[Summary] The activities of 17α-hydroxylase and 17,20 lyase of cytochrome P450c17 were evaluated by ELISA in NCI-H295R cells after treatment with palmitate and oleate. The results showed that 0. 75 mmol/L plamitate did not influence the activity of 17α-hydroxylase, but increased the activity of 17, 20 lyase by 74. 3% ( P<0. 05). Oleate at the same concentration did not change the activity of 17,20 lyase. There were no significant changes in the protein expressions of P450c17, P450 oxidoreductase, and cytochrome b5 after treatment with palmitate and oleate. However,reactive oxygen species in cells were elevated by palmitate. The results suggest that exposure to palmitate may increase androgen production by inducing 17, 20 lyase activity of P450c17 in NCI-H295R cells, which is related with oxidative stress-mediated post-translational regulation of the enzyme.

To establish an LC-MS/MS method for simultaneous determination of peimine and peiminine in rat plasma after oral and intravenous administration of Fritillaria thunbergii Miq. extract, the pharmacokinetic parameters were calculated as well. Peimine, peiminine and internal standard carbamazepine were extracted from plasma with liquid-liquid extraction by ethyl acetate, then separated on a Luna C18 column by using acetonitrile-water containing 10 mmol x L(-1) ammonium formate (35:65), as mobile phase. The electrospray ionization (ESI) source was applied and operated in positive ion mode. Peimine was detected at m/z 432.4 --> 414.4, peiminine at m/z 430.4 --> 412.4 and carbamazepine (IS) at 237.1 --> 194.2. The linear calibration curves were obtained at the concentration range of 0.8-800 ng x mL(-1) for peimine and peiminine. The extraction recoveries were 94.1%-105.3% and 85.8%-98.6%, respectively. The precisions, accuracy and stability of the analytes meet the requirements. The method was shown to be effective, convenient, and suitable for simultaneous pharmacokinetic study of peimine and peiminine in rat.

Objective To investigate the usefulness of acoustic radiation force impulse (ARFI) elastography for noninvasive evaluation of liver fibrosis in rats.Methods A total of 70 male Wistar rats were included in the group for dimethylnitrosamine (DMN)-induced liver injury,and 10 saline-injected rats were used as normal control.Hepatic injury was induced by a single intraperitoneal injection of DMN at a dose of 50 mg/kg of body weight.Several rats in the group with DNM injected and the normal control group were randomly selected and sacrificed at each of the following post-injection time:day 5,7,10,14,21,24,and 28.And their livers were taken for pathology analysis.All the rats underwent ARFI elastography before sacrificed in order to acquire a shear wave velocity (Vs) to represent liver stiffness.Correlation between Vs and the histological finding was analysed.ResultsAmong 58 successfully modeled rats,9,13,14 and 12 rats were found to be with S1,S2,S3 and S4 of liver fibrosis pathologically,respectively.And 10 rats were found to be with severe inflammatory activity without any fibrosis.Values of Vs increased with the stage of liver fibrosis ( P ＜0.05).There was a significant correlation between Vs and stage of liver fibrosis ( r =0.947,P =0.000).The areas under ROC curve for the diagnosis of fibrosis S≥S1,S≥S2,S≥S3 and S=S4 were 0.983,0.995,0.999 and 0.964,respectively;for the cutoff values of Vs were 1.59 m/s,2.13 m/s,2.33 m/s and 2.51 m/s,respectively,the sensitivity was 95.8％,92.3％,100％ and 84.6％,and specificity was 100％,100％,96.9％ and 95.6％,respectively.The values of Vs in the group with severe inflammatory activity were significantly higher than those in the control group ( P =0.000).ConclusionsARFI has a relatively high value in the evaluation of liver fibrfosis in rats,while severe inflammatory activity may affect its accuracy.

In order to improve tensile property of vascular scaffold, we blended silk fibroin with novel human-like collagen with the mass ratio of 9:1, 7:3 and 5:5 (W/W), and then fabricated blood vessel tubular graft by freeze-drying process. We studied microstructure, mechanical properties, elements composites, degradability and biocompatibility of vascular scaffolds. These results showed that tubular scaffold with mass ratio 7:3 exhibited interconnected porous structure with pore size at (60 +/- 5) microm and porosity of 85%; achieved the desirable mechanical property (strain of 50% +/- 5% and stress of 332 +/- 16 kPa); had relatively slow degradation rate; could enhance cell adhesion and proliferation and had superior biocompatibility.
Biocompatible Materials ; chemistry ; Biomechanical Phenomena ; Blood Vessels ; physiology ; Collagen ; chemistry ; Fibroins ; chemistry ; Humans ; Materials Testing ; Porosity ; Tissue Engineering ; methods ; Tissue Scaffolds

Objective To investigate the effectsof adipocyte-specific Structural maintenance of chromosomes 5(Smc5)knockouton glucose and lipid metabolism in mice.Methods Adipocyte-specific Smc5 knockout mice(AKO mice)were constructed based onclustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9(CRISPR/Cas9)systems,and Smc5flox/flox mice(Flox mice)were used as controls.The Smc5 knockout efficiency of adipose tissue in mice was verified by qRT-PCR and Western blot.The body fat content was detected by dual energy X-ray absorptiometry(DEXA).The morphology of adipose tissue was observed by hematoxylin-eosin staining and the area distribution of adipocytes was calculated.TG,HDL-C,LDL-C,free fatty acid(FFA),intraperitoneal glucose tolerance test(IPGTT)and intraperitoneal insulin tolerance test(IPITT)were compared.Results The AKO mouse model of fat specific knockout of Smc5 gene was successfully constructed.Smc5 mRNA in groin fat(iWAT),epididymal fat(eWAT)and brown adipocyte(BAT)of AKO mice,body weight after 15 weeks,organ weight of iWAT at 31 weeks,organ weight of eWAT,organ weight of BAT,body weight,fat mass,fat mass/weight,LDL-C,and percentage of 15-minute blood glucose in IPITT were lower than those of Flox mice(P<0.05 or P<0.01).Conclusions Adipocyte-specific Smc5 gene knockout improves glucose and lipid metabolism by affecting adipose tissue production.

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.

Aiming at comparing the pre-operative and post-operative gait characteristics and therefore establishing post-operative rehabilitation guidance for patients with end-stage knee osteoarthritis (KOA) merged with varus deformity, this study captured the level walking and sit-to-stand trials of 9 patients with 3-dimensional motion analysis system and after which musculoskeletal multi-body dynamic analysis was conducted. The study indicated that the average range of motion (ROM) of the proposed-surgical knee was 24.4°-57.6° and that of the non-surgical knee was 22.5°-71.5°. The knee ROM of control group during level walking was 7.2°-62.4°. When the unilateral KOA patients stood up from chair to complete the sit-to-stand movement, the ground reaction forces (GRFs) symmetry was 0.72-0.85, which means that the non-surgical limb bear the majority of body weight. The GRFs of the bilateral KOA patients were smallest during the sit-to-stand movement. The strategy that the non-surgical limb dominates in loading bearing taken by the unilateral KOA patients to cover most post-operative daily activities could increase the risk of KOA among non-surgical side joints as a result of long-term excessive loading-bearing. The study, on kinematics and biomechanical characteristics of patients with KOA merged with varus deformity, could help to understand the pathogenesis of KOA merged with varus deformity from the perspective of biomechanics and to provide strong clinic guidance for the pre-operative evaluation, prevention and post-operative recovery for patients.