Objective To observe the mRNA and protein expression of wingless-type MMTV integration site family member 5a (Wnt5a), glycogen synthase kinase 3 (GSK3), and β-catenin, as well as the muscle fibers and adipose tissue presented in pathological staining in the gastrocnemius muscle of white rabbits with blunt gastrocnemius contusion injuries, and provide a basis for revealing the repair mechanism of the pressing and kneading method in treating skeletal muscle injury. Methods Forty-two healthy male and female New Zealand white rabbits were selected. They were randomly divided into blank group, model 3-day group, model 7-day group, model 14-day group, press-and-knead 3-day group, press-and-knead 7-day group, and press-and-knead 14-day group, by using a random number table method, with 6 rabbits in each group. Samples of the model groups and the press-and-knead groups were taken on the 4th, 8th and 15th days after operation. The mRNA and protein expression of Wnt5a, GSK3, and β-catenin were detected by quantitative polymerase chain reaction and Western blot; the muscle tissue myofibers and adipose tissue were observed by hematoxylin and eosin (HE) staining and oil red O staining. Results The HE staining results showed that significant fibrous tissue proliferation and inflammatory cell infiltration occurred in the model 7-day group; in the model 14-day group, some muscle fibers were degenerated, necrotic, and regenerated, accompanied by fibrous tissue proliferation, slight inflammatory cell infiltration, and slight calcification; in the press-and-knead groups, obvious muscle fiber degeneration, necrosis, and regeneration, and inflammatory cell infiltration were observed, accompanied by significant fibrous tissue proliferation. The oil red O staining results showed that adipocyte deposition was visible in the model groups, which was the heaviest in the model 7-day group; in the press-and-knead groups, muscle fibers and sequences were not significantly damaged, and a small amount of adipocyte infiltration was visible in the interstitial space. There were statistically significant differences in the mRNA expression and protein expression of Wnt5a, GSK3, and β-catenin in the gastrocnemius among groups (P<0.001). Conclusions The histopathological changes of gastrocnemius muscle injury recover gradually over time, and the pressing and kneading method stimulates the mRNA expression activities of Wnt5a, GSK3, and β-catenin, which may slow down the degradation of β-catenin protein by the scaffolding protein complex (of which GSK3 is an important component), so that the protein level of β-catenin is maintained in the stable range at all times. This leads to a reduction of fatty degeneration in the gastrocnemius muscle after the intervention of pressing and kneading method, and promotes the functional repair of the injured skeletal muscle.
【摘要】 目的 觀察循膀胱經彈撥法干預亞健康狀態方案的療效及優勢，并規范其技術標準，為臨床提供安全有效的治療技術范例。 方法 2009年3月-2010年8月，采用多中心、分層區組隨機、平行對照的臨床研究方法，選擇亞健康狀態受試者360例，隨機分為傳統組、彈撥組、復合組，每組120例。觀察受試者干預前后的臨床癥狀積分、血沉、血漿乳酸、全血黏度（切變率分別為1、5、30、200/s）的變化，及有效性和安全性。 結果 共305例完成試驗，其中傳統組97例，彈撥組102例，復合組106例。彈撥組、復合組的總體療效與傳統組比較差異有統計學意義（Plt;0.05），彈撥組與復合組比較差異無統計學意義（Plt;0.05）。干預后彈撥組、復合組的臨床癥狀積分、血沉、血漿乳酸、全血黏度與傳統組比較差異有統計學意義（Plt;0.05）；彈撥組與復合組臨床癥狀積分、血液乳酸全血黏度（切變率分別為1、200/s）差異無統計學意義（Pgt;0.05），血沉、全血黏度（切變率分別為5、30/s）差異有統計學意義（Plt;0.05）。各組各指標干預后前后自身比較差異均有統計學意義（Plt;0.05）。 結論 推拿能有效地改善亞健康狀態，復合組療效最佳，彈撥組其次，傳統組較差。循膀胱經彈撥法對亞健康狀態的干預有效性較傳統推拿更好。【Abstract】 Objective To observe the efficacy and advantages of flicking and poking along bladder channels in intervening sub-health state and regulate the technical standards for clinical treatment techniques, in order to provide safe and effective treatment criteria. Methods Multi-centered, stratified randomized and parallel controlled clinical research methods were adopted in this study. From March 2009 to August 2010, 360 subjects with sub-health status were randomly divided into traditional group, flicking and poking group, and combining group with 120 cases in each group. The changes of clinical symptoms, erythrocyte sedimentation rate, plasma lactate, and whole blood viscosity (shear rates of 1, 5, 30, 200/s) as well as the efficacy and safety were observed. Results A total of 305 patients completed the trial, including 97 in the traditional group, 102 in the flicking and poking group, and 106 in the combining group. The general efficacy for the flicking and poking group and the combining group was significantly different from that for the traditional group (Plt;0.05), while there was no significant difference between the flicking and poking group and the combining group. After the intervention, the clinical symptom score, erythrocyte sedimentation rate, plasma lactate, and whole blood viscosity of the flicking and poking group and the combining group were significantly different from those of the traditional group (Plt;0.05). The clinical symptom score, plasma lactate and whole blood viscosity (shear rates of 1, 200/s) of the flicking and poking group and the combining group were not significantly different (Pgt;0.05), while erythrocyte sedimentation rate, and whole blood viscosity (shear rates of 5, 30/s) between those two groups were significantly different (Plt;0.05). All the above mentioned indexes before and after intervention in each group were statistically different (Plt;0.05). Conclusion Massage can improve the sub-health state, and the combining method has the best efficacy followed by flicking and poking, while traditional method has a poor efficacy. Intervening sub-health state with flicking and poking along bladder channels has a better efficacy than the traditional method of massage.
Objective To observe the effect of “Luo’s Roujin Technique” on the inflammatory response and joint capsule fibrosis in white rabbits with scapulohumeral periarthritis model. Methods Thirty healthy male New Zealand white rabbits were randomly divided into a control group, a model group, and a treatment group, with 10 rabbits in each group. Scapulohumeral periarthritis models were established in the model group and the treatment group, while the control group received identical restraint procedures at the same timepoints. Six rabbits in the model group and seven in the treatment group were successfully modeled. The subsequent experiment included all six successfully modeled rabbits from the model group, along with six rabbits randomly selected from each of the control and treatment groups. On the second day after successful modeling, blood samples were collected from the auricular marginal vein in all three groups. After blood collection, the treatment group began massage therapy for 21 consecutive days, while the other two groups underwent the same restraint procedure simultaneously. On Day 22, all the three groups were euthanized after blood collection from the auricular marginal vein, and the synovial tissue of the affected shoulder joint was completely collected. Hematoxylin-eosin staining was used to examine the histopathological features of the synovial tissue. Enzyme-linked immunosorbent assay was employed to measure the concentrations of interleukin (IL)-1β, IL-6, IL-17, and tumor necrosis factor-α (TNF-α). Western blot and reverse transcription polymerase chain reaction were used to assess the protein and mRNA expression levels of vascular endothelial growth factor (VEGF), connective tissue growth factor (CTGF), transforming growth factor-β1 (TGF-β1), and Smad3. Results After treatment, the control group showed no significant inflammatory cell infiltration or fibrous tissue proliferation in the synovial tissue. The model group exhibited synovial cell hyperplasia in the lining layer and inflammatory cell infiltration in the sublining layer. The treatment group displayed mild inflammatory cell infiltration in the sublining layer. Compared with the control group, the model group showed significantly increased concentrations of IL-1β, IL-6, IL-17, and TNF-α in both serum and synovial homogenate, as well as elevated protein and mRNA expression of VEGF, CTGF, TGF-β1, and Smad3 in synovial tissue (P<0.05). Compared with the model group, the treatment group exhibited significantly lower serum levels of IL-1β, IL-6, and TNF-α, as well as reduced synovial homogenate levels of IL-1β, IL-6, IL-17, and TNF-α (P<0.05); furthermore, protein expression of VEGF, CTGF, TGF-β1, and Smad3 and mRNA expression of VEGF and CTGF in synovial tissue were significantly decreased in the treatment group (P<0.05). Conclusions “Luo’s Roujin Technique” can significantly alleviate local inflammatory infiltration in the synovial tissue of rabbits with scapulohumeral periarthritis, and reduce the levels of IL-1β, IL-6, IL-17, and TNF-α in both serum and synovial tissue. The underlying mechanism may involve suppression of VEGF, CTGF, TGF-β1, and Smad3 expression, leading to attenuated inflammatory responses and inhibition of fibroblast-to-myofibroblast transition. Thereby, it mitigates fibrotic changes in the shoulder joint capsule, exerting anti-inflammatory and analgesic effects and improving joint mobility.
Objective
To investigate the clinical characteristics of allergic and non-allergic asthma in Chinese adult asthmatic patients.
Methods
Consecutive treatment-naive adult outpatients with asthma were retrospectively analyzed in West China Hospital, Sichuan University from October 2014 to June 2016. The patients were classified into a non-allergic asthma (NAAS) group and an allergic asthma (AAS) group by skin prick test or antigen-specific IgE test. The differences between allergic and non-allergic asthma were compared in respect of gender, age, asthma control test (ACT) score, lung function, fractional exhaled nitric oxide (FeNO) level, body mass index (BMI), disease severity,etc.
Results
A total of 131 patients were enrolled in which 72 cases (54.96%) were allergic asthmatics and 59 cases (45.04%) were non-allergic asthmatics. The level of FeNO was statistically different (t=–2.762,P=0.007) between the NAAS group and the AAS group [(51.1±32.6)ppbvs. (69.1±41.7)ppb]. Seventeen cases of the NAAS group and 48 cases of the AAS group were complicated with rhinitis with statistically significant difference (χ2
=19.396,P=0.000). Airway limitation reversibility test showed that there were 37 cases in AAS and 20 cases in NAAS with no airway obstruction (NAO), 26 cases in AAS and 22 cases in NAAS with reversible airflow obstruction (RAO), 9 cases in AAS and 17 cases in NAAS with irreversible airflow obstruction (IAO), respectively, with statistically significant difference between two groups (Z=–2.461,P=0.014). There were 20 cases (33.9%) in NAAS and 37 cases (51.4%) in AAS with mild intermittent or persistent asthma, 18 cases (30.5%) in NAAS and 19 cases (26.4%) in AAS with moderate persistent asthma, 21 cases (35.6%) in NAAS and 16 (22.2%) in AAS with severe persistent asthma, respectively, with statistically significant difference (Z=–2.115,P=–0.034). The age, ACT score, FEV1%pred, and BMI had no statistical difference between two groups (allP>0.05).
Conclusion
Compared with allergic asthma, non-allergic asthma has less rhinitis, lower FeNO levels and higher prevalence of irreversible airflow obstruction.
【Abstract】ObjectiveTo study the effect of transfection with antisense DNMT3b gene eukaryotic expression vector on the expression of DNMT3b gene in human cholangiocarcinoma cell line QBC-939. MethodsThe constructed antisense DNMT3b gene eukaryotic expression vector was transfected into the human cholangiocarcinoma cell line QBC-939 by using lipofectamine transfection reagents, and positive cell clones were obtained by using G418 selection after transfection. Whether the constructed recombinant vector was transfected into QBC-939 cells successfully was confirmed by amplifying the exogenous neoR gene with PCR method. The expression of DNMT3b gene mRNA and protein were detected by semi-quantitative RT-PCR and FCM methods respectively. ResultsFollowing the transfection of antisense DNMT3b gene eukaryotic expression vector, the mRNA level of DNMT3b gene in QBC-939 cells of human cholangiocarcinoma decreased from 0.956±0.053 to 0.209±0.023, and the protein level of DNMT3b gene also decreased from (75.38±3.22)% to (29.87±3.46)%. There were very significant differences on the expression levels of DNMT3b gene between non-tranfections group and the antisense DNMT3b gene eukaryotic expression vector transfection group (P＜0.01). ConclusionTransfection with antisense DNMT3b gene eukaryotic expression vector significantly reduces the expression level of DNMT3b gene in human cholangiocarcinoma cell line QBC-939, and this study may provide a valid tool and method to investigate the function of DNMT3b gene and its role in cholangiocarcinoma.
Objective To compare the effect of decimeter wave with sodium hyaluronate product (SHP) on preventing and treating peritendinous adhesion and promoting tendon heal ing. Methods Totally 96 healthy male white 6-month-old Leghorn chickens weighing (2.24 ± 0.07) kg were randomized into group A ( decimeter wave therapy group, n=32),in which decimeter wave therapy was appl ied 1 to 21 days after operation at a frequency of 915 MHz, a power of 8 W, radiation distance of 10 cm, for 10 minutes once per day; group B (SHP group, n=32), in which 5 mL and 1.2% SHP was appl ied; and group C (control group, n=32), in which injury received no treatment. The III and IV toes of left feet of all chickens were made into tendon injury model. The general condition of animal was observed after operation; gross and histological observations were made 7, 10, 14, 18, 21 and 28 days after operation, and the biomechanical analysis was done 14 and 28 days after operation. Results Operative incision healed well, no infection and death occurred. Peritendinous adhesions in groups A, B were looser, and tendon heal ing was better than that of group C 14 and 28 days after operation. More fibroblasts with active metabol ism and more collagen formation in groups A, B than that in group C. The Pmax of group A was better than that of group B 14 and 28 days after operation (P lt; 0.05); the δmax of group A was better than that of group B 18 and 21 days after operation (P lt; 0.05), and the W0 of group A was better than that of group B 18, 21 and 28 days after operation (P lt; 0.05). There was no significant difference between group A and group B at the other time points. Conclusion Topical decimeter wave therapy and appl ication of SHP after flexor tendon repair can promote intrinsic heal ing, meanwhile they can prevent the adhesion of tendon and reduce extrinsic heal ing. Decimeter wave therapy can improve the qual ities of tendon’s wound heal ing.
