This study aims to prepare nanobodies for the epitopes of spike protein(S)of porcine ep-idemic diarrhea virus(PEDV),and verify its reactivity.The expression vector pCZN1-SN was con-structed by the prokaryotic expression system,and SN fragment was expressed in prokaryotic ex-pression and purified by Ni-NTA chromatography column.The SN nanobodies were displayed u-sing phage display technology and screened from the natural nanobody library.The results showed that SN fragments with a size of 26 kDa were obtained by prokaryotic induction,and the specific nanobodies were obtained through three rounds screening by phage display technology.One strain with the best reactivity was selected for prokaryotic expression and purified.The nanobodies were obtained with a size of 14 kDa by Western blot and demonstrated to have a good binding ability to PEDV SN protein.In summary,nanobodies with epitope fragments of PEDV S protein were suc-cessfully screened and prepared based on the phage display technology,which provided a new way for the in-depth application of nanobodies.

Objective To investigate the biological effects of Pasteurella multocida(Pm)culture supernatant and Pm-derived outer membrane vesicle(OMV)on bladder cancer cells.Methods Pm was cultured and its supernatant was collected.The effects of the supernatant on proliferation,migration and invasion of bladder cancer cell lines(T24 and 5637)were assessed by cell counting kit 8(CCK-8),wound healing assay,and Transwell migration and invasion assays with phosphate-buffered saline(PBS)and brain heart infusion(BHI)broth as controls.Pm-OMV were isolated from the supernatant via ultracentrifugation,and the remaining components of the supernatant served as control.The effects of Pm-OMV on proliferation,migration and invasion of T24 and 5637 cells were assessed by CCK-8 and Transwell migration and invasion assays.Apoptosis was analyzed by flow cytometry.A nude mouse xenograft tumor model was established.After intratumoral multi-point injections of Pm-OMV or PBS,the tumor growth was evaluated and the effects of Pm-OMV on proliferation and apoptosis of bladder cancer cells in vivo were verified by Ki67(a proliferation marker)immunohistochemical staining and TUNEL assay.Results Pm culture supernatant significantly inhibited the proliferation,invasion,and migration of T24 and 5637 cells in vitro compared with PBS and BHI controls(all P＜0.01).Pm-OMV not only inhibited the proliferation,invasion,and migration of T24 and 5637 cells,but also induced the apoptosis,and the differences were significant compared with the remaining components of the supernatant(all P＜0.05).The nude mouse subcutaneous tumor transplantation experiment further confirmed that Pm-OMV inhibited the proliferation of bladder cancer cells and promoted apoptosis in vivo,and the differences were significant compared with the PBS control(all P＜0.05).Conclusion Pm-OMV can inhibit the proliferation,invasion,and migration of bladder cancer cells and promote the apoptosis.It provides an experimental basis for studying the mechanism of microbial regulation of tumor progression and for developing new treatment strategies for bladder cancer.

This study aims to prepare nanobodies for the epitopes of spike protein(S)of porcine ep-idemic diarrhea virus(PEDV),and verify its reactivity.The expression vector pCZN1-SN was con-structed by the prokaryotic expression system,and SN fragment was expressed in prokaryotic ex-pression and purified by Ni-NTA chromatography column.The SN nanobodies were displayed u-sing phage display technology and screened from the natural nanobody library.The results showed that SN fragments with a size of 26 kDa were obtained by prokaryotic induction,and the specific nanobodies were obtained through three rounds screening by phage display technology.One strain with the best reactivity was selected for prokaryotic expression and purified.The nanobodies were obtained with a size of 14 kDa by Western blot and demonstrated to have a good binding ability to PEDV SN protein.In summary,nanobodies with epitope fragments of PEDV S protein were suc-cessfully screened and prepared based on the phage display technology,which provided a new way for the in-depth application of nanobodies.

OBJECTIVE:Knee adduction moment and knee adduction angular impulse enlargement is the main biomechanical risk factor of knee osteoarthritis.According to the survey,a change in the foot progression angle could effectively change the motion mode of patients with knee osteoarthritis.However,the impact of toe-in and toe-out on knee adduction moment and knee adduction angular impulse in young and elderly patients did not reach a consensus.Therefore,this study comprehensively discussed the effect of foot progression angle on knee adduction moment and knee adduction angular impulse in different populations through meta-analysis and provided a reference for the treatment of knee osteoarthritis. METHODS:By June 2022,searches were conducted on Web of Science,EBSCO,PubMed and CNKI databases using"foot progression angle,knee adduction moment,knee adduction angular impulse,gait"as Chinese and English search terms.Self-controlled randomized controlled studies analyzing the effects of toe-in and toe-out on knee adduction moment bimodality and knee adduction angular impulse were included.The cochrane bias risk assessment tool was utilized to make a quality evaluation of the literature.Stata 15.1 software was used for subgroup analysis to determine the effect of foot progression angle on knee adduction moment and knee adduction angular impulse.Meta-regression analysis was used to further determine characteristics of outcome indicators(knee adduction moment,knee adduction angular impulse)changing with foot progression angle. RESULTS:(1)A total of 15 self-control trials and 2 randomized controlled trials(455 subjects)were included in the meta-analysis.All of the included articles were of medium to high quality.(2)The meta-analysis results showed that the toe-in gait could reduce the first peak of knee adduction moment(SMD=-0.380,95%CI:-0.710 to-0.060,P=0.022)and knee adduction angular impulse(SMD=-1.470,95%CI:-2.160 to-0.770,P<0.001)in young patients.The toe-out gait reduced the second peak of knee adduction moment(SMD=-0.720,95%CI:-1.010 to-1.440,P<0.001)in young patients.In addition,toe-in gait could reduce the first peak of knee adduction moment in elder patients(SMD=-0.550,95%CI:-0.800 to-0.300,P<0.001),but increase the second peak knee adduction moment of elderly(SMD=0.280,95%CI:-0.010 to 0.560,P=0.047).The toe-out gait could decrease the second peak knee adduction moment in this population(SMD=-0.510,95%CI:-0.830 to-0.190,P=0.002).(3)Meta-regression showed that the greater the toe-out in elderly patients,the lower the second peak knee adduction moment. CONCLUSION:(1)Toe-in reduced the first peak knee adduction moment and knee adduction angular impulse in young knee osteoarthritis patients aged 18 to 34 years.Since knee adduction moment and knee adduction angular impulse were associated with medial knee loading and knee osteoarthritis incidence,toe-in gait intervention may be a suitable rehabilitation strategy for young patients.(2)Toe-in increased the second peak of knee adduction moment in older knee osteoarthritis patients over 60 years of age,which may exacerbate knee osteoarthritis in this population.However,the second peak of knee adduction moment during walking in this population decreases as the toe-out increases,contributing to a reduction in medial knee loading,suggesting that older patients may consider using toe-out gait during walking.

BACKGROUND:Delayed onset muscle soreness is closely related to skeletal muscle micro-injury, but the exact mechanism of skeletal muscle micro-injury is not yet clear. OBJECTIVE:To observe the histomorphological and ultrastructure changes of skeletal muscle micro-injury models induced by eccentric exercise. METHODS: Forty male Sprague-Dawley rats were randomly divided into quiet control group, immediately after exercise group, post-exercise 24 hours group, post-exercise 48 hours group and post-exercise 72 hours group. In the latter four groups, the rats were subjected to intermittent running on the-16° slope at a speed of 16 m/min: 5 minutes movement, 2 minutes rest and totaly 120 minutes. Rats in the latter four groups were observed immediately, at 24, 48, 72 hours after exercise. RESULTS AND CONCLUSION:After eccentric exercise, the morphology and ultrastructure of rat’s skeletal muscle were both changed to different extents, and Desmin and Vimentin were dyed off for anti-desmin antibody staining at varying degrees. It indicates that one-time eccentric exercise can cause delayed skeletal muscle micro-injury.

Objective To study the effects of Ginkgo biloba extract (GBE) on free radical metabolism of different fiber types of quadriceps in exercised rats and its mechanism. Method Increasing load exercised rat model was selected. The test rats were orally given GBE aqueous suspension (100 mg/ kg?d), and the controls with water. The levels of malondialdehyde (MDA) and the activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px) of red muscle, white muscle and whole quadriceps of exercised rats were examined before exhaustive exercise, at instant,24h and 48h after strenuous exhaustive exercise. Results Compared with the controls,MDA contents were 49.69% higher in white muscle (P

Objective:Study of intensive endurance training and supplementation of herbal polysaccharide extracts on levels of IL-2, IL-4,IL-10 in serum of rats.Methods:120 Wistar Rats were randomly divided into 6 groups: sedentary control group(n=20), sedentary with supplementation of APS group(n=20), sedentary with supplementation of ABPS group(n=20), training group(n=20), training with supplementation of APS group(n=20) and training with supplementation of ABPS group(n=20). After 4-week and 6-week swimming, IL-2, IL-4,IL-10 in serum of rats and supernatant of PBMC were tested through ELISA.Results:After 6 weeks training, (1) In training group, the level of IL-2 in serum was lower than that in sedentary control group(P

In order to probe into the peripheral mechanism of exercise-induced low level of serum testosterone and to explore an effective Chinese medicine compound for ameliorating this serious situation, a rat model of intensive swimming training was conducted, and the ultrastructure of testis in rats and the effects of Chinese medicine compound on this were studied with morphometry. Results showed that the volume of lipid droplets in rat testicular Leydig cells decreased after long term endurance training,which may be one of the causes of exercise-induced low level of serum testosterone; however, the administration of traditional Chinese medicine compound for invigorating qi and tonifying the kidney could improve the transportation of cholesterol and recovery of the testosterone level in serum.

Objective: To investigate the effects of soy hydrolysed peptides supplementation on small intestinal morphology and nitrogen absorption in rats,and illustrate the characteristic of absorption on soy hydro? lysed peptides supplementation in small intestinal. Methods: 30 male SD rats were randomly divided into three groups: Placebo group (water supplemented, Pla group); Isolated soy protein-supplemented group (Pro group); Soy hydrolysed peptides-supplemented group (Pep group). After one week meals adaptation, Metabolic test had done, small intestinal morphology, and the activity of aminopeptidase were determined. Results: (2) The content of intestinal epithelial protein in Pep group increased 48.60% than Pro group, and increased 91.37% than Pla group (P

Objective: We performed a randomized double-blind study in distance runners to investigate the possible biological effects of daily consumption of soybean peptides. Methods: Twenty-one trained male distance runners, randomly divided into three groups: control group (C,n=7), carbohydrate supplementation group (CS, n=6) and soybean peptides supplementation group (SP,n=8), took part in the study.Their hormonal and serum metabolic profile was examined in a 4-week program of strenuous training. Daily training (4～6 hours per day, endurance and strength), food and medication were similar for all athletes. Group SP received a soy sports beverage supplement containing 8g soybean peptides and 35g carbohydrate once a day. Group CS received a sport drink containing 35g carbohydrate. Group C supplemented a placebo identical in appearance and flavor. Body weight, lean body mass, RPE, serum total testosterone, blood urea, hemogram, lactate dehydrogenase, creatine kinase and immunoglobulins were measured before, during and after the 4-week training program. Results: Data demonstrated that Group SP had an increase in body weight, especially in lean body mass and cyclic total testosterone level (P