Objective To establish a genetic monitoring method for laboratory quails.Methods Quail microsatellite loci were searched in the literature,and microsatellite DNA loci suitable for quails were screened by an interspecific transfer method in closely related species,namely chickens and ducks.Quail liver DNA was extracted as a template,and the corresponding loci were screened by PCR amplification and agarose gel electrophoresis.On the basis of amplification of the selected microsatellite loci,the number of alleles,polymorphisms,and microsatellite loci combinations for quail genetic quality detection were selected and detection method were developed.Results We preliminary determined 23 microsatellite loci for genetic monitoring of closed-colony laboratory quails.Conclusions A genetic monitoring method for laboratory quails was preliminary developed.

OBJECTIVE To explore the anti-liver fibrosis effect and mechanism of Atractylenolide Ⅲ-induced hepatic stellate cell(HSC)senescence.METHODS ASCT2 siRNA and Atractylenolide Ⅲ(40 μmol·L-1)acted on human hepatic stellate cells LX2 respectively to inhibit ASCT2,MTT was used to evaluate cell viability,EdU method was used to detect cell proliferation,and se-nescence associated-β-galactosidase(SA-β-Gal)staining was used to detect cell senescence;Western blot was used to detect chan-ges in the LC3-Ⅱ/Ⅰ ratio in LX2 cells,laser confocal detection was used to detect changes in LC3 autophagy flow and error protein accumulation,and the fluorescence of the lysosomal marker LAMP1 was also observed to detect lysosomal function and quantity;kits were applied to detect ROS and MDA levels as well as SOD activity in LX2 cells,and flow cytometry was used to analyze mitochondrial ROS levels and membrane potential.A CCl4-induced mouse liver fibrosis model was constructed.Atractylenolide Ⅲ was administered at 20,30,or 40 mg·kg-1.HE,Masson,and Sirius Red staining were used to observe liver tissue damage and collagen deposition.Western blot was used to detect the expression levels of P21 and P16 in mice in each group,and SA-β-Gal staining and immunohistochemistry were used to analyze the situation and origin of senescent cells.RESULTS After inhibiting ASCT2,the viabil-ity of LX2 cells decreased and senescence increased(P<0.01).Meanwhile,the autophagy function was enhanced and the number of lysosomes was increased but the function was weakened.After adding chloroquine(CQ)to clear lysosomes,the cell viability and auto-phagy function increased(P<0.01).After inhibiting ASCT2,the levels of MDA and ROS in LX2 cells increased,and the activity of SOD decreased(P<0.01).Among them,the level of mitochondrial ROS increased and the membrane potential decreased(P<0.01).After adding rotenone,the cellular redox homeostasis was improved,and the number of lysosomes was restored(P<0.01).In vivo experimental results showed that compared with the model group,Atractylenolide Ⅲ improved liver tissue structural damage and collagen deposition,induced HSC senescence in liver tissue of mice with liver fibrosis,and inhibited HSC activation marker α-smooth muscle actin(α-SMA),promoted the expression of senescence indicators P16 and P21(P<0.01).CONCLUSION Atractylenol-ide Ⅲ induces an increase in mitochondrial ROS and a decrease in membrane potential by inhibiting ASCT2,which further promotes the enhancement of HSC autophagy function,increases the number of lysosomes and weakens their function,thereby inducing the se-nescence of activated HSCs.

Objective To explore the effect of a phased rehabilitation training programme to relieve shoulder dys-function in patients after neck dissection and to provide effective solutions for postoperative shoulder joint function recov-ery of patients.Methods This study has been reviewed and approved by the Ethics Committee,and informed consent has been obtained from patients.A phased rehabilitaiton training programme for the shoulder after neck dessection was developed through literature review and discussion,and 70 eligible patients from Hospital of Stomatology,Sun Yat-sen University from December 2020 to April 2021 were selected and randomly divided into the test group and control group(35 patients in each group).The control group underwent motor rehabilitation training from 6 weeks postoperative to 1 year after surgery,such as shoulder mobility and coordination training and small range of motion training of the neck,while the test group took part in a rehabilitation training program that included familiarization maneuver training,protective rehabilitation,exercise rehabilitation,and resistance training in the following four stages:preoperative,postop-erative general anesthesia and awake until the removal of stitches,the removal of stitches until 6 weeks after surgery,and 6 weeks after surgery until 1 year after surgery.The frequency of training in both groups was at least 3 days per week,and the length of each training session was 10-15 min.The intensity of exercise was 2-3 points on the Borg Conscious Ex-ercise Intensity Scale(i.e.,mild-to-moderate tachypnea or fatigue).The neck dissection injury index(NDII)was used to evaluate the quality of life related to shoulder joint function at four time points:preoperative,postoperative 3 months,postoperative 6 months,and postoperative 12 months.The higher the score,the better the quality of life.Results 28 cases in the test group and 32 cases in the control group completed a one-year follow-up.At 3 and 6 months postopera-tive,the NDII of the test group was significantly higher than that of the control group[3 months postoperative:test group(93.48±9.36)vs.control group(80.00±11.34)(P＜0.001),6 months postoperative:test group(98.21±4.76)vs.control group(90.70±9.12)(P＜0.001)];12 months after surgery,the NDII of the test group(97.23±4.88)was still higher than that of the control group(96.33±4.49),but the difference was not statistically significant(P=0.458).The difference in NDII scores among subjects at 3,6,and 12 months after surgery was statistically significant in each group(P＜0.001).Conclusion The application of the phased rehabilitation training method in neck dissection patients has a feasibility and could improve the quality of life of patients'shoulder joint function within 6 months after surgery.

Objective To systematically evaluate the risk factors for new-onset atrial fibrillation after off-pump coronary artery bypass grafting (OPCABG). Methods PubMed, EMbase, The Cochrane Library, CNKI, Wanfang, VIP, SinoMed were searched to collect published literature on risk factors for new-onset atrial fibrillation after OPCABG from inception to September 2022. Two authors independently screened, extracted data and evaluated the quality. The Newcastle-Ottawa Scale (NOS) was used to evaluate the quality of the included studies, and Stata 12.0 and RevMan 5.4 softwares were used for meta-analysis. Results A total of 18 researches were included, including 6 354 patients of OPCABG. The NOS scores of the included studies were 6-8 points. Meta-analysis showed that age [MD=2.56, 95%CI (1.61, 3.52), P<0.001], hypertension [OR=1.77, 95%CI (1.18, 2.66), P<0.001], EuroSCORE Ⅱ score [MD=0.70, 95%CI (0.34, 1.06), P<0.001], frequent atrial premature beats or atrial tachycardia [OR=3.77, 95%CI (2.13, 6.68), P<0.001], left atrium diameter (LAD) [MD=1.64, 95%CI (0.26, 3.03), P=0.010], left ventricular ejection fraction (LVEF) [MD=−1.84, 95%CI (−2.85, −0.83), P<0.001], right coronary stenosis [OR=2.49, 95%CI (1.29, 4.81), P=0.006], three-vessel coronary artery lesions [OR=0.73, 95%CI (0.54, 0.97), P=0.030], not using β blockers [OR=0.81, 95%CI (0.69, 0.96), P=0.010], operation time [MD=10.13, 95%CI (8.15, 12.10), P<0.001], duration of mechanical ventilation [OR=2.85, 95%CI (1.79, 3.91), P<0.001] were risk factors for new-onset atrial fibrillation after OPCABG. Conclusion Advanced age, hypertension, high EuroSCOREⅡ score, frequent atrial premature beats or atrial tachycardia, increased LAD, decreased LVEF, right coronary stenosis, three-vessel coronary artery lesions, not using β blockers, prolonged operation time and mechanical ventilation are risk factors for new-onset atrial fibrillation after OPCABG. Due to factors such as the methodology, content and quality of the included literature, the conclusion of this study need to be supported by more high-quality studies.

Objective To elucidate the clinical significance in facilitating timely diagnosis and treatment of RMPP in children by investigating the association between infection caused by macrolide-resistant Mycoplasma pneu-moniae(MRMP)and refractory Mycoplasma pneumoniae pneumonia(RMPP)in pediatric patients.Methods The clinical data of 714 hospitalized children with refractory Mycoplasma pneumoniae pneumonia(RMPP)were retro-spectively analyzed.Bronchoscopy and bronchoalveolar lavage fluid(BALF)were performed on each subject,and the BALFs were collected to detect mutation sites in the V region of 23S rRNA for Mycoplasma pneumoniae DNA.Based on the gene detection results,children with RMPP were categorized into a macrolide-resistant group and a control group(non-macrolide-resistant group).Results A total of 714 children diagnosed with refractory Mycoplasma pneumoniae pneumonia(RMPP)were enrolled in this study,including 509 cases in the macrolide-resistant group and 205 cases in the control group.Among them,there were 369 males(54.7%)and 345 females(45.3%).The macrolide-resistant group exhibited higher average age,fever duration,and hospitalization days compared to the control group.Furthermore,elevated levels of white blood cell count(WBC),neutrophil percentage(NE%),high-sensitivity C-reactive protein(hs-CRP),lactate dehydrogenase(LDH),and interleukin-6(IL-6)were observed in the macrolide-resistant group when compared to the control group(P<0.05 or P<0.000 1).Compared to the control group,children with macrolide-resistant Mycoplasma pneumoniae pneumonia(RMPP)exhibited higher incidences of lung consolidation,pleural effusion,necrotic pneumonic lesions,severe MPP(SMPP)/fulminant MPP(FMPP),flocculent and viscous tracheal secretions,severe mucosal lesions(erosion,ulceration or necrosis),bronchial inflammatory stenosis,endo-bronchial plastic phlegm plugs and extra-pulmonary complications(P<0.05 or P<0.0001).Conclusions MRMP infection can contribute to the development of RMPP,potentially exacerbating respi-ratory conditions in affected children.Timely bronchoscopy and collection of BALF samples for accurate evaluation of respiratory tract lesions and detection of MRMP infection have significant implications for guiding precise clinical diagnosis and treatment.

Objective To study the changes of blood glutathione peroxidase (GSH-px),superoxide dis-mutase (SOD),malondialdehyde (MDA) and pathological tissues in the rat contrast-induced nephropathy (CIN) model,and to determine the role of oxidation mechanism in CIN.Methods A total of 40 adult male SD rats were selected and divided into three big groups and five small groups.After constructing the model,six rats with good status were taken from each group for conducting the experiment.The serum GSH-px,SOD and MDA levels were measured,the renal tissue biopsy was performed and the morphological changes of kid-ney cells were compared.Results There was no statistically significant difference in the baseline data among the blank control group,the control group and the experimental group (P>0.05).There was no statistically significant difference in serum GSH-px,SOD and MDA levels before model construction,at 24,48 h after model construction between the blank control group and the control group (P>0.05).There were statistical-ly significant differences in serum GSH-px,SOD and MDA levels of the experimental group between before model construction and after model construction (P<0.05).There was no statistically significant difference in serum GSH-px,SOD and MDA level in the experimental group between at 24 h after modelling and 48 h af-ter modeling (P>0.05).There was no statistically significant difference in serum GSH-px,SOD and MDA levels at 24 h after modeling among the three groups (P>0.05).There were statistically significant differ-ences in serum GSH-px,SOD and MDA levels at 48 h after modeling among the three groups and their pairs (P<0.05).The pathological sections of the blank control group and control group showed no obvious abnor-mal changes in glomeruli,renal tubule and renal interstitium.Renal interstitial fibrosis and inflammatory cell infiltration were seen after 24 h in the experimental group,but there was no obvious change in the renal tu-bules.After 48 h,moderate focal-like atrophy of renal tubules,epithelial cell granule degeneration and vacuolar changes were obviously seen.Conclusion The oxidative stress mechanism plays a role in CIN.The contrast a-gent acute renal injury mainly acts on the renal tubules and renal interstitium,and there is no obvious damage to the glomeruli.

Humans ; Melanoma/therapy* ; Skin Neoplasms/therapy* ; Methylation ; Prognosis ; Immunotherapy ; RNA ; Biomarkers, Tumor ; Melanoma, Cutaneous Malignant

Objective:To investigate the value of preoperative MRI measurements of hamstring (semitendinosus+gracilis) tendon cross-sectional area in predicting intraoperative graft diameter for anterior cruciate ligament reconstruction (ACLR).Methods:A total of 265 preoperative MRI were retrospectively collected in the Third Affiliated Hospital of Southern Medical University from January 2013 to August 2020 for patients who underwent single-bundle ACLR of hamstring tendon. Patients were divided into a graft diameter≥8 mm group (129 patients) and a graft diameter<8 mm group (136 patients) according to intraoperative graft diameter. The cross-sectional areas of the semitendinosus and gracilis tendons were measured at the largest level of the femoral condyle on preoperative MRI cross-sectional images, and the two were summed to obtain the cross-sectional area of the hamstring tendon. The independent samples t-test was used to compare the differences in the cross-sectional area of each tendon between the graft diameter≥8 mm group and the graft diameter<8 mm group. The Spearman correlation analysis was used to assess the correlation between tendon cross-sectional area and intraoperative graft diameter. Multi-factor logistic regression analysis was used to screen the influence of tendon cross-sectional area on intraoperative graft diameter. The effectiveness of intraoperative graft diameter≥8 mm was assessed by plotting the receiver operating characteristic curves. Results:Intraoperative measurement of graft diameter was 7.5 (7.5, 8.0) mm. The cross-sectional area of the popliteal tendon was (21.4±4.6) mm 2 in the graft diameter≥8 mm group and (15.6±3.7) mm 2 in the graft diameter<8 mm group. Statistically significant differences were found in the cross-sectional areas of the semitendinosus, soleus and hamstring tendons between the graft diameter≥8 mm group and graft diameter<8 mm group ( t=-10.26, -10.29, -11.47, P<0.001). Intraoperative graft diameter was positively correlated with the cross-sectional area of the semitendinosus, gracilis, and hamstring, with correlation coefficients of 0.57 ( P<0.001), 0.58 ( P<0.001), and 0.62 ( P<0.001), respectively. Multi-factor logistic regression showed that popliteal tendon cross-sectional area was a predictor of intraoperative graft diameter (OR=1.45, 95%CI 1.32-1.59, P<0.001). The area under the curve for popliteal tendon cross-sectional area to predict intraoperative graft diameter≥8 mm was 0.838 (95%CI 0.792-0.885), with a critical value of 20.0 mm 2, a sensitivity of 0.581, and a specificity of 0.941. Conclusion:The measurement of the cross-sectional area of the hamstring muscle on preoperative MRI can predict the diameter of the autologous hamstring graft of ACLR.

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease affecting both upper and lower motor neurons (MNs) with large unmet medical needs. Multiple pathological mechanisms are considered to contribute to the progression of ALS, including neuronal oxidative stress and mitochondrial dysfunction. Honokiol (HNK) has been reported to exert therapeutic effects in several neurologic disease models including ischemia stroke, Alzheimer's disease and Parkinson's disease. Here we found that honokiol also exhibited protective effects in ALS disease models both in vitro and in vivo. Honokiol improved the viability of NSC-34 motor neuron-like cells that expressed the mutant G93A SOD1 proteins (SOD1-G93A cells for short). Mechanistical studies revealed that honokiol alleviated cellular oxidative stress by enhancing glutathione (GSH) synthesis and activating the nuclear factor erythroid 2-related factor 2 (NRF2)-antioxidant response element (ARE) pathway. Also, honokiol improved both mitochondrial function and morphology via fine-tuning mitochondrial dynamics in SOD1-G93A cells. Importantly, honokiol extended the lifespan of the SOD1-G93A transgenic mice and improved the motor function. The improvement of antioxidant capacity and mitochondrial function was further confirmed in the spinal cord and gastrocnemius muscle in mice. Overall, honokiol showed promising preclinical potential as a multiple target drug for ALS treatment.

OBJECT IVE To evaluate the application effect of the whole cour se medication management mode led by pharmacists in rheumatic immune diseases. METHODS A total of 122 patients treated with tacrolimus or cyclosporine in the department of rheumatology and immunology of Wuhan No. 1 Hospital from 2018 to 2020 were selected as the study subjects. Among them ，44 cases in the control group were under the traditional supervision mode ；78 patients in the observation group adopted the whole course medication management mode led by pharmacists ，that was ，individual pharmacists and specialist clinical pharmacists cooperated and led ，and not only participated in the whole process of drug treatment but also involved in the whole process of therapeutic drug monitoring （TDM）. On the basis of the control group ，the division of labor and cooperation among medical，pharmaceutical and nursing parties were strengthened ，and the homogeneous supervision was carried out for the outpatients and inpatients from admission to discharge . The daily dose of medication ，the rate of reaching the standard of blood drug concentration ，the incidence of problematic samples （the sample was calculated by the number of times ），the average hospitalization days ，the re-admission rate within 6 months after discharge ，the medication compliance score and the patient ’s satisfaction rate were compared between the two groups. RESULTS In the control group ，53 times of TDM were performed ， including 18 times of tacrolimus monitoring and 35 times of cyclosporine monitoring ；in the observation group ，123 timesof TDM were performed ，including 55 times of tacrolimus monitoring and 68 times of cyclosporine monitoring. The daily dose of tacrolimus ，the daily dose of cyclosporine ，the rate of reaching the standard of cyclosporine blood drugconcentration，the inc idence of problematic samples ，the rate of re-admission within 6 months after discharge ， the medication compliance score and the patient ’s satisfaction rate in the observation group were significantly better than those in the control group （P＜0.05）. CONCLUSIONS It can effectively improve the effect of the quality of pharmaceutical care to implement whole course and homogeneous medication management led by pharmacists and provide precise drug guidance for patients with rheumatic and immune diseases.