Objective To observe the expression changes of inflammatory cytokines of human tendon-derived stem cells (TDSCs) under uniaxial stretching.Methods TDSCs were isolated from human gracilis tendon by collagenase digestion at very low density.Cells were detected for surface markers by flow cytometry,and tested for multi-differentiation potential in vitro.Repetitive uniaxial stretching was applied on the cells at 0％,4％,8％ or 10％ strain.Under the microscopy,cell alignment was observed after mechanical stretching.Expressions of inflammation factors COX-2 and MMP-1 were detected by qPCR and western blotting.Results TDSCs were successfully isolated from human gracilis tendon.Inflammatory cytokines CD29,CD44 and CD105 were positive,but CD45 and CD14 were negative.TDSCs could differentiate into osteocytes,adipocytes and chondrocytes in vitro.Cells were not realigned4 hours after mechanical stretching.MMP-1 mRNA expression was significantly down-regulated at 4％ strain (0.090 ± 0.007) compared to that at 0％ strain (0.247 ± 0.032,P ＜ 0.05).No significant difference was found in COX-2 mRNA expression at 4％ and 8％ strain (both was 0.005 ±0.001,P ＞0.05).MMP-1 and COX-2 mRNA expressions at 8％ strain (0.168 ± 0.040 and 0.007 ± 0.001)revealed no significant differences from those at 0％ strain (0.134 ±0.075 and 0.006 ±0.003) (P ＞0.05),whereas at 10％ strain MMP-1 and COX-2 mRNA expressions were significantly up-regulated (0.047 ± 0.003 and 0.496 ± 0.036) compared to those at 0％ strain (0.011 ± 0.003 and 0.005 ± 0.003)(P ＜ 0.05).Changes in MMP-1 and COX-2 protein expressions revealed similar trend as their mRNA expressions.In contrast to the setting of 0％ strain,4％ strain induced down-regulated MMP-1 and COX-2 proteins,8％ strain induced no significant changes in MMP-1 and COX-2 proteins,and 10％ strain induced up-regulated COX-2 protein despite minor increase in MMP-1 protein.Conclusions Mechanical stretching can affect the secretion of inflammatory cytokines.Low tensile stretch is associated with decreased expression of inflammatory cytokines while high tensile stretch promotes secretion of inflammatory cytokines.

Objective To explore the influence of social support and coping style on body and mind symptoms of patients with laryngeal carcinoma. Methods SCL-90, SSRS and MCMQ were used for the questionnaire survey of 168 patients with laryngeal carcinoma. Results The result of SCL-90 showed that all items were higher than those of norm in China, except the interpersonal relationship sensitivity and psychotic symptoms. The result of SSRS showed that the total score of social support was higher than that of norm, while the availability of support was obviously lower than that of norm. The result of MCMQ showed that avoidance and submissiveness were used frequently in patients, and avoidance was negatively correlat-ed with the factors of psychology. Conclusions Before the nursing plan and health consultation are set for patients with laryngeal carcinoma, the patients' social support should be used sufficiently, and the effect of coping style on patients' mental health should be evaluated.

Objective:To explore the correlation between fibular head height and varus knee osteoarthritis occurrence and severity.Methods:A retrospective analysis was performed on 618 participants (618 knees, 184 males and 434 females, mean age 61.12±10.98 years) who underwent standard weight-bearing full-leg radiographs and were diagnosed as non-knee osteoarthritis or varus knee osteoarthritis from January 2019 to June 2019. Knee osteoarthritis was diagnosed according to Kellgren-Lawrence grading: 0-I grades were diagnosed as non-osteoarthritis, II-IV grades were diagnosed as osteoarthritis. Joint line convergence angle (JLCA), medial proximal tibial angle (MPTA) and hip-knee-ankle angle were measured on X-rays to reflect varus deformity. The fibular head height was defined as the vertical distance from upper edge of fibular head to lateral tibial plateau. Patients were divided into 5 groups according to Kellgren-Lawrence grading. Differences of age, gender, height, weight, body mass index, varus deformity (JLCA, MPTA and hip-knee-ankle angle) between Kellgren-Lawrence 0-IV grades were compared. Ordinal logistic regression was performed to analyze the correlation between fibular head height and Kellgren-Lawrence grades. Pearson's correlation analysis was used for the correlation among fibular head height, JLCA, MPTA and hip-knee-ankle angle, and the main factor of JLCA, MPTA and hip-knee-ankle angle was extracted by factor analysis. Multiple linear regressions were used to analyze the correlation between fibular head height and varus deformity score.Results:There were 68, 66, 97, 98, 289 participants in Kellgren-Lawrence grades 0-IV respectively that was 134 participants were diagnosed as non-osteoarthritis and 484 participants were diagnosed as osteoarthritis. Fibular head height and MPTA showed a decreasing trend ( F=129.076, 24.875; P<0.001) while JLCA and hip-knee-ankle angle showed an increasing trend ( F=414.346, 105.996; P<0.001) with the increase in Kellgren-Lawrence grading. Age, body mass index and fibular head height are influencing factors of Kellgren-Lawrence grading with OR(95%CI) were 1.116(1.093, 1.141), 1.363(1.060, 1.754), 0.617(0.575, 0.662) . Fibular head height was negatively correlated with JLCA and hip-knee-ankle angle ( r=-0.641, -0.478; P<0.001) , respectively, and positively correlated with MPTA ( r=0.320, P<0.001). There were significant correlations between age, fibular head height and the varus deformity score ( β=0.274, -0.457; P<0.001). Conclusion:Fibular head height of patients with varus knee osteoarthritis is lower than that of non-osteoarthritis. In addition to age and body mass index, fibular head height is a risk factor for varus knee osteoarthritis occurrence. The smaller the fibular head height is, the more serious the osteoarthritis severity and varus deformity are.

OBJECTIVE：To study the protective effects of Scutellaria amoena enthanol extract and its different solvent parts on liver injury induced by CCl 4. METHODS ：S. amoena was extracted with 95％ ethanol to obtain ethanol extract ，and then was respectively extracted with ethyl acetate and n-butanol to obtain corresponding polar parts. Totally 48 mice were randomly divided into normal group （8 mice）and modeling group （40 mice）. Normal group was given constant volume of olive oil intraperitoneally ， 3 times a day ，for consecutive 6 weeks. Model group was given 30％CCl4-olive oil solution intraperitoneally to induce liver injury model，with initial dose of 5 mL/kg after each 3 mL/kg，3 times a days ，for 6 consecutive weeks. After modeling ，the mice were randomly divided into model group （normal saline ），sylibin group （positive control ，20 mg/kg），S. amoena ethanol extract group （100 mg/kg），S. amoena ethyl acetate group （100 mg/kg），and S. amoena n-butanol group （100 mg/kg），with 8 mice in each group. After they were given relevant medicine intragastrically once a day ，for consecutive 6 weeks. The general information during experiment of mice was observed. 1 h after last medication ，the serum contents of TC ，TG，ALT and AST were determined by Enzyme-labelled meter . After HE staining ，the pathological changes of liver tissue were observed and Ishak score was performed. RESULTS：In normal group ，mice had normal activity ，thick and glossy hair ，and the body weight was increased. The liver tissue had no obvious pathological changes. The model group had sparse hair ，and they were emaciated and listlessness ；and body weight （before medication ，1，2 week after medication ）was significantly lower than normal group （P＜0.05 or P＜0.01）. Compared with normal g roup，the contents of TC ，TG，ALT and AST in serum were increased significantly （P＜0.01 or P＜0.05）. The structure of hepatic lobule was severely damaged and had more inflammatory cell infiltration ；the arrangement of hepatic cord FF117（-022）] was disordered and the Ishak score was significantly increased qq.com （P＜0.001）. Compared with model group ，above symptom and liver injury of mice in different administration groups wer improved to different extents. The serum contents of TC ，ALT and AST in silybin group and S. amoena ethyl acetate group ，serum contents of TG in administration groups as well as Ishak scores of liver tissue were decreased significantly in silybin group ，S. amoena ethanol extract group and S. amoena ethyl acetate group （P＜0.05 or P＜0.001）. CONCLUSIONS ：S. amoena ethanol extract and its different solvent parts can protect liver tissue of CCl4-induced liver injury model mice ，and active part is the ethyl acetate part of S. amoena .

OBJECTIVE To study the improvement effects of different polar parts fro m total f lavonoids of Scutellaria amoena on non-alcoholic fatty liver disease （NAFLD）model rats. METHODS The total flavonoids of S. amoena （SAF）were extracted by reflux extraction with ethanol ，suspended with water ，and then extracted with ethyl acetate and n-butanol in order to obtain the extraction parts of SAF （recorded as SAFA and SAFB respectively ）. Thirty-six rats were randomly divided into normal group （n＝ 6）and modeling group （n＝30）. Modeling group was given high-lipid diet to induce NAFLD model. After modeling ，modeling group was randomly divided into model group （normal saline ），fenofibrate group （positive control ，20 mg/kg），SAF group （300 mg/kg），SAFA group （300 mg/kg）and SAFB group （300 mg/kg）；they were given relevant intragastical administration ，once a day，for consecutive 6 weeks. After last administration ，the liver index was calculated ；the levels of total cholesterol （TC）， triacylglycerol（TG），aspartate transaminase （AST），alanine transaminase （ALT），high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C） in serum ，the levels of superoxide dismutase （SOD），glutathione peroxidase （GSH-Px），malondialdehyde（MDA），interleukin-1β（IL-1β），IL-6 and tumor necrosis factor-α（TNF-α）in liver tissue were detected；the pathomorphological changes of liver tissue were observed. RESULTS Compared with normal group ，the liver index ， the levels of TC ，TG，AST，ALT，LDL-C，MDA，IL-1β， IL-6 and TNF-α in serum/liver tissue of model group were all increased significantly （P＜0.05）， while the levels of HDL-C，SOD and GSH-Px were all decreased significantly （P＜0.05）. Compared with model group ，except there was no statistical significance in the serum levels of HDL-C and ALT in SAFA group （P＞0.05），above indexes in serum/liver tissue of rats in groups of polar parts from total flavonoids of S. amoena were significantly improved （P＜0.05）；inflammatory cell infiltration and fatty vacuoles in liver tissue were significantly improved. Compared with SAF group and SAFA group ，the levels of TC，TG，AST，MDA，IL-6 and TNF-α were decreased significantly in SAFB group（P＜0.05），while the level of SOD was increased significantly （P＜0.05）；pathomorphological changes of liver tissue were improved more significantly. CONCLUSIONS Each polar part from total flavonoids of S. amoena can improve NAFLD by regulating oxidative stress and inhibiting the secretion of inflammatory factors. The n-butanol polar part has more obvious effect .

Objective To prepare and characterize the mouse polyclonal antibody against the dense granule protein 24 (GRA24) of Toxoplasma gondii, and explore its preliminary applications. Methods The GRA24 coding sequences of different T. gondii strains were aligned using the MEGA-X software, and the dominant peptide of the GRA24 protein was analyzed with the Protean software. The base sequence encoding this peptide was amplified using PCR assay and ligated into the pET-28a vector, and the generated GRA24 truncated protein was transformed into Escherichia coli BL21. After induction by isopropyl-beta-D-thiogalactopyranoside (IPTG), the expression and purification of the recombinant GRA24 protein was analyzed using sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE). BALB/c mice were immunized by subcutaneous injection with the purified recombinant GRA24 truncated protein to generate the polyclonal antibody, and the titer of the polyclonal antibody was measured using enzyme linked immunosorbent assay (ELISA). The specificity of the polyclonal antibody was tested using Western blotting, and the intracellular localization of the polyclonal antibody was investigated using immunofluorescence assay (IFA). Results SDS-PAGE showed successful construction of the recombinant expression plasmid, and Coomassie brilliant blue staining showed the generation of the high-purity recombinant GRA24 truncated protein. ELISA measured that the titer of the polyclonal antibody against the GRA24 truncated protein was higher than 1:208 400, and Western blotting showed that the polyclonal antibody was effective to recognize the endogenous GRA24 proteins of different T. gondii strains and specifically recognize the recombinant GRA24 truncated protein. Indirect IFA showed that the GRA24 protein secreted 16 hour following T. gondii invasion in host cells. Conclusions The polyclonal antibody against the T. gondii GRA24 protein has been successfully prepared, which has a widespread applicability, high titers and a high specificity. This polyclonal antibody is available for Western blotting and IFA, which provides the basis for investigating the function of the GRA24 protein.