Objective To investigate the significance of serum interleukin-1β,interleukin-2,interleukin-6,TNF-α,erythrocyte sedimentation rate(ESR),c-reactive protein(CRP),IgA,IgG and IgM in patients with ankylosing spondylitis(AS),and to analyze their relationship with the Bath ankylosing spondylitis disease activity index (BASDAI). Methods Participants of this study included 45 AS patients(patient group)and 30 healthy subjects (control group).The patient group was further divided into an active subgroup and an inactive subgroup based on the disease activity assessed by using BASDAI.Then serum levels of IL-1β,IL-2,IL-6,TNF-α,ESR,CRP,IgA,IgC and IgM were tested in all the subjects,and the values were compared between different groups.In addition,the relationship between various parameters and those with BASDAI were also evaluated. Results The serum levels of ESR,CRP,IgA,lgM,IgG,IL-1β,IL-2,IL-6 and TNF-α in patients with AS were significantly higher than those in the healthy controls(P<0.05).The serum level of CRP,IL-2 and IL-6 were significantly different between the active and inactive subgroups(P<0.05).There was a positive correlation between CRP and IL-6,IL-6 and IL-2,as well as IgA and IgG levels.The IL-2,IL-6 and CRP levels were positive correlated with BASDAI.Conclusion It was suggested that the serum levels of IL-2,IL-6 and CRP can be used to assess the disease activity in patients with AS.

Objective Take frizzled class receptor 4(FZD4) as an example for exploring whether the transmembrane receptor protein is a suitable marker for wound age estimation. Methods A total of 78 Sprague-Dawley rats were divided randomly into the control group and the contusion groups of 4h, 8h, 12h, 16h, 20h, 24h, 28h, 32h, 36h, 40h, 44h, and 48h after injury (n=6). Using a drop-ball technique, a contusion was produced in the right lamb of rats. The samples were dissected from injury zones. Then the expression of FZD4 was detected using immunofluorescence, real-time PCR and western blot, respectively. Results FZD4 was located on the membrane of skeletal muscle cells. Compared to the control group, FZD4 mRNA showed a signiifcant increase (over 2-fold) in 8h, 12h, 36h, and 40h after injury by Real-time PCR (P<0.05), and FZD4 protein showed a statistical up-regulation (less than 2-fold) in 8h, 36h, 40h, 44h, and 48h post contusion by western blotting (P<0.05). Conclusion The expression of FZD4 mRNA and protein are both time-dependent during contused skeletal muscle repair. To some degree, FZD4 mRNA was more suitable than corresponding protein for determining wound age.

Objective T o investigate the relation betw een injury tim e and the expression of cytochrom e c oxidase subunitⅥc (COX6C) m R N A in skeletal m uscle of rat after contusion. Methods A total of fifty-four SD rats w ere divided into the control group and the contusion groups (0.5,1,6,12,18, 24, 30, and 36 h after contusion), random ly. T he contusion m odel w as established by free fall drop of gravity ham-m er. A t corresponding tim e point after contusion, the regular histology w as exam ined and expression level of COX6C m R N A w as tested by real-tim e PC R after extraction of total R N A from the tissues. Results T he m ain pathological features of 6 h after injury included edem a and hem orrhage in m yocytes w ith no inflam m atory cells found. A fter 6 hours, the findings included m yocyte degeneration and necro-sis, inflam m atory cells infiltration, and fibrous connective tissue proliferation in the contused zone. T he expression level of COX6C m R N A w as higher than that of the control group w ithin 6 h after contusion. T he expression level w as low er than that of the control group from 6-36 h after contusion. Conclusion T he level of COX6C m R N A expresses in a regular w ay after contusion. It m ay be useful for estim ating w ound age in com bination w ith the results of pathological features.

Objective In order to understand which kind of function genes play an important role for es-timating wound age, the variation of difference genes’ mRNA expression were compared after injury. Methods T he mRNA expression levels of seven candidate genes (ICAM-1, NF-κB, MX2, MT1, MT2, sTnI, and Cox6c) were analyzed in contused rat skeletal muscle at different time points using real-time fluorescent quantitative PC R (R T-qPC R ). T he rawC t values were normalized relative to that of RPL32 mRNA , and converted to standard C t values. A t each time point after injury, the standard deviations (SD ) of the standard C t values were calculated by SPSS. Results T he expression trends of the seven genes were all found to be related to wound age, but there were lower variation coefficients and greater reliability of sTnI and Cox6c when compared with other genes. Conclusion T he genes encoding struc-tural proteins or proteins that performbasic functions can be suitable for wound age estimation.

Objective T o investigate the relationship betw een the expression of secreted frizzled-related protein 5 (SFR P5) m RNA and the tim e interval after skeletal m uscle injury in rats by real-tim e PC R . Methods A total of ninety SD rats w ere random ly divided into the contusion groups at different tim es including 4 h, 8 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, 48 h after contusion, incision groups at different tim es including 4h and 8h after incision and the control group. T he sam ples w ere taken from the contused zone at different tim e points. T he total RNA w as isolated from the sam ples and re-versely transcribed to analyze the expression levels of SFRP5 m RNA . Results C om pared to the control group, the expression of SFRP5 m RNA in contusion groups w ere dow n-regulated w ithin 48 h after con-tusion and reached the low est level at 20 h, and the expression of SFRP5 m RNA gradually increased from 20 h to 48 h after contusion. T he expression of SFRP5 m RNA in the incised groups w ere signifi-cantly low er than that of the contusion groups at 4 h after injury. A t the tim e of 8 h, the expression levels betw een the contusion and incision groups show ed no statistically significant difference. Conclusion It is suggested that SFRP5 m RNA analysis m ay show regular expression and can be a m arker for estim ation of skeletal m uscle injury age.

Objective To explore the clinical efficacy of arc-edge needle therapy based on Zaodian theory in the treatment of refractory tennis elbow.Methods 64 patients with refractory tennis elbow were collected and randomly divided into a control group and an observation group using a random number table method.Among them,32 cases in the control group were treated with local pain point injection.32 patients in the observation group were treated with arc-edge needle therapy once a week,with two treatments per group.The Number rating scale(NRS)Mayo,elbow joint performance score(MEPS)and Disabilities of the arm,shoulder and hand(DASH)score were recorded and compared between the two groups before treatment and at 1,2,4 and 24 weeks of follow-up after treatment,disease efficacy score(WDES)at 24 weeks of follow-up.Results During the treat-ment process,neither group of patients experienced serious adverse reactions.At each stage after treatment,two groups of NRS MEPS,DASH,and WDES at 24 weeks of follow-up all improved compared to before treatment.After one week of treatment,there was no statistically significant difference in NRS scores between the two groups(P>0.05).At follow-up visits at weeks 2,4,and 24,the NRS scores in the observation group were lower than those in the control group.After 1,2,4,and 24 weeks of follow-up after treatment,the DASH score was lower in the observation group than in the control group,and the MEPS score was higher in the observation group than in the control group,with statistically significant differences(P<0.05).After 24 weeks of treatment,the WDES of the observation group was better than that of the control group,and the difference was statistically significant(P<0.05).Conclusion Local pain point injection and arc-edge Zaodian release can alleviate pain and improve elbow joint function in the early stage of refractory tennis elbow.However,the long-term effect of local pain point injection treatment is not good,and the treatment effect of the observation group gradually improves.

Objective To address the challenges of poor performance and lack of interpretability in existing models,the SHAP algorithm is used to develop an interpretable machine learning model that offers a novel approach to wound age estimation,Methods Based on the previous discovery of the expression of 35 wound age healing-related genes in contused skeletal muscle,the woun age estimaton model was constructed using four algorithms,namly,Multilayer Perceptron(MLP),Random Forest(RF),LightGBM(LGBM),and Support Vector Machine(SVM).The SHAP(Shapley Additive Explanation)algorithm was used to rank the importance of genetic features,eliminate redundant attributes,and optimize the model for accurate wound age estimation.the genetic features of the optimal model were analyzed using SHAP's local interpretation capabilities.Results The best results were obtained using model of MLP(area under the curve(AUC)=0.99)The wound ages were classified into four categories:4～12 h,16～24 h,28～36 h,and 40～48 h,using only 15 gene features.According to SHAP analysis,Fam210a was identified as the most relevant gene.Local analysis revealed that high expression of Fam210a contributed to an increase in the predicted probability of 4 h～12 h,while high expression of Rae1 contributed to an increase in the predicted probability of 16 h～24 h.Additionally,low expression of Tbx18 contributed to an increase in the predicted probability of 28 h～36 h,whereas high expression of Tbx18 contributed to an increase in the predicted probability of 40 h～48 h.Conclusions The combined MLP and SHAP model can be used to predict wound age.Using the SHAP interpreter can better understand the degree of contribution of feature genes to the model prediction,and lay the foundation for further in-depth study of wound age estimation.