatory ability. Compared with classic MSCs derived from the bone marrow and fat, dental-derived MSCs show high plasticity, accessibility, and applicability. Therefore, they are considered alternative sources for regeneration medicine.  Four types of MSCs were isolated from the dental pulp, periodontal ligament, dental follicle, and alveolar bone of the same donor, and there were five different individuals. We analyzed their morphology, immunophenotype, proliferation rate, apoptosis, trilineage differentiation potential, and the gene expression during osteogenic differentiation.  Our research demonstrated that DPSCs, PDLSCs, DFPCs and ABMMSCs exhibited similar morphology and immunophenotype. DFPCs showed a higher rate of proliferation and apoptosis. When cultured in the trilineage differentiation medium, all types of MSCs presented the differentiation potential of osteogenesis, adipogenesis, and chondrogenesis. Through staining and genetic analysis during osteogenic induction, ABMMSCs and PDLSCs showed the highest osteogenic ability, followed by DPSCs, and DFPCs were the lowest.  Overall, our results indicated that different dental-derived stem cells possessed different biological characteristics. For bone tissue engineering, ABMMSCs and PDLSCs can be used as optimal candidates of seed cells. Bone regeneration is a challenging issue in clinical treatment. A variety of causes such as trauma, congenital abnormalities, or tumor resection, can destroy the bone and cause dysfunction. To date, autologous tissue transplantation provides a possible treatment method with the development of functional reconstruction surgery . HoweverMesenchymal stem cells (MSCs) are pluripotent progenitor cells that can differentiate into different kinds of lineages, including osteoblasts, adipocytes, chondrocytes, epithelial cells, neuron-like cells, and hepatocyte-like cells . These cSo far, a variety of MSCs from dental tissues have been isolated and characterized, including dental pulp stem cells (DPSCs) , periodoIn this study, we replaced traditional BMMSCs with ABMMSCs that can be obtained with low trauma and systematically compared the morphology, immunophenotype, proliferation rate, antiapoptosis ability, and trilineage differentiation potential of DPSCs, PDLSCs, DFPCs, and ABMMSCs. The osteogenic ability and gene expression patterns in the osteogenic process of four types of stem cells were compared to evaluate their clinical application potential in repairing bone defect in order to provide theoretical support for the selection of the best seed cells of bone tissue engineering.3. Then, they were digested with 3 mg/mL type I collagenase  for 40 minutes at 37°C. After that, the tissues were passed through a 70 μm strainer ; then, the suspension was centrifuged at 1200 rpm for 5 minutes. The cell pellets were resuspended in MEM-α medium  containing 20% fetal bovine serum  and 100 U/mL of penicillin-streptomycin  and then were plated into dishes. For the isolation of ABMMSCs, the alveolar bone was crushed into small pieces and seeded in dishes using the same medium. All the types of cells were incubated at 37°C in a humidified atmosphere with 5% CO2. At around 80% confluence, cells were detached by 0.25% trypsin  and split at the ratio of 1 : 3. The medium for all types of cells was refreshed every 3 days. Cells at passage 3 were utilized for further experiments.The healthy teeth and alveolar bone samples came from the extracted third molar, and the donors were 16-20 years old. Each type of MSC coming from the same donor was used for the following experiments, and there were five different individuals. All the procedures were approved by the Research Ethics Committee of Shanghai Jiao Tong University School of Medicine (SH9H-2020-TK60-1). For the isolation of DPSCs, PDLSCs, and DFPCs, the dental pulp tissue, dental follicle tissue, and one-third of the periodontal ligament tissue on the root surface were washed with phosphate-buffered saline (PBS) and cut into small pieces of 1-2 mmThe four types of MSCs at passage 3 were harvested by trypin-EDTA  and centrifuged at 1200 rpm for 5 minutes. Then, the cell pellets were resuspended in PBS and transferred to the test pipe. The immunophenotype was analyzed with the following antibodies: FITC-conjugated CD24, CD29, CD44, CD45, and CD73 and PE-conjugated CD34, CD90, CD105, and CD146. Corresponding isotype-matched antibodies were used as controls. All the antibodies were purchased from BD Bioscience . Cells were incubated in the dark for 30 minutes and collected into a flow cytometer. The results were analyzed by using Cell-Quest for Macintosh Software.MSCs were cultured on cell slides until reaching 80% confluence and then replaced with serum-free medium for 24 hours. Then, they were fixed with 4% PFA for 30 minutes and washed 3 times with PBS. Then, we followed the manufacturer's instructions of TUNEL detection kit  and took pictures under a fluorescence microscope. The red light represented the apoptotic cell nucleus stained by TUNEL, and the blue light was the cells stained by DAPI. The percentage of the number of red fluorescent cells to the number of blue fluorescent cells represented the proportion of apoptotic cells in the total cells.3 cells/well, and each well contained 100 μL α-MEM with 10% FBS. After the incubation for the first 24 h, the cell number was tested every 24 h for seven consecutive days. To determine the number of MSCs, 10 μL of CCK-8 reagent was added to each well and then incubated for 2 hours at 37°C. The optical density value at a wavelength of 450 nm was detected with Multiskan GO . Each experiment was performed in three replicates.According to the manufacturer's instructions, Cell Counting Kit-8  was used to detect cell proliferation. The four types of MSCs were seeded in 96-well plates  at a density of 3 × 105 cells/well and maintained in MEM-α medium containing 10% FBS. When cells reached 100% confluency, the medium was replaced with the adipogenic differentiation medium  according to the manufacturer's instructions and kept for 21 days. Lipid droplets of the resultant differentiated cells were detected using Oil red staining .For adipogenic differentiation, the four types of MSCs were seeded in each well of 6-well plates at the density of 2 × 105 cells were centrifuged in 15-mL polypropylene tube  and resuspended in 500 μL chondrogenic differentiation medium . The medium was refreshed every 3 days. After 21 days, the cell pellets were fixed in 4% PFA and then cut into paraffin sections of 5 μm thickness. To assess the deposition of glycosaminoglycans, the sections were stained with alcian blue .For chondrogenic differentiation, 5 × 105 cells/well and maintained in MEM-α medium containing 10% FBS. When the cells reached 60-70% confluence, the medium was replaced with the osteogenic differentiation medium . Cells were then seeded in osteogenic differentiation medium for 21 days, and the medium was refreshed every 3 days.For osteogenic differentiation, the MSCs were seeded in each well of 6-well plates at the density of 1 × 10After osteogenic differentiation, the MSCs were detected by ALP staining kit  and ALP activity assay kit  at days 3, 7, and 14. The cells were incubated in ALP staining solution for 20 minutes at 37°C. A scanner  was used to capture digital images. ALP activity was normalized by the protein concentration. Following standard protocols, the proteins were extracted using RIPA lysis buffer  and concentrations were measured by Bradford protein assay kit .After osteogenic differentiation for 21 days, the MSCs were detected for mineralized nodules by using alizarin red staining solution . The cells were fixed in 4% PFA for 30 minutes and incubated with alizarin red staining solution at room temperature. After 5 minutes, the reactions were then stopped by washing with PBS. A scanner and microscope was used to capture digital images. After staining with alizarin red, the calcium nodules were dissolved by 10% CPC and incubated at 37°C for 30 minutes. Then, the supernatant was transferred to a 96-well plate, and the absorbance at 590 nm was measured for statistical analysis.ΔΔCt- values obtained by normalization to β-actin RNA as the internal control.Total RNA was extracted by using RNAiso Plus  during trilineage differentiation of the MSCs. RNA concentration and purity were measured by using a Multiskan GO . The cDNA was synthesized from RNA by using a retrotranscription kit . Quantitative real-time polymerase chain reaction (qRT-PCR) was performed using SYBR Green I master mix  and running in a LightCycler 96 . Primer sequences are outlined in α medium containing 10% FBS were used as a control. The samples were heated for 8 min at 100°C. Equal amount of proteins was separated by 10% SDS-polyacrylamide gel and transferred to polyvinylidene difluoride (PVDF) membranes . The membranes were blocked with 5% skim milk for 2 hours at room temperature and then incubated with primary antibodies RUNX2 , ALP , OSX , OPN , and β-actin  overnight at 4°C. After washing with TBST buffer, the blots were incubated with HRP goat anti-rabbit  for 1 hour at room temperature. The proteins were visualized using a ChemiDoc imaging system .After 21 days of osteogenic differentiation of DPSCs, PDLSCs, DFPCs, and ABMMSCs, the total cellular protein was extracted using the RIPA lysis buffer , and concentrations were measured by Bradford protein assay kit . MSCs that were cultured in MEM-p value; p < 0.05 was considered statistically significant .All statistical analyses were calculated using GraphPad Prism 8 Software. Differences among more than three groups, one-way ANOVA was applied. All experiments were repeated three times at least, and the data are presented as mean ± SD. Results were considered statistically significant with After isolating and cultivating DPSCs, PDLSCs, DFPCs and ABMMSCs, the MSCs were expanded and passaged in the culture medium. An inverted phase-contrast microscope was used to evaluate the morphology of the MSCs at passages 0 and 3 . We founWe analyzed the immunophenotype of MSCs by flow cytometry. The data showed a low expression of endothelial and hematopoietic markers  and high expression of typical MSC markers  and CD29, CD44, and CD146 . The resWe induced the MSC apoptosis through serum deprivation. The results showed that the apoptotic rate of DFPCs was the highest and that of ABMMSCs was the lowest, followed by DPSCs and PDLSCs. There was no statistical difference in the apoptotic rate between DPSCs and PDLSCs Figures .According to the CCK-8 test results for consecutive 7 days, the growth curve was analyzed and drawn. The results showed that all the MSCs proliferated slowly in the first two days and entered the logarithmic phase at the third day. The proliferation rate of DFPCs was significantly faster than that of ABMMSCs, and DPSCs and PDLSCs were the second .ALP is mainly expressed in the mineralized matrix produced by osteogenic differentiated cells and is considered to be an early marker of osteoblast . In the In order to detect the osteogenic capacity of DPSCs, PDLSCs, DFPCs, and ABMMSCs, we performed osteogenic differentiation experiments. Each type of cells was cultured in osteogenic differentiation medium for 21 days. The staining results confirmed that all the MSCs had undergone osteogenic differentiation, but the efficiency was different. As shown in Figures We further evaluated the osteogenic capacity of MSCs by comparing the relative mRNA expression of osteogenic markers at days 0, 7, 14, and 21 of osteogenic induction . Except We also examined the expression of osteogenic differentiation protein markers RUNX2, ALP, OSX, and OPN using western blotting Figures . After 2In order to test the adipogenic capacity of DPSCs, PDLSC, DFPCs, and ABMMSCs, the MSCs were cultured in the adipogenic induction medium. At about 7 days after induction, the MSCs became flat and lipid vacuoles began to appear in the induced cells. After 21 days of induction, the cells were fixed by 4% PFA and then the adipogenesis was verified by oil red O staining. The accumulation of cytoplasmic oil droplets could be clearly observed in the four types of MSCs . We furtFor the chondrogenic capacity of DPSCs, PDLSC, DFPCs, and ABMMSCs, the MSCs were centrifuged in a 15 mL conical tube. After culturing with the cartilage differentiation medium for 21 days, the chondrocyte pellets were fixed and cut into paraffin sections. The sections were then stained with alcian blue, which indicated chondrogenic glycosides. All tested MSCs showed positive staining results . We furtMSCs play a key role in the balance between health and disease . By combThe evaluation of the best seed cells for tissue engineering includes many aspects. In addition to considering the characteristics of cells which contain proliferation and differentiating into specific lineages, it is also important to determine the source of the stem cells. Compared with the two somatic MSCs which are BMMSCs and ADSCs, dental-derived stem cells are becoming a promising cell source of regenerative medicine because the tissues can be obtained from medical waste such as wisdom teeth, which is simple and less moral controversy. At the same time, these organizations can be stored for use. For BMMSCs, additional invasive procedures may cause serious complications of the donor, such as infection. In addition, the lower cell mass can be obtained is a disadvantage encountered when isolating BMMSCs. Although ADSCs can be obtained by liposuction surgery or caesarean section, the progress is still invasive , 34. In In order to avoid individual differences, we isolated four types of cells from the same individual. Through the identification of cell surface markers and the ability of tri-lineage differentiation, the cells we isolated meet the standards of MSCs . For celThe antiapoptosis ability of stem cells is also an important aspect when considering therapeutic applications. It has been reported that about 99% of MSCs are lost in the first 24 hours after cell transplantation, and apoptosis is considered to be the main factor . A lot oIn addition, we studied the osteogenic differentiation capacity of DPSCs, PDLSCs, DFPCs, and ABMMSCs. Osteogenic differentiation ability was confirmed by ALP staining, ALP activity assay, and alizarin red staining. After osteogenic induction, ABMMSCs showed more ALP staining at day 7 and darker at day 14, followed by DPSCs and PDLSCs, and DFPCs possessed the lightest color. The ALP activity test results were consistent with the ALP staining. At day 21 of osteogenic induction, ABMMSCs showed stronger alizarin red staining, PDLSCs was second but slightly stronger than DPSCs, and DFPCs was the lightest. These results indicated that ABMMSCs produced more mineralized nodules in the process of osteogenic induction and were most sensitive to the differentiation of osteoblasts.On the genetic level, we analyzed the expression of osteogenic markers  to further prove osteogenic differentiation. The results showed that as the osteogenic induction time increased, the expression of osteogenic genes in ABMMSCs increased significantly and higher than DPSCs, PDLSCs, and DFPCs. At the same time, we analyzed the protein level at day 21 with or without osteogenic induction. At the late stage of osteogenic induction, the MSCs still show osteogenic potential. The expression of osteogenic proteins in ABMMSCs was significantly higher than that of DFPCs, followed by PDLSCs and DPSCs. In summary, our results showed that compared with the abovementioned four types of dental-derived stem cells, ABMMSCs possessed the strongest osteogenic capacity, PDLSCs were slightly weaker, DPSCs were second, and they were all better than DFPCs. Considering the availability of cell sources, proliferation, apoptosis, and osteogenic ability, ABMMSCs and PDLSCs are considered to be potential substitutes for BMMSCs in bone tissue engineering.In summary, our results indicated that DPSCs, PDLSCs, DFPCs, and ABMMSCs from the same individual possessed different biological characteristics. ABMMSCs and PDLSCs showed a higher osteogenic potential, which would make them the optimal stem cells for bone tissue engineering. However, we have only compared the biological characteristics and osteogenic potential of these stem cells in vitro. More research needs to be done in vivo to characterize the optimal stem cells source for bone regeneration.


Dietary intervention included dietary assessment, individual feedback, and dietary guidance. The dietary intakes between the two groups were compared using the Mann-Whitney U test. At 6 months postpartum, the energy-adjusted intakes of protein, total dietary fiber, potassium, magnesium, phosphorus, zinc, vitamin B6, and β-carotene were significantly higher in the intervention group than in the control group. The changes in energy-adjusted intakes of total dietary fiber and iron from 1 month postpartum to 6 months postpartum were significantly different between the two groups. Dietary intervention based on the health belief model improved nutrition at 6 months postpartum, although the impact was limited.No dietary intervention that focuses on the diet quality of postpartum women has been developed in Japan, although most postpartum women experience an insufficient intake of vitamins and minerals. We aimed to examine whether dietary intervention, based on the health belief model, at both 1 and 3 months postpartum affects nutrient intake and food group consumption at 6 months postpartum. A randomized controlled trial was conducted at a university hospital in Tokyo between 2015 and 2016. Healthy women at 1 month postpartum were randomly allocated to either an intervention group ( Adequate nutrition is essential during the months following childbirth because the mother is recovering from childbirth and any nutrients consumed by the mother pass to the baby through breast milk . NeverthThe majority of postpartum women in Japan do not receive any dietary guidance since individual dietary assessment is not a component of the hospital stay after childbirth and standard medical checkups during the postpartum period . NeverthIn Western countries, several dietary education interventions for weight loss and diabetes mellitus in the postpartum period have been developed ,8,9,10. The present study aimed to examine whether dietary intervention, including individual dietary assessment and feedback and providing dietary information for women at 1 and 3 months postpartum, affects the intake of nutrients and food groups at 6 months postpartum.A randomized control trial was conducted in a university hospital in Tokyo, Japan, between June 2015 and July 2016. Participants were allocated 1:1 to either an intervention group (IG) or a control group (CG) using concealed random allocation from a computer-generated random numbers table. The research secretary managing the table followed a simple up-to-down method for participant allocation and provided the researchers with the respective intervention allocation and serial ID number at the time of recruitment. This study did not blind the participants due to the fact that it was a behavioral guidance intervention trial; thus, the participants knew which group they were allocated to.Healthy Japanese women were recruited at their 1-month postpartum medical checkup. In Japan, the medical checkups are routinely performed. Thus, almost all women visit a hospital at 1 month postpartum. Women were excluded if they were less than 20 years of age, had inadequate Japanese literacy, had diabetes, hypertension, or psychiatric diseases, followed a restricted diet due to certain diseases, or had a baby with a serious health condition.The dietary intervention was developed based on the HBM ,13,14. AThe CG received their individual dietary results of the three investigations  and the above-mentioned leaflet by mail after the investigation at 6 months postpartum. Participants in the CG had usual care, including a home visit by public health nurses, midwives, or childcare advisors. Home visits to families with infants are required by law; thus, were conducted in both CG and IG. During home visits, mothers are often educated on dietary intake for breastfeeding and preventing anemia and constipation .The participants were asked to complete a diet history questionnaire and an original questionnaire including demographic questions within 7 days of recruitment. Missing or unclear data were resolved through telephone interviews. The medical charts of the participants were reviewed to obtain information on their pregnancies and childbirths. At 3 and 6 months postpartum, participants were asked to answer another mail-in questionnaire.Primary outcomes were nutrient intake and food group consumption at 6 months postpartum, and eventual changes in nutrient intake and food group consumption between 1 and 6 months postpartum. The secondary outcome was changes in body weight at 6 months postpartum relative to the measurement at 1 month postpartum.6, vitamin B12, folate, vitamin C, and β-carotene) were calculated using an ad hoc computer algorithm for the BDHQ. A validation study reported correlation coefficients of 0.26 to 0.63 between the BDHQ and semi-weighed 16-day dietary records among Japanese women for these nutrient intakes [The original questionnaire distributed to the mothers at 1 month postpartum contained questions on age, parity, education levels, household income, height, smoking habits, regular supplement usage, and infant feeding style. Maternal pre-pregnancy weight, maternal weight at 1 month postpartum, and birth weight of the infants were obtained from medical charts. Dietary intake was assessed using a brief-type self-administered diet history questionnaire (BDHQ). The BDHQ comprised a four-page fixed-portion questionnaire that assessed habitual dietary intake in the previous month and included a total of 58 food and beverage items ,18. Stan intakes ,18. We cInformation on dietary intake, weight, and infant feeding style was collected at 3 and 6 months postpartum using questionnaires.This study was conducted according to the guidelines of the Declaration of Helsinki. The Ethics Committee of the university approved the study procedures and protocol . The study was registered with the University Hospital Medical Information Network-Clinical trial registration. All of the participants provided informed written consent prior to the baseline investigation.The sample size was calculated with a focus on iron intake, since it is one of the most significant nutrients in the postpartum period. We estimated a total sample size of 208 women for an expected effect size of 0.40, a power of 0.80, and an alpha of 0.05, based on iron intake reported in a previous study . These sp-value < 0.05 were considered statistically significant. Statistical analyses were performed using the Statistical Package for Social Sciences version 24.0 .Student’s t-test, Chi-square test, or Fisher’s exact test were used to compare the baseline characteristics of the participants between the IG and CG. Mann-Whitney U tests were employed to compare the nutrient intake and food group consumption between the two groups at 1, 3, and 6 months postpartum, and to assess potential changes in these variables during the course of the study. All differences with a two-sided n = 149) or the CG (n = 147). Of these women, 33 did not submit questionnaires at the baseline investigation  and 3 women returned incomplete data. Thus, 260 women  provided complete baseline data and were asked to answer further questionnaires. Thirty-six women were excluded at 3 months postpartum , and a further 30 women were excluded at 6 months postpartum . Thus, 100 women in the IG and 94 women in the CG completed questionnaires at 6 months postpartum. No significant differences in demographic variables were found between the dropout subjects and the participants who completed all investigations (data not shown).Of the 442 healthy women who were eligible at 1 month postpartum, 296 (67.0%) provided written informed consent . Particip = 0.039). No significant differences in nutrient intake and food group consumption were found between lactating and non-lactating women (data not shown).p = 0.033; data not shown).At 3 months postpartum, only the energy-adjusted zinc intake was significantly higher in the IG than in the CG (p = 0.036), total dietary fiber (p = 0.022), potassium (p = 0.049), magnesium (p = 0.046), phosphorus (p = 0.024), zinc (p = 0.020), vitamin B6 (p = 0.022), and β-carotene (p = 0.047) were significantly higher in the IG than in the CG at 6 months postpartum. Changes in energy-adjusted intakes of total dietary fiber (p = 0.039) and iron (p = 0.040) from 1 to 6 months postpartum were significantly different between the two groups.At 6 months postpartum, 88 women (88.0%) reported any types of breastfeeding, and 12 (12.0%) reported formula feeding in the IG. In the CG, 84 women (89.4%) reported any types of breastfeeding, and 10 (10.6%) reported formula feeding. No difference was found in breastfeeding rates between the two groups.p = 0.002) (p = 0.001).Confectionery consumption was significantly lower in the IG than in the CG at 6 months postpartum (= 0.002) . In addiThe mean body weight (standard deviation) was 53.4 (6.5) kg in the IG and 53.3 (7.2) kg in the CG at 3 months postpartum, and 52.7 (7.2) kg in the IG and 52.6 (7.3) kg in the CG at 6 months postpartum. There were no significant differences in the weight change from baseline to 3 or 6 months postpartum between the two groups.6, and β-carotene. Our results suggest that approaches to each perception in the HBM improved dietary behaviors during the postpartum period. In addition, second dietary feedback at 3 months postpartum might be effective as a cue to action. This was speculated from the results, showing that the intake of several nutrients at 6 months postpartum were significantly different between the two groups, despite the lack of clear differences in nutrient intake at 3 months postpartum. A dietary intervention pilot study in the United States was also conducted between 2 and 6 months postpartum and was successful in increasing intakes of dark green and deep yellow vegetables [Dietary intervention based on HBM had positive effects on intake of the following nutrients in postpartum Japanese women: protein, total dietary fiber, potassium, magnesium, phosphorus, iron, zinc, vitamin Bgetables . The posgetables ,22,23. Hgetables ,25, and getables . Thus, dThe energy-adjusted iron intake at 6 months postpartum increased in more than 50% of women in the IG compared to the assessment at 1 month postpartum, while it decreased in the CG. Maternal hemoglobin recovery is often delayed, especially in lactating women with low iron intake, because the iron levels in breast milk are stable irrespective of maternal dietary intake . However6, and β-carotene at 6 months postpartum were higher in the IG than in the CG. Although there was no significant difference in the energy intake between the two groups, significant differences in several energy-adjusted nutrient intakes would indicate an improved quality of diet. Protein was a target nutrient in this intervention because it is essential for the development of breastfed infants [6, and β-carotene are also essential for physical recovery from childbirth and maintaining an adequate supply of these nutrients in breast milk. In addition, dietary fiber is important for the prevention of constipation. Although explanations of these nutrients were not included in the intervention leaflet, the nutrients were contained in other nutrient-rich foods and recommended food groups that were introduced in the leaflet. For instance, dietary fiber is contained in a wide range of foods, such as pulses, potatoes, vegetables, fruits, and seaweeds, all of which were recommended in the leaflet. Thus, the nutrient intake might have increased as a secondary effect by consciously eating foods recommended in the leaflet. It is also possible that the leaflet generated an interest in a healthy diet in the IG, leading them to seek further information on other important nutrients and change their dietary intake. In the prenatal Japanese study [Energy-adjusted intakes of protein, total dietary fiber, potassium, magnesium, phosphorus, zinc, vitamin B infants . Proteinse study , no nutrIn terms of food groups, participants in the IG consumed less confectionery at 6 months postpartum. Another randomized controlled trial for postpartum women who were overweight and obese has also shown a reduction in confectionery consumption . Our intNo significant differences in changes in body weight and energy intake were found between the two groups, and the mean weights at 6 months postpartum in both groups were similar to those pre-pregnancy. Energy intake was far below the estimated energy requirement  in most participants, even after considering potential dietary under-reporting . JapanesThe CG received only usual care in the present study. This might have made it difficult to interpret the results of the intervention, that is, to distinguish the effects of the intervention itself from the effects of the attention received by the intervention. However, we speculate that two dietary investigations at 1 and 3 months postpartum may have prompted some participants in the CG to become interested in dietary intake. In addition, all of the families with infants in Japan had home visits by public health nurses or midwives, as usual care. Home visits often include dietary guidance for mothers . This opOur study has several limitations. Firstly, the participant characteristics may have been biased because the research was conducted at a single university hospital in an urban area. The mean age of the participants in this study was slightly higher than that of the participants in national reports . Such di6, and β-carotene at 6 months postpartum. Such dietary intervention would help to improve dietary intake of postpartum women in clinical settings, although the observed effects were comparatively limited. Further intervention research, which adds other components, such as increasing self-efficacy in changing dietary habits, is required to enhance the effects.This randomized controlled trial initiated in women at 1 month postpartum indicates beneficial effects of individual dietary intervention based on HBM on the intake of protein, total dietary fiber, potassium, magnesium, phosphorus, iron, zinc, vitamin B


The epicardium is an essential cell population during cardiac development. It contributes different cell types to the developing heart through epithelial-to-mesenchymal transition (EMT) and it secretes paracrine factors that support cardiac tissue formation. In the adult heart the epicardium is a quiescent layer of cells which can be reactivated upon ischemic injury, initiating an embryonic-like response in the epicardium that contributes to post-injury repair processes. Therefore, the epicardial layer is considered an interesting target population to stimulate endogenous repair mechanisms. To date it is still not clear whether there are distinct cell populations in the epicardium that contribute to specific lineages or aid in cardiac repair, or that the epicardium functions as a whole. To address this putative heterogeneity, novel techniques such as single cell RNA sequencing (scRNA seq) are being applied. In this review, we summarize the role of the epicardium during development and after injury and provide an overview of the most recent insights into the cellular composition and diversity of the epicardium. Ischemic heart disease, and especially myocardial infarction (MI) remains a major cause of death globally . MI is pThe epicardium is a single-cell layer of mesothelial origin located on the outside of the heart. Intriguingly, this cell type is of crucial importance during cardiac development. In brief, the epicardium contributes cardiac cell types to the developing heart , 14, it In the healthy adult heart, the epicardium is a quiescent layer. However, it is reactivated after certain types of injury and subsequently it recapitulates several of its developmental processes. In animal models that display the potential for cardiac regeneration such as zebrafish and neonatal mammals , 22, theTo optimize the post-injury response, it is important to understand the processes underlying the activation of the epicardium and the regulation of its differentiation into cardiac cell types. An unresolved question in this context has been whether the whole epicardial population can participate, or whether distinct cell types reside within the epicardial layer that have specific abilities within the reparative response. With the advent of single cell sequencing, we are gaining more insight into the composition and the potential contribution of endogenous cells in the heart. Here, we will highlight the role of the epicardium during development and cardiac repair and discuss novel insights on the composition of this cell layer based on single cell RNA sequencing (scRNA seq) data.via the formation of free-floating cell aggregates or via direct contact with the myocardium . The PEO is an evolutionary conserved cluster of cells that develops from the lateral plate mesoderm and is located at the venous pole of the heart near the septum transversum. In mice, the PEO becomes visible around embryonic day 8.5 (E8.5) , a stageocardium –30, whilronectin , 32. Uporonectin , 34.Identification of the PEO as a transient structure has relied on scanning electron microscopy (SEM)  and staiThe vital role of the epicardium for cardiac development was highlighted by studies in an avian model where epicardial outgrowth from the PEO was physically inhibited. This led to the formation of a thin myocardium and malformation of the coronary vasculature, amongst other developmental defects , 40, indWt1, Tbx18, Tcf21, GATA5, Scx, and Sema3D have been used to trace the fate of epicardial cells based on transgene expression. An even better controlled lineage trace system can be achieved by fusing Cre to a mutated ligand-binding domain of the human estrogen receptor, in which recombination relies on the presence of tamoxifen. This provides lineage tracing with a temporal control  arises as well as through recruitment of extracardiac progenitors epicardium  is driven by promoters that are considered specific to epicardial cells. By crossing these mice with transgenic reporter lines containing a lox-flanked stop-codon followed by a reporter gene, cell specific Cre expression results in indefinite expression of a reporter protein like Green Fluorescent Protein (GFP) or β-galactosidase. Several promoters of epicardial related genes such as  control , as demoebrafish , 54, 55.ebrafish , 56–58. genitors . HoweverCre mice . Others nary ECs . Interesicardium . Neverthicardium . Additiovia expression of epithelial cadherin , which is responsible for maintaining cell-cell adhesion and adherens junctions. The EMT-inducing transcription factors, including Snail Family Transcriptional Repressors 1 and 2 (SNAI1/2), zinc finger E-box binding family members 1 and 2 (ZEB1/2), and twist-related protein 1 (TWIST1)   can all (TWIST1) . Wt1 is (TWIST1) , 61, 62.(TWIST1) , 64. Howdial EMT . This subleeding , 62. Addm of WT1 . Indeed,m of WT1 .In vitro, using mouse primary epicardial cells, a bi-directional role for WT1 and Tbx18 was reported. Wt1 knockdown induced epicardial EMT through expression of Snai2, which could be reversed by knockdown of Tbx18, thus acting as a regulator of EMT , a class II basic helix-loop-helix (bHLH) transcription factor, is another epicardially expressed protein that is also involved in the regulation of EMT and the differentiation into various cellular lineages. Depletion of Tcf21 during early stages in henotype . In Tcf2ntiation . The regntiation . These sex vivo heart culture model, deletion of Mrtfa and Mrtfb reduced EPDC migration. Additionally, in embryos lacking both Mrtfa and Mrtfb, epicardial integrity was compromised, as well as the coronary angiogenesis due to reduced epicardium-derived pericytes , nuclear factor of activated T-cells 1 (NFATC1) and protein arginine methyltransferase 1 (PRMT1). The MRTF serum-response factor (SRF) regulatory network modulates epicardial migration and invasion. Cell motility is driven by interactions between SRF and MRTF-A/B, modulating the expression of regulators of actin dynamics . In an eericytes . NFATC1,ericytes , is exprericytes . It was ynthesis . HoweverSnai2 in epicardial EMT in mice. Interestingly, the reduction of p53 levels in Tbx18-mediated PRMT1 knockout mice normalized the disrupted invasion, as well as the formation of epicardium-derived mesenchymal lineages  family and its receptors (FGFR). For example, FGFR1 is expressed both in the PEO and in the epicardium, and loss of FGFR1 in quail embryos was shown to reduce the myocardial invasion of epicardial cells . In contaffected , indicatvia receptor splice variants FGFR1c and FGFR2c  produced by the epicardium and promotes proliferation and differentiation of cardiomyocytes d FGFR2c . Additiod FGFR2c , 78. Besnd Tcf21 , two TFsvia IGF2 . RA alsoignaling . Conversa FGFR2b .Wt1-KO mice have deficient epicardial expression of angiogenic factors Vegfa and Angpt1, suggesting a contribution to abnormal coronary vessel development (via C-X-C motif receptor 4 (CXCR4) on nearby endothelial cells epicardial development , 91. TGFdial EMT . Moreoveof EPDCs –97. Overof EPDCs .Tbx18 and Raldh2, are merely expressed at low levels, indicating that the epicardium is in an inactive state in the healthy adult heart  in ECs  in ECs , which c) in ECs , indicatWt1 and Raldh2 by binding to their enhancer elements  family of transcription factors was identified as a regulator of elements . More repression .Cre lineage-trace model, cells derived from the epicardium were reported to contribute to fibroblast, EC and CM lineages , a peptide secreted by endothelial cells and the epicardium during development and after injury  to isolate these cells, they found that the EPDCs secrete pro-angiogenic factors in vitro. Further analysis revealed that FGF2 and VEGFA were in large part responsible for these effects , a factor present within the secretome of adult rat epicardial cells nd swine . Interesocardium . This fiocardium . Neverthocardium , was injocardium . Similar lineage . In both lineage . In a mor injury . Moreover injury , its expitgb3, is upgregulated on cardiomyocytes near the injury site  cells are recruited to the epicardium that are distinct from Wt1+ cells, and after MI these cells are activated and migrate into the subepicardial space  recruitment to the myocardium, causing increased fibrosis, cardiomyopathy and death (2 (PGE2) and its receptor ptger2 were shown to be upregulated after injury. COX enzymes catalyze the rate limiting step in the syntheses of prostanoids such as PGE2, and epicardial cells were observed to have a higher expression of cox2a in their model compared to macrophages and other cardiac cell types , and both transient and stable somatic knockdown of this gene led to defects in the epicardial layer. This suggests tgm2b plays a crucial role in maintaining the integrity of the epicardial sheet during its formation. By creating somatic knock-out animals for genes found in the other subpopulations, the authors observed that sema3fb and cxcl12a had distinct effects on epicardial migration and composition, respectively. Sema3fb was strongly expressed within the bulbous arteriosus (BA), a part of the outflow tract. Sema3fb knockout regulated the number of tbx18-positive cells contributing to SMCs covering the outflow tract. The third population, which was enriched for cxcl12a, was spatially restricted to an area between the BA and atrium. Knockout of this gene revealed that this cell population was involved in homing of leukocytes to the developing heart   within ang heart . Unfortung heart . These dng heart . When long heart . Concordng heart . These dng heart . Additioal layer . Variousng heart , establing heart . The finng heart , indicatng heart . Also, sAnother source of information regarding the epicardial heterogenicity could derive from cardiac cell atlases that have been generated using scRNA seq to identify rare cell populations and interactions within the developing heart. In these studies, the epicardium is often annotated but its potential heterogeneity is often overlooked due to low epicardial cells numbers relative to the total number of cardiac cells, or heterogeneity is ascribed to developmental progression –156. Anoin situ sequencing, it is difficult to draw a concrete conclusion about epicardial composition   . This futiation)  are moretiation) .via paracrine signaling, could result in the identification of mechanisms that aid in cardiac repair. A mouse model of cardiac ischemia/reperfusion (I/R) injury revealed that after 7 days Wt1, Tcf21, and Tbx18 were expressed in distinct as well as in overlapping populations within the subepicardial mesenchyme , d1, d3-5,7, d14, and d28, the evolution of mesenchymal cells was identified. A novel subpopulation was identified dubbed injury response (IR) cells at d1. The IR subpopulation had a high expression of monocyte-macrophage chemoattractants Ccl2, Ccl7, and Csf1 and neutrophil activators Cxcl1 and Cxcl5. Additionally, it also expressed pro-inflammatory and pro-fibrotic factors Il33, Cxcl12, and Tgfb12. These IR cells transitioned to myofibroblasts at d3, and myofibroblasts also displayed heterogeneity. A subpopulation of myofibroblasts was similar to recently identified matrifibrocytes (MFCs) in the mature scar at d14-d28. MFCs were identified as cells that express high levels of ECM genes and support the integrity of the mature scar in injured hearts .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.


To study the effect of a seven-step rehabilitation training program on cardiac function and quality of life in acute myocardial infarction (AMI) patients after percutaneous coronary intervention (PCI).In this study one hundred AMI patients undergoing emergency PCI at The First Hospital of Fangshan District between June 2019 and June 2020 were included. Patients were retrospectively divided into two equal groups based on the type of physiotherapy regiment. The training group included patients who underwent seven-step rehabilitation training while the control group had patients who received routine nursing. Left ventricular ejection fraction (LVEF), self-care capability, hospitalization duration, quality of life, and adverse cardiac event incidence were compared.The number of patients with LVEF values ≥ 50% was significantly higher in the training group after one week of training. Training group patients also showed decreased hospitalization duration and larger improvement in self-care capacity scores. At three months after training, training group patients had overall superior quality of life and lower incidence rates of arrhythmia and angina pectoris.The seven-step rehabilitation training program has a significant effect on improving AMI patient quality of life and cardiac function post-PCI, and is worthy of continued study and promotion. Acute myocardial infarction (AMI) is caused by myocardial necrosis resulting from the interruption—often by coronary atherosclerotic plaques-of continuous myocardial blood supply. Percutaneous coronary intervention (PCI) is widely used clinically as an effective method to quickly restore coronary blood supply. However, while studiesOne hundred patients with AMI that were treated at The First Hospital of Fangshan District with PCI via the radial artery between June 2019 and June 2020 were included in the study. Medical records of the patients were reviewed and the patients were divided into two equal groups (50 patients per group) based on the rehabilitation regiment. A training group received seven-step rehabilitation training, and the control group underwent conventional rehabilitation. The control group contained 39 males and 11 females and averaged 54.2 ± 8.4 years of age. Of these 50 individuals, 28 received one stent, 18 received two, and four received three. Twenty-six of these individuals smoked and 20 consumed alcohols. The training group contained 41 men and 9 women, averaging 56.1 ± 7.5 years of age. Of these 50 individuals, 26 received one stent, 19 received two stents, and five received three stents. Twenty-four of these individuals smoked and 22 consumed alcohols. The study was approved by the Ethics Committee of The First Hospital of Fangshan District.1. Complete medical history2. Patient treatment conformed to the Guidelines for Diagnosis and Treatment of Acute Myocardial Infarction formulated by the Chinese Medical Association: Grade I: patients with cardiac disease but without resulting limitations of physical activity. Grade II: Patients with cardiac disease resulting in slight limitation of physical activity43. Grade I and II cardiac function, based on the New York Heart Association classification4. No chest pain, arrhythmia, or electrocardiogram changes in the eight hours prior to training5. No decompensation indicative of heart failure;6. Myocardial injury marker levels did not further increase after intervention.1. Patients had congenital heart disease2. Chronic obstructive pulmonary disease and other serious lung diseases3. Hepatic and renal insufficiency4. Malignant tumor patients5. Mental illness, consciousness disorder, or inability to cooperateControl group patients received routine nursing consisting of ECG monitoring for myocardial ischemia and arrhythmia over the first 24 hours, routine administration of antiplatelet drugs, blockers, lipid-regulating drugs, and other medications, application of compression bandaging with an elastic bandage, and encouraging patients to manage their diet and hydration habits with the aim of keeping stools unobstructed.The training group received a seven-step rehabilitation training method, following hospital guidelines. Detailed progress report was available for each patient, included in the study. Prior to initiation, patients heart rate was confirmed by ECG to be under 100 beats/minutes, with normal blood pressure and no arrhythmia. Step 1: on the first day of training, patients carried out active and passive exercise of limbs on the bed. Patients were allowed to wash hands, wash face, eat, and use bedpan while in bed. Patients were also allowed to sit up after raising the head of the bed or on their own power for 15-30 min up to twice a day with the help of medical workers. Step 2: on the second day of training, in addition to the previous activities, patients were allowed to wash and wipe themselves by the bed, read for under 15 minutes, and sit up for 15-30 minutes, twice a day. Step 3: on the third day of training, in addition to the previous activities, patients were allowed to walk slowly for 30 meters and try to walk to the toilet. Step 4: on the 4th day of training, in addition to the previous activities, patients walked in place 10-15 times, went to the bathroom by themselves, and tried to wash his body with warm water. Step 5: on the 5th day of training, in addition to the previous activities, patients were allowed to walk 150 meters and tried to take three steps on the training escalator, twice a day, as well as go to the bathroom for cleaning. Step 6: on the 6th day of training, in addition to the previous activities, patients were allowed to walk 150 meters and tried to take five steps on the training escalator, twice a day. Step 7: on the 7th day of training, in addition to the previous activities, patients were allowed to walk 150 meters and tried to take 10 steps on the training escalator, twice a day.The day’s training course was stopped immediately if patients felt palpitations, chest tightness, shortness of breath, dizziness, or other symptoms during training. Patients were not permitted to advance to the next step if any of the following signs were apparent after training: heart rate (as monitored via ECG) ≥ 110 beats/min, systolic blood pressure fluctuations exceeding 20 mmHg from resting values, ST segment elevation) ≥ 0. 2mV or depression) ≥ 0. 1 mV. In these situations, the training plan successfully completed during the prior day would be implemented.All patients in the study were diagnosed with ST Elevation Myocardial Infarction (STEMI). The baseline values of cTnT and cTnI were (1.24 ± 0.13) μ g/L and (25.68±1.56) μ g/L, respectively.Records of baseline and post-training left ventricular ejection fraction (LVEF) measurements obtained using echocardiography were analyzed for each patient. An LVEF under 50% indicated decreased ventricular function.The activity of daily living (ADL) scale was used to assess quality of life before and after training. ADL evaluates a total of 10 activities , with each item scored between 0-10. A maximum score is 100 points, with a score over 60 indicating self-sufficiency. Scores between 41-60 indicate some assistance is required; 21-40 indicates considerable assistance required; and scores under 20 indicate complete dependency. ADL scores of two groups were retrospectively compared.A single follow-up was conducted three months post-discharge. Outpatient nurses conducted one-on-one interviews touching on the areas of environment, physiology, independence, and psychology, using the answers to fill in a standard summary form. Doctors confirmed the presence or absence of angina pectoris, heart failure, arrhythmia, or recurrent coronary stenosis .χ2 is the test comparison result, x±s is the mean and standard deviation of measurement data, t is the comparison result of group test. When p < 0.05, the difference was statistically significant.All data were analyzed using SPSS 19.0 statistical software. The counting data is expressed in the form of rate (n%), No statistical significance in age, gender, number of stents implanted, or history of tobacco/alcohol usage was found between the two groups .There was no difference in baseline LVEF between the two groups prior to training. After training, the number of patients with LVEF values ≥ 50% in the training group was significantly higher than in the control group .- At three months post-training, quality of life for training group patients was better than control group patients  with routine nursing after PCI in AMI patients. It found that after one week of rehabilitation training, the number of patients with LVEF ≥ 50% was significantly higher in patients receiving seven-step rehabilitation training. The early implementation of gradual exercise may enable patients to quickly recover physical fitness and promote cardiovascular regulation under relatively safe conditions. It may also promote the formation of collateral circulation and increase ventricular systolic function, thereby increasing LVEF.This study also found that implementation of the seven-step rehabilitation training program significantly reduced hospital stay durations. In alignment with this, the ability of patients to take care of themselves also improved. It is possible that the incremental elevation of physical activity bears similarities to daily living activities and is conducive to developing self-care capability.These trends persisted at three months following intervention, indicating that early rehabilitation exercise can significantly improve patient self-care capabilities over a prolonged duration, possibly due to promoting the recovery of motor neurons.It is the small sample size of this study from a single center. Although it has certain clinical significance, it needs to further expand the sample validation. In addition, the way of rehabilitation training has more reference, but there is no unified standard. The bias of nursing methods caused by individual factors of patients cannot be avoided. Therefore, the clinical application of its nursing method needs to be further improved.13The seven-step rehabilitation training regimen can significantly improve cardiac function in AMI patients post-PCI, thereby promoting improved quality of life and reduction of adverse cardiac events. However, additional studies are needed to strengthen the findings of this study, given its small sample size.XP and JZ: Conceived and designed the study and is responsible for integrity of the study.LW, HW & WZ: Collected the data and performed the analysis.XP and JZ: Was involved in the writing of the manuscript.All authors have read and approved the final manuscript.


Scrapie is a transmissible spongiform encephalopathy affecting sheep and goats. Due to similarities with mad cow disease (BSE) which is transmissible to humans, scrapie is considered of significance for its theoretical zoonotic potential. A variety of control strategies have been proposed and applied over the years. While herd depopulation, culling of affected susceptible animals, and disinfection measures are poorly successful, selective breeding schemes focused on amino acid substitutions (polymorphisms) in prion protein (PrP) would be the best way for eradication. In sheep, the association between PrP gene polymorphism argininine on codon 171 is highly effective for scrapie resistance breeding. Although in goats this polymorphism is absent, three other candidate PrP amino acid polymorphisms have been recognized for resistance breeding. For Greece with the largest goat population in Europe, the frequencies of these polymorphisms has yet to be sufficiently documented. This study performed a large-scale, cross-sectional survey for a large part of the country, regarding PrP polymorphic codons 110, 146, 211, and 222, and estimated effects of these polymorphisms on (i) the likelihood to be infected by scrapie, and (ii) scrapie status at the herd level, revealing possible candidate PrP amino acid polymorphisms to be used in breeding for scrapie-resistance programs.n = 525) from scrapie-affected (n = 282) and non-affected (n = 243) animals within the national surveillance program, and (ii) individual blood samples (n = 1708) from affected (n = 241) and non-affected (n = 1467) herds, in a large part of mainland Greece and its islands, were collected and assayed. A dedicated Taqman method was used to test for amino acid polymorphisms 110T/P, 146N/S/D, 211R/Q, and 222Q/K. Highly prevalent genotypes were 110TT, 146NN, 211RR, and 222QQ. The frequencies of polymorphisms in blood and negative brain samples for codons 110P, 211Q, and 222K were 4.0%, 3.0%, and 1.9%, respectively, while 146D (0.7%) was present only on Karpathos island. Codon 110P was exclusively found in scrapie-negative brains, and homozygous 110P/P in two scrapie-negative goats. It is concluded that breeding programs in Karpathos could focus on codon 146D, while in other regions carriers of the 110P and 222K allele should be sought. Case-control and challenge studies are now necessary to elucidate the most efficient breeding strategies.Scrapie is considered an endemic disease in both sheep and goats in Greece. However, contrary to sheep, in goats more than one prion protein (PrP) polymorphism has been recognized as a candidate for resistance breeding against the disease. For an impression, candidates which are circulating, (i) brain samples ( Dairy goat farming in the EU ranks among the dynamic and promising livestock sectors, growing fast to satisfy the increasing demand for goat milk and dairy products . Its mulSc) of the normal cellular (PrPC) prion protein (PrP), which is a cell–surface glycoprotein. The transmission and the incubation period of the disease depend on the exposure to the infectious agent, the scrapie strain, and the genetic background of the host [Sc biochemical properties. Control strategies which have been utilized over the years for small ruminants include: (i) Complete herd depopulation, (ii) depopulation of genetically related animals , (iii) depopulation of animals at risk, (iv) culling of affected animals, (v) cleaning and disinfection of premises and equipment, (vi) sufficient period until animals will be restocked in the premises, and (vii) selective breeding schemes based on the PrP gene polymorphisms  [Scrapie is among these diseases and in Greece has been diagnosed since 1987 in sheep and 1997 in goats . It is ascrapie) ,11,12.6 milking goats (ca. 30% of the EU milking goat herd) producing about 356,000 tons of milk , mainly used to produce Feta cheese [Greece has the largest national goat flock in Europe with more than 2.6 × 10PRNP gene that is coding for the prion protein named PrP  [C conversion into the pathogenic isoform PrPSc [In sheep, selective breeding schemes have been exploited, to eliminate the prevalence of scrapie in the EU, based on the selection of a resistance-related polymorphism in the 99/2001) . Severalrm PrPSc ,20,21,22rm PrPSc  and exemrm PrPSc . PRNP gene polymorphisms have been found, however, their role regarding the incubation period and the susceptibility to classical scrapie has not been adequately documented. Atypical scrapie has also been detected in goats in many countries including Switzerland, France, Spain, and Italy. Cases of atypical scrapie are distinguished by a 5-band profile on Western blot with a prominent lower band at approximately 11 to 12 kDa, whereas, classical scrapie has the nonglycosylated band at approximately 19 to 21 kDa. In addition, the PrPSc of atypical scrapie is relatively sensitive to proteinases [Similarly to sheep, goat PrP is very polymorphic, but the critical arginine at codon 171 used in sheep for scrapie-resistance breeding does not occur in goats ,27,28,29teinases ,31. In tteinases ,38,39,40teinases ,41. HoweEstimating the frequency of candidate resistant alleles and quantifying their effect is a preliminary and crucial step in determining the feasibility, impact, and cost-effectiveness of the proposed selective breeding programs . The objPRNP gene locus. Brain sampling took place according to the European Parliament and Council Regulation EC no. 999/2001 [A total of 282 and 243 brain samples from scrapie-affected (64 herds) and non-affected (11 herds) animals were collected, respectively, within the national TSEs surveillance program between years 2008–2014 and analyzed to detect polymorphisms at the 999/2001  from then = 12 herds, 241 samples) and non-affected  herds. Enrolled herds were in the prefectures of Macedonia and Thessaly, Dodecanese and Ionian Islands, and Thrace , which were sequenced by the Sanger method in an ABI 3500 Genetic Analyzer (Applied Biosystems) performing direct sequencing of the full coding region. From all samples (brain and blood), genomic DNA was extracted using the PureLink Genomic DNA Mini Kit (Life Sciences) according to the manufacturer’s instructions. The d 222Q/K , oligonuo is the observed and e is the expected number of each genotype assuming the Hardy-Weinberg equilibrium. The effects of the 146N/S/D, 211R/Q, and 222Q/K polymorphisms (i) on the herd’s scrapie-status , and (ii) on the animal’s scrapie-status  were assessed using the following binary regression model:Allelic and genotypic frequencies were calculated with counting. Allelic frequencies were used to calculate the expected numbers of genotypes under the assumed Hardy-Weinberg equilibrium. The observed and expected numbers of genotypes were compared using the chi-square test as described below:Blood and brain samples were either routinely collected for the interest of the national scrapie eradication program or as a part of the non-experimental standard clinical veterinary practice. In any case, this study followed the European Directive 86/609/EEC and its national implementation by the Greece Presidential Decree no. 160/1991 .Frequencies of genotypes and alleles at codons 110, 146, 211, and 222 for (i) brain samples from scrapie-positive and scrapie-negative goats, and (ii) blood samples from scrapie-positive and scrapie-negative herds are summarized in p = 0.065, 95% CI, 0.9–50.0). Moreover, the 211RQ genotype was ca. 5.6 times more likely to be observed in scrapie-positive herds when compared to goats with the 211RR genotype , whereas, frequencies of polymorphisms at codons 146 and 222 were not found to significantly differ between scrapie-positive and scrapie-negative herds.Attempts to propose breeding for scrapie resistance programs in Greece have been made by Vouraki et al.  and KanaAt codon 110, genotype 110TP was found in both scrapie-negative and scrapie-positive farms and in scrapie-negative animals but in no case in scrapie-positive animals. This is a strong indication of the protective effect of genotype 110TP, though it was not possible to be statistically proven, as the effect of codon 110 on scrapie status was not estimated by the model. In any case, further testing in experimental challenge studies is necessary to confirm this assumption. If genotype 110TP is proved to exhibit a protective action against scrapie its inclusion in EU legislation needs to be supported by EFSA. Moreover, it is the first time that the homozygous genotype 110PP is observed in goats in Greece . p = 0.065). These findings are in accordance with the available literature, where a lower susceptibility of both 146ND and 146NS genotypes and a higher genotype for 146NN to natural and experimental scrapie infection have been observed in field studies [Due to the absence or very low frequencies of alleles D146 and S146 in Europe, except from Cyprus, no results from field studies regarding resistance of genotypes at codon 146 to European scrapie strains have been available up to now . This ha studies ,28,32,35 studies  studied p < 0.001). It could be assumed that the 211RQ genotype may be associated with susceptibility to the disease at the herd level. However, this assumption is not supported by the model used to compare genotypes at the animal level, where no differences were observed. Additionally, it is not supported by the available literature. For example, Barillet et al. [In the case of codon 211, it was found that goats with the 211RQ genotype were ca. 5.6 times more likely to originate from a scrapie-positive herd when compared to goats with the 211RR genotype , which is indicative for a rather high resistance of 222K to PrPC>PrPSc conversion [PRNP gene sequences that the 222K allele contains mutations associated with susceptibility. Moreover, the fact that (i) all samples derived from mixed flocks with a high prevalence of sheep scrapie, indicating a strong infection pressure, (ii) the existence of the disease for more than 2 years, and (iii) the potential occurrence of different scrapie strains, need to be considered as significant risk factors and possible confounders. Another study design using farm rather than animal as a statistical unit might address this issue. Moreover, an accurate case-control study in important goat dairy production regions is still required to confirm the quality of the 222K codon.Field and surveillance data regarding scrapie occurrence and the evaluation of candidate alleles that confer resistance/susceptibility to the disease, have indicated that the 222K allele is associated with strong resistance against it across Europe ,48,49,50nversion . In our In our study, no homozygous 222K goats were found to estimate the potential protective effect of the genotype 222K. In general, the 222K allele occurs in many dairy goat breeds, but at rather low frequencies in most of the European countries, e.g., only 0.6–0.7% in different breeds . This isBased on our findings and the results from relevant studies in Greece, it could be concluded that breeding programs could be based on the codon 146 for Karpathos island and 222 for the mainland. Codon 110 needs to be also considered as a promising one, however more evidence regarding its performance need to be acquired. Furthermore, codon 146D could also be included in these inventory studies . Therefo


Although physical activity maintenance is important for OA management, it is not clear whether people with OA are more inactive or not. One possible reason is no simple monitoring tool to assess physical activity at the clinic. The aim of this study was to determine the reliability and validity of the Baecke Physical Activity Questionnaire (BQ) in adult women with hip disorders.Sixty-four patients with unilateral or bilateral hip disorders were recruited from an outpatients clinic at a university hospital in Japan. BQ includes a total of 16 questions classified into three domains: work, sports, and non-sports leisure activity. For test-retest reliability, one BQ was administrated face-to-face, and a second was mailed to participants two weeks later. Test-retest reliability of BQ was assessed using intra-class correlation (ICC) and Bland and Altman method. To determine criterion validity, the correlation between BQ measurements and pedometer-measured step counts was assessed. Correlations between BQ measurements and step counts were assessed using Spearman rank correlation coefficient (rho).Analyses were restricted to the 61 patients (53.3 ± 11.3 years old) who wore the pedometer continuously for 5 days or more. Twenty eight patients had unilateral hip osteoarthritis, 17 patients had unilateral total hip arthroplasty, and 16 patients had hip osteoarthritis and total hip arthroplasty. The mean step count was 6,309 ± 2,392 steps/day. In analysis for reliability, the value of ICC was 0.84 for work, 0.83 for sports, 0.78 for non-sports leisure activity, and 0.87 total. Bland and Altman analysis showed the step count and BQ total did not differ significantly from 0 with most falling between 0 ± 1.96 SD. In analysis for validity, there was a significant but low to moderate correlation between step counts and 3 BQ subscales  and a higher correlation between step counts and total score .BQ is a useful monitoring tool for assessing multiple domains of physical activity with acceptable reliability and validity in adult women with hip disorders. Osteoarthritis (OA) is the most common form of arthritis and is a leading cause of mobility-related disability in older adults . The curIt is well established that physical inactivity is a risk factor for cardiovascular disease, diabetes, osteoporosis, and some cancers . TherefoPhysical activity, however, is a complex and multidimensional exposure variable, making population-based measurement difficult . AlthougAmong many questionnaires, the Baecke Questionnaire (BQ)  is one oSixty four consecutive participants were recruited when visiting an outpatient's clinic at a university hospital. The inclusion criteria were adult women with OA in one or both hips who had or had not undergone past salvage surgery such as femoral osteotomy, and those who had undergone THA for pre-existing OA. All patients were community-dwelling and receiving no regular nursing care. Patients were limited to women because of their great predominance in the clinic. Radiographic evidence of joint space narrowing and associated bony changes confirmed the presence of OA. All surgeries were performed at least 6 months before study entry so that rapid functional changes, which usually occur in the six month postoperative period, could be minimized during the study. The exclusion criteria were symptoms in other weight-bearing joints, and/or chronic diseases which could possibly affect mobility. As all data were collected as part of medical care, no ethical approval was requested. All patients were given experimental information in compliance with Helsinki Declaration and only those who gave consent participated in the study.The Harris Hip Score (HHS) was used to evaluate overall function of the hip, due to its wide use as an outcome measurement after hip surgery. HHS has 4 domains: function, pain, deformity, and range of motion, allowing a total score ranging from 0 to 100 points (low-high), which are categorized as follows: 0 to 70 points, poor; 70 to 80 points, fair; 80 to 90 points, good; 90 to 100 points, excellent .BQ includes a total of 16 questions classified into three domains: work, sports, and non-sports leisure activity. Each domain has several questions scored on a five-point Likert scale, ranging from never to always or very often. Scoring of the questionnaire in our study followed the original system; work was the mean score among eight occupational questions, sports was the mean score among four sports-related questions, and non-sports leisure was the mean score among four habitual physical activities during leisure time. Each domain could receive a score from one to five points, thus allowing a total score from three (minimum) to fifteen (maximum). For the two most frequently reported sports activities, specific questions regarding the number of months per year and hours per week of participation were addressed. The original BQ was translated into Japanese using the forward and backward translation procedure. Two professional translators independently translated the original scale once. Synthesis of the translations was conducted by all members of the study team. For test-retest reliability, the initial BQ was administered during face-to-face consultations, and a second was mailed to participants two week later, and they were asked to return it by mail within a few days. Three subscales "work", "sports", "non-sports leisure" were scored and totaled.Participants were asked to wear a digital pedometer with a uniaxial acceleration sensor  at the waist, during waking hours except during bathing or swimming and to do ordinary daily activities without reading the step count. A pedometer estimates the number of steps taken based on acceleration signals. Although the reproducibility and validity of the pedometer in counting walking steps has been established in healthy people , we confTest-retest reliability of BQ was assessed using intra-class correlation (ICC)  and BlanOf 64 patients, 46 were considered to have worn the pedometer continuously from morning until night for 7 days, 9 for 6 days, 6 for 5 days, 2 for 4 days, and 1 for 3 days during the measurement period. Analysis were restricted to the 61 patients data who wore the pedometer for 5 days or more,2. Twenty eight patients (46%) had unilateral hip osteoarthritis, 17 patients (28%) had unilateral total hip arthroplasty, and 16 patients (26%) had hip osteoarthritis and total hip arthroplasty. The mean step count was 6309 (2392) steps/day. The mean HHS score was 79.3 (12.7). Of the 61 patients, 16 (26%) were categorized as poor HHS, 15 (27%) were fair, 16 (26%) were good, and 14 (23%) were excellent. The mean subscales of initial BQ were 2.8 (SD 0.6) for work, 2.1 (SD 0.6) for sports, 2.6 (SD 0.6) for non-sports leisure, and 7.5 (SD 1.4) for total. From initial BQ, 23 patients (38%) reported engagement in sports. Eighteen of them (78%) were engaged in aqua exercise, such as swimming or aqua walking. The other sports were muscle strengthening exercises and using an exercise bike.Table Of the 61 participants who completed the first BQ, 52 (85%) returned the second BQ. The nine patients who did not return it had no specific characteristics of age, HHS, or step counts. Table Table This study first evaluated the reliability and validity of physical activity questionnaire in adult women with hip osteoarthritis, total hip arthroplasty, or both. BQ showed an acceptable reliability and validity and may be a useful tool for assessing physical activity in patients with hip disorders.Three subscales and total score of BQ showed high correlation coefficients between test and re-test, which is consistent with the literature ,24. In tWe used step count assessment by pedometer as a reference tool. Walking is the most frequently reported leisure-time physical activity in patients with arthritis . AdditioThe magnitude of BQ correlations varies substantially among different study samples and instruments measuring physical activity in the literature. For example, a total score of BQ was highly correlated with a measure using double labeled water (Pearson's r = .69), the gold standard in the assessment of physical activity, in men aged 40 years . HoweverThere are two limitations of this study. First, the pedometer could not be worn during aqua exercise. Eighteen patients reported engagement in aqua exercise, such as swimming or aqua walking. Therefore, sport index may have had the lowest correlation coefficient, among BQ measurements. Second, the pedometer and BQ were different in terms of reference periods. The pedometer was used for only 5 to 7 days, whereas BQ referred to activities of no specified time component.Previous studies have shown that mean BQ measures are 2.9 for work, 2.4 for sports, and 3.1 for leisure in women aged 20–32 years  and 2.7,In conclusion, BQ is a useful tool for assessing multiple domains of physical activity with acceptable reliability and validity in adult women who have hip disorders.The author(s) declare that they have no competing interests.RO carried out the validation and reliability study, participated in the sequence alignment and drafted the manuscript. SH, MK, YT conceived of the study, and participated in its design and coordination and helped to draft the manuscript. MY, TN, participated in acquisition of data at the outpatient's clinic and helped to interpret these data. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here: