Objective To determine whether local gradient fluid shear stress(FSS)causes a specific distribution of intracellular calcium ion concentration,which ultimately determines the direction of cell migration.Methods Numerical simulations were performed using COMSOL software.The method of staining intracellular calcium ion for RAW264.7 osteoclast precursors was established.After applying gradient FSS on the cells,the distribution and dynamic changes of intracellular calcium ion concentration and cell migration parameters were analyzed.Results Osteoclast precursors tended to migrate towards regions with lower FSS,and oscillatory flow regulated the distribution of intracellular calcium ions along the direction of cell migration.After blocking phospholipase C(PLC),mechanosensitive cation-selective channels(MSCC),endoplasmic reticulum(ER),and removing extracellular calcium,the migration speed of cells towards the low FSS direction was significantly reduced,but the migration speed along the liquid flow direction was significantly enhanced.Meanwhile,the calcium ion distribution along the liquid flow direction was significantly increased.Conclusions Osteoclast precursors can sense the FSS gradient,resulting in a specific distribution of intracellular calcium ions along the direction of migration.This ultimately leads to the migration of osteoclast precursors towards regions with lower FSS.This study provides important basic data for ultimately elucidating the cellular and molecular mechanisms of bone tissue remodeling under dynamic external forces.

Objective:To explore the prevalence and independent associated factors of vascular calcification (VC) in non-dialysis chronic kidney disease (CKD) patients of stage 3-5.Methods:It was a single-center cross-sectional observational study. Non-dialysis stage 3-5 CKD patients ≥18 years old who were admitted to the Department of Nephrology, the First Affiliated Hospital of Sun Yat-sen University from May 1, 2022 to December 31, 2022 with VC evaluation were enrolled. The patients' general information, laboratory examination and imaging data were collected. Coronary artery calcification (CAC), thoracic aorta calcification (TAC), abdominal aorta calcification (AAC), carotid artery calcification and aortic valve calcification (AVC) were evaluated by cardiac-gated electron-beam CT (EBCT) scans, lateral lumbar x-ray, cervical macrovascular ultrasound and echocardiography, respectively. The differences in clinical data and the prevalence of VC at different sites of patients with different CKD stages were compared, and the prevalence of VC at different sites of patients in different age groups [youth group (18-44 years old), middle-aged group (45-64 years old) and elderly group (≥65 years old)] and patients with or without diabetes were compared. Multivariate logistic regression analysis was used to analyse the independent associated factors of VC for different areas.Results:A total of 206 patients aged (51±14) years were included, including 129 (62.6%) males. There were 44 patients with CKD stage 3 (21.4%), 51 patients with CKD stage 4 (24.8%), and 111 patients with CKD stage 5 (53.9%). CKD was caused by chronic glomerulonephritis [104 cases (50.5%)], diabetic kidney damage [35 cases (17.0%)], hypertensive kidney damage [29 cases (14.1%)] and others [38 cases (18.4%)]. Among 206 patients, 131 (63.6%) exhibited cardiovascular calcification, and the prevalence of CAC, TAC, AAC, carotid artery calcification, and AVC was 37.9%, 43.7%, 37.9%, 35.9% and 9.7%, respectively. The overall prevalence of VC in young, middle-aged and elderly patients was 24.6%, 73.6% and 97.4%, respectively. With the increase of age, the prevalence of VC in each site gradually increased, and the increasing trend was statistically significant (all P<0.001). The overall prevalence of VC in CKD patients with diabetes was 92.5% (62/67), and the prevalence of VC at each site in the patients with diabetes was significantly higher than that in the patients without diabetes (all P<0.001). Multivariate logistic regression analysis revealed that age (every 10 years increase, OR=2.51, 95% CI 1.77-3.56, P<0.001), hypertension ( OR=5.88, 95% CI 1.57-22.10, P=0.009), and diabetes ( OR=4.66, 95% CI 2.10-10.35, P<0.001) were independently correlated with CAC; Age (every 10 years increase, OR=6.43, 95% CI 3.64-11.36, P<0.001) and hypertension ( OR=6.09, 95% CI 1.33-27.84, P=0.020) were independently correlated with TAC; Female ( OR=0.23, 95% CI 0.07-0.72, P=0.011), age (every 10 years increase, OR=3.90, 95% CI 2.42-6.29, P<0.001), diabetes ( OR=5.37, 95% CI 2.19-13.19, P<0.001) and serum magnesium ( OR=0.01,95% CI 0-0.35, P=0.014) were independently correlated with AAC. Moreover, age and diabetes were independently correlated with carotid artery calcification, AVC and overall VC Conclusions:The prevalence of VC in non-dialysis CKD patients of stage 3-5 is 63.59%, of which CAC reaches 37.9%, TAC is the most common one (43.7%), while AVC is the least one (9.7%). Age and diabetes are the independent associated factors for VC of all sites except TAC, while hypertension is an independent associated factor for both CAC and TAC.

Objective By comparing the fatigue strength of type A and type B locking compression plates (LCP) in distal femoral plate, a theoretical evaluation method was provided for type selection of bone plate when testing its bending strength and fatigue performance. Methods Through bending strength performance test and fatigue performance test on bone plates with different types, combined with ANSYS Workbench, the finite element analysis on total deformation, von Mises stress and fatigue service life of bone plates were conducted. Results The fatigue strength of type A plate was 30.7% higher than that of type B plate, the stress of type A plate was lower than that of type B plate, and the minimum fatigue service life of type A plate was 17% higher than that of type B plate. Conclusions The fatigue performance of type A plate is better than that of type B plate, so the failure possibility of type A plate was lower than that of type B plate.The results provide references for assisting selection of different bone plates when testing the performance of two newly developed bone plates.

Objective To express the monkeypox virus (MPXV) A23R protein in Escherichia coli and purify by Ni-NTA affinity column, and to prepare mouse antiserum against MPXV A23R. Methods The recombinant plasmid pET-28a-MPXV-A23R was constructed and transformed into Escherichia coli BL21 to induce the expression of A23R protein. After optimizing the conditions of expression, A23R protein was highly expressed. Recombinant A23R protein was purified by Ni-NTA affinity column and identified by Western blot analysis. The purified protein was used to immunize mice for preparing the A23R polyclonal antibody, and the antibody titer was detected by ELISA. Results The expression of A23R recombinant protein reached the peak under the induced conditions of 0.6 mmol/L isopropyl-β-D-thiogalactoside (IPTG), 37 DegreesCelsius and 20 hours. The purity of the protein was about 96.07% and was identified by Western blot analysis. The mice were immunized with recombinant protein, and the titer of antibody reached 1:102 400 at the 6th week after immunization. Conclusion MPXV A23R is expressed highly and purified with a high purity and its antiserum from mouse is obtained with a high titre.
Animals ; Mice ; Monkeypox virus ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Blotting, Western ; Recombinant Proteins ; Escherichia coli/genetics*

Objective:To study the effect of excessive torsion of the umbilical cord on fetal or neonatal outcomes.Methods:The observation group was selected from the puerperae who delivered in Beijing Obstetrics and Gynecology Hospital from July 2016 to June 2020 with excessive torsion of the umbilical cord. In the same period, the puerperae without excessive torsion of the umbilical cord were selected as the control group with a ratio of 1∶1. The general condition, mode of delivery, perinatal outcomes, and the effect of different umbilical coiling index (UCI) [twisted umbilical cord weeks/umbilical cord length (cm)] on fetal and neonatal outcomes were retrospectively analyzed between two groups.Results:Compared with the control group, the observation group (1 780 cases) had smaller neonatal gestational age [(37.9±3.2) weeks vs. (38.4±2.9) weeks], birth weight [(3 007±726) g vs. (3 354±616) g] and length [(48.5±4.3) cm vs. (49.6±4.1) cm], but higher incidence of fetal distress [34.9% (622/1 780) vs. 12.9% (230/1 780)], neonatal asphyxia [1.5% (26/1 780) vs. 0.7% (13/1 780)], and cord blood pH<7.20 [4.2% (75/1 780) vs. 2.8% (49/1 780)], the difference was statistically significant ( P<0.05). The UCI≥0.73 group had lower neonatal gestational age, birth weight and length, but higher incidence of cord blood pH<7.20 and neonatal asphyxia than the UCI<0.73 group, with statistically significant differences ( P<0.05). Conclusions:Excessive torsion of the umbilical cord increases the incidence of fetal hypoxia and neonatal asphyxia and has a significant effect on neonatal gestational age, birth weight and length. The higher the UCI, the greater the impact on fetus and neonate.

Objective:To investigate the biomechanical characteristics and clinical outcomes of arthroscopic superior capsular reconstruction for massive irreparable rotator cuff tears (MIRCT) using the long head of biceps tendon (LHBT) with tenotomizing its distally or not (the "Chinese way" ).Methods:Eight fresh-frozen cadaveric shoulders were used to create a MIRCT model by detaching the footprints of the supraspinatus and infraspinatus tendons on the greater tuberosity. LHBT autograft was transferred and securely fixed onto the footprint of supraspinatus tendon for superior capsular reconstruction. Further, all cadaveric specimens were assigned to the tenotomy group or reservation group (4 cadaveric specimens in each group) according to whether the distal part of LHBT was tentomized or not. Biomechanical tests were conducted to observe the stiffness, ultimate load of fixed LHBT and to measure the length between LHBT tear site and its insertion on the superior labrum. A total of 41 patients with MIRCT who underwent arthroscopic superior capsular reconstruction using LHBT autograft between July 2016 and December 2018 were enrolled in the study. There were 17 males and 24 females, aged from 46 to 76 years (62.6±7.3 years). All patients were assigned to the tenotomy group (23 cases) or reservation group (18 cases) according to whether the distal part of LHBT was tentomized or not. The visual analogue scale (VAS), University of California, Los Angeles (UCLA) score, American Shoulder and Elbow Surgeons (ASES) score, Constant-Murley score and Fudan University Shoulder Score (FUSS) were used to evaluate the clinical outcomes. The range of motion (ROM) of shoulder was recorded before and after operation. Magnetic resonance imaging was used to assess the structural integrity of reconstructed tissue at 12 months after operation (refers to the failure of the transposed LHBT, which may be accompanied by a retear of partial repaired supraspinatus tendon).Results:Biomechanical research showed that the stiffness, ultimate load of fixed LHBT and the length between LHBT tear site and its insertion on the superior labrum in the reservation group (54.0±6.6 N/mm, 141.8±15.9 N, 93.3±12.4 mm, respectively) were significantly higher than those in the tenotomy group (25.7±4.2 N/mm, 80.8±8.0 N, 47.4±2.0 mm, respectively) ( P<0.05). All patients were followed up for 12-18 months (14.5±1.8 months) without significant complications and adverse reactions. No matter the distal part of LHBT was tentomized or not, the ROM and clinical scores (VAS score, UCLA score, Constant-Murley score, ASES score and FUSS) of patients improved significantly at 1 year follow-up than that before operation ( P<0.05). However, there were no significant differences between the reservation group and tenotomy group in terms of postoperative ROM [flexion, abduction, external rotation at side, internal rotation (vertebral level) were 144.3°±15.5° vs. 148.0°±10.3°, 145.1°±14.1° vs. 142.3°±11.2°, 67.3°±14.4° vs. 62.7°±11.7°, 8.3±2.1 vs. 7.8±2.5, respectively], VAS scores (2.3±1.6 vs.1.5±1.2), functional scores (Constant-Murley score, UCLA score, ASES score and FUSS were 88.2±11.4 vs. 85.6±9.6, 29.3±2.8 vs. 31.4±3.5, 86.8±11.8 vs. 82.6±9.2, 92.1±10.1 vs. 88.3±8.2, respectively) and structural failures (35.2% vs. 30.0%, P>0.05). Conclusion:Arthroscopic superior capsular reconstruction using LHBT with reserving its distal part could achieve higher mechanics strength. However, the short-term follow-up showed that tenotomizing the distal part of LHBT exerted no obvious influence on postoperative function and structural integrity.

Objective: To analyse the outcomes and the prognostic factors of patients with sinonasal malignancies following endoscopic endonasal approach, and to compare the pre- and post-operative quality of life. Methods: A retrospective single-center review of 79 patients who underwent endoscopic endonasal approach for sinonasal malignancies in Beijing Tongren Hospital from October 2004 to March 2017 was performed, including 51 males and 28 females, with a median age of 48 years. Data of demography, imaging (including nasal CT and MRI before operation), histopathology and treatment strategy were collected. Recurrence and distant metastasis were diagnosed according to endoscopic examination, MRI and general check-up after surgery. Pre- and post-operative quality of life scores were obtained by sinonasal outcome test-22, visual analog scale and anterior skull base surgery questionnaire. SPSS 22 software was used for statistical analysis. Results: The study consisted of 13 pathological types with sinonasal T1-T4 stage tumors, including cervical lymph nodes and/or distant metastasis. All patients underwent endoscopic endonasal approach surgery. After 43 months of median follow-up time, the overall, disease-free, and recurrence-free survival rates at 1, 3, 5 and 10 years was 97.4%, 92.5%, 92.5% and 83.7%; 83.2%, 68.3%, 56.8% and 33.6%; 84.5%, 66.6%, 58.0% and 34.4%, respectively. Postoperative recurrence was an independent risk factor affecting the overall survival rate (HR=8.852, P=0.044), and preoperative recurrence (secondary surgery) was an independent risk factor affecting the disease-free and recurrence-free survival rate (HR value was 2.237 and 2.095 respectively, P value was 0.029 and 0.047 respectively). After surgery, the olfaction and nasal scab got worse, while the nasal obstruction and breathing were improved. Conclusions Endoscopic endonasal approach for sinonasal malignancies can achieve satisfactory outcomes, and has obvious advantages in improving the quality of life. Postoperative recurrence and preoperative recurrence are the prognostic factors.

Objective:To study the significance of induction chemotherapy and subsequent comprehensive therapy for overall survival rate (OS) and larynx dysfunction-free survival rate (LDFS) in patients with advanced hypopharyngeal carcinoma.Methods:Patients who met the inclusion criteria with the diagnoses of advanced hypopharyngeal carcinoma between 2011 and 2017 received 2 or 3 cycles of TPF regimen induction chemotherapy. Patients who attained complete response (CR) received radical chemotherapy. Patients who attained partial response (PR) and the reduction of tumor volume was more than 70% were defined as large PR and received concurrent chemoradiotherapy. When the tumor volume reduction of PR patients was less than 70%, they were defined as small PR. (CR+large PR) group was defined as effective group. Patients who did not reach CR and large PR were defined as uneffective group and underwent radical surgery and received adjuvant radiotherapy as appropriate after the surgery. The end points of the study were OS, progression-free survival (PFS) and LDFS. Chi-square (χ 2) test was used for correlation analysis. Survival analysis was performed by the Kaplan-Meier method with a Log-rank test. Cox proportional hazards model was used for univariate and multivariate survival analysis. Results:A total of 260 patients were enrolled in the study. The follow-up period ranged from 5 to 83 months, with an average of 24.7 months. The 3-year and 5-year OS rate was 46.0% and 32.6%, respectively. The 3-year and 5-year PFS rate was 41.0% and 26.6%, respectively. The 3-year and 5-year LDFS rate was 37.9% and 24.8%, respectively. Poor outcome of induction chemotherapy, advanced N stage, strong positive Ki-67 immunohistochemistry (all P<0.001) were negative prognostic factors. The advanced clinical stage was positively related to the poor outcome of induction chemotherapy ( P=0.015). There was no significant difference in OS and PFS between the large PR group and the small PR group (all P>0.005). Conclusion:TPF regimen induction chemotherapy and subsequent comprehensive therapy for patients with advanced hypopharyngeal carcinoma may improve the quality of life of patients, with high OS rate and LDFS rate.

OBJECTIVE： To investigate the protective effect and its mechanism of thymoquinone （TQ） on myocardial fibrosis （MF） induced by lipopolysaccharide in rats. METHODS： Totally 40 male Wistar rats were randomly divided into control group， model group， TQ low-dose and high-dose groups [5， 10 mg/（kg?d）]， with 10 rats in each group. Except that control group was given constant volume of normal saline intraperitoneally， other groups were given LPS intraperitoneally to induce MF model， once a day， for consecutive 3 weeks. Since the first day of modeling， administration group was given relevant medicine intraperitoneally， once a day， for consecutive 3 weeks. After last medication， ELISA method was used to detect the contents of serum inflammation factors （IL-1β， IL-6， TNF-α） in rats. Cardiac mass parameters （HW/BM， LVW/BW） were weighed and calculated. HE staining was used to observe the histopathological changes of myocardial tissue. The contents or activities of oxidative stress indicators （MDA， SOD） and myocardial collagen indexes （HYP， Col-Ⅰ， Col-Ⅲ） were detected by chemical analysis， xanthine oxidase method or ELISA. mRNA expression of regulation genes （Col-Ⅰ， Col-Ⅲ， MMP-3， MMP-9， TGF-β1 and Smad3） related to myocardial fibrosis were determined by real-time fluorescence quantitative PCR. RESULTS： Compared with control group， there were swollen myocardial cells， disordered nuclei of different sizes and visible fiber hyperplasia in model group； the levels of serum inflammatory factors and LVW/BW， cardiac contents of MDA， HYP， Col-Ⅰand Col-Ⅲ， mRNA expression of Col-Ⅲ， TGF-β1 and Smad3 in model group were increased significantly （P＜0.05）， while SOD activity was decreased significantly （P＜0.05）. Compared with model group， the degree of myocardial fibrosis was improved in TQ groups to different extents； serum content of IL-1β and the contents of MDA， HYP and Col-Ⅰ in cardiac tissue in TQ low-dose group， serum contents of IL-1β， IL-6 and TNF-α， LVW/BW and the contents of MDA， HYP， Col-Ⅰ and Col-Ⅲ in cardiac tissue of TQ high-dose group as well as mRNA expression of Col-Ⅰ， Col -Ⅲ， TGF-β1 in TQ groups were decreased significantly （P＜0.05）. The activities of SOD in cardiac tissue were increased significantly in TQ groups （P＜0.05）. There was no statistical significance in other indexes among groups （P＞0.05）. CONCLUSIONS： TQ can protect against MF model rats to certain extent， the mechanism of which may be associated with the inhibition of inflammation reaction and oxidant stress reaction， and down-regulation of mRNA expression of Col-Ⅰ， Col-Ⅲ and TGF-β1.

Objective To conclude the experience of restoration of fat compartments volume intreatment of facial volume lost aging.Methods From May 2015 to May 2018,126 patients with facialvolume lost aging,aged from 34 to 52 years (mean age 42.6 years),were corrected by restoring fatcompartments volume using different sized autologous fat granule grafting technique.Patients' satis-faction was rated using Visual Analogue Scale (VAS) ranging from 1 (most unsatisfactory) to 10(most satisfactory).Results 126 patients with volume lost aging face were followed up from 3months to 2.5 years,the sculpted appearance became more fullness,the laxity,wrinkles and textureof skin were improved obviously,the patients' VAS score was (8.2±1.4).Conclusions Restorationof fat compartments by using different sized autologous fat granule grafting can augment the facicalfullness,reshape the facial contour and improve the aging skin.Therefore it proves to be a safe,multi-effecive and satisfying technique in the treatment of facial volume lost aging.