BACKGROUND:Developed by the esteemed Chinese medicine master Wei Guikang,Tongan Decoction has proven highly effective in treating knee osteoarthritis.However,the mechanism of action is yet unclear.OBJECTIVE:To elucidate how Tongan Decoction modulates synovial macrophage polarization as a therapeutic strategy for knee osteoarthritis in a rat model.METHODS:(1)We employed high-throughput microRNA sequencing and polymerase chain reaction to analyze the differentially expressed miRNA in synovial macrophages of normal and knee osteoarthritis rats.Predicted target genes of miR-27a were identified using bioinformatics databases,with subsequent validation through luciferase assays.A total of 68 Sprague-Dawley rats were randomly divided into normal control group(n=16),model group(n=16),miR-27a overexpression group(n=12),Tongan Decoction group(n=12),and Tongan Decoction+miR-27a inhibition group(n=12).The miR-27a overexpression group and the Tongan Decoction+miR-27a inhibition group were injected with miR-27a mimic and miR-27a inhibitor in the right knee joint cavity,respectively,once daily for 5 continuous days.On the 15th day after modeling,the Tongan Decoction group and the Tongan Decoction+miR-27a inhibition group were given Tongan Decoction by gastric lavage,and the other three groups were given saline by gastric lavage,once daily for 14 continuous days.After administration,behavioral tests,X-rays,hematoxylin-eosin staining of synovial and cartilage tissues of the knee joint and immunofluorescence staining of synovial tissues of the knee joint,RT-PCR,and Western blot were performed.RESULTS AND CONCLUSION:High-throughput sequencing of miRNA showed low expression of miR-27a in synovial tissues of rats with knee osteoarthritis.The target gene of miR-27a was nuclear factor κB,and luciferase assay showed that the two could bind to each other.Behavioral assays showed that miR-27a overexpression or Tongan Decoction alleviated joint dysfunction in rats with knee osteoarthritis,and miR-27a inhibition antagonized the effect of Tongan Decoction.X-ray films and hematoxylin-eosin staining showed that miR-27a overexpression or Tongan Decoction reduced the degree of knee osteoarthritis and miR-27a inhibition weakened the therapeutic effect of Tongan Decoction.The results of RT-PCR and western blot assay showed that compared with the normal control group,the expression of interleukin 10 was reduced in the model group(P＜0.05),and the expression of matrix metalloproteinase 13,interleukin 1β,and nuclear factor κB was elevated in the model group(P＜0.05).miR-27a overexpression or Tongan Decoction could differently reverse the changes in the above-mentioned indexes,while miR-27a inhibition weakened the effect of Tongan Decoction.Immunofluorescence staining results showed that CD86 protein expression in the model group was higher than that in the normal control group(P＜0.05),and CD206 protein expression was lower than that in the normal control group(P＜0.05);miR-27a overexpression group and Tongan Decoction had lower CD86 protein expression than that in the model group(P＜0.05),and higher CD206 protein expression than that in the model group(P＜0.05);in the Tongan Decoction+miR-27a inhibition group,CD86 protein expression was higher(P＜0.05)and CD206 protein expression was lower than that in the Tongan Decoction group(P＜0.05).Tongan Decoction mitigates knee osteoarthritis by upregulating miR-27a expression and suppressing nuclear factor κB activity,which improves knee joint function and further treats knee osteoarthritis.

BACKGROUND:Developed by the esteemed Chinese medicine master Wei Guikang,Tongan Decoction has proven highly effective in treating knee osteoarthritis.However,the mechanism of action is yet unclear.OBJECTIVE:To elucidate how Tongan Decoction modulates synovial macrophage polarization as a therapeutic strategy for knee osteoarthritis in a rat model.METHODS:(1)We employed high-throughput microRNA sequencing and polymerase chain reaction to analyze the differentially expressed miRNA in synovial macrophages of normal and knee osteoarthritis rats.Predicted target genes of miR-27a were identified using bioinformatics databases,with subsequent validation through luciferase assays.A total of 68 Sprague-Dawley rats were randomly divided into normal control group(n=16),model group(n=16),miR-27a overexpression group(n=12),Tongan Decoction group(n=12),and Tongan Decoction+miR-27a inhibition group(n=12).The miR-27a overexpression group and the Tongan Decoction+miR-27a inhibition group were injected with miR-27a mimic and miR-27a inhibitor in the right knee joint cavity,respectively,once daily for 5 continuous days.On the 15th day after modeling,the Tongan Decoction group and the Tongan Decoction+miR-27a inhibition group were given Tongan Decoction by gastric lavage,and the other three groups were given saline by gastric lavage,once daily for 14 continuous days.After administration,behavioral tests,X-rays,hematoxylin-eosin staining of synovial and cartilage tissues of the knee joint and immunofluorescence staining of synovial tissues of the knee joint,RT-PCR,and Western blot were performed.RESULTS AND CONCLUSION:High-throughput sequencing of miRNA showed low expression of miR-27a in synovial tissues of rats with knee osteoarthritis.The target gene of miR-27a was nuclear factor κB,and luciferase assay showed that the two could bind to each other.Behavioral assays showed that miR-27a overexpression or Tongan Decoction alleviated joint dysfunction in rats with knee osteoarthritis,and miR-27a inhibition antagonized the effect of Tongan Decoction.X-ray films and hematoxylin-eosin staining showed that miR-27a overexpression or Tongan Decoction reduced the degree of knee osteoarthritis and miR-27a inhibition weakened the therapeutic effect of Tongan Decoction.The results of RT-PCR and western blot assay showed that compared with the normal control group,the expression of interleukin 10 was reduced in the model group(P＜0.05),and the expression of matrix metalloproteinase 13,interleukin 1β,and nuclear factor κB was elevated in the model group(P＜0.05).miR-27a overexpression or Tongan Decoction could differently reverse the changes in the above-mentioned indexes,while miR-27a inhibition weakened the effect of Tongan Decoction.Immunofluorescence staining results showed that CD86 protein expression in the model group was higher than that in the normal control group(P＜0.05),and CD206 protein expression was lower than that in the normal control group(P＜0.05);miR-27a overexpression group and Tongan Decoction had lower CD86 protein expression than that in the model group(P＜0.05),and higher CD206 protein expression than that in the model group(P＜0.05);in the Tongan Decoction+miR-27a inhibition group,CD86 protein expression was higher(P＜0.05)and CD206 protein expression was lower than that in the Tongan Decoction group(P＜0.05).Tongan Decoction mitigates knee osteoarthritis by upregulating miR-27a expression and suppressing nuclear factor κB activity,which improves knee joint function and further treats knee osteoarthritis.

Objective To investigate the clinical effect of different magnetic stimulation pelvic floor modes in the treatment of perimenopausal myofascial pelvic pain syndrome(MPSS).Methods A total of 60 perimenopausal women who were clinically diagnosed with MPSS in the hospital from May 2022 to May 2023 were selected as the research objects.They were divided into groups A,B and C by random number table method,with 20 cases in each group.All patients in the three groups were treated with pelvic floor myofascial manual release.Group A was given pelvic floor magnetic stimulation(alternating 10 Hz and 50 Hz),group B was given sacral nerve root magnetic stimulation(50 Hz),and group C was given pelvic floor magnetic stimu-lation combined with sacral nerve root magnetic stimulation at the same time.The three groups were treated twice a week for eight weeks.Visual analogue scale(VAS)was used to evaluate the degree of pelvic floor myofascial tenderness before and after treatment,and Glazer pelvic floor surface electromyography was used to evaluate pelvic floor muscle function.Results Compared with before treatment,the VAS scores of subjec-tive pain and pelvic floor myofascial tenderness in the three groups were decreased after treatment,and the differences were statistically significant(P<0.05).Compared with group A and group B,the VAS score of subjective pain and the VAS score of pelvic floor myofascial tenderness in group C were significantly decreased after treatment,and the differences were statistically significant(P<0.05).Compared with before treatment,the average amplitude and coefficient of variation(CV)of pre-rest potential and post-rest potential in the three groups were decreased after treatment,and the differences were statistically significant(P<0.05).Compared with before treatment,only the maximum amplitude of rapid contraction,the average amplitude of 10 s sustained contraction and 60 s sustained contraction and CV in group C were improved,and the differ-ences were statistically significant(P<0.05).Compared with group A and group B,the average amplitude and CV of pre-resting potential and post-resting potential in group C were decreased after treatment,the maxi-mum amplitude of rapid contraction and the average amplitude and CV of 10 s continuous contraction and 60 s persistent contraction were improved,and the differences were statistically significant(P<0.05).Conclusion Dif-ferent magnetic stimulation pelvic floor modes can effectively relieve pain and improve pelvic floor muscle strength in the treatment of perimenopausal MPSS,and the effect of pelvic floor magnetic stimulation com-bined with sacral nerve root magnetic stimulation is the best.

Objective:To investigate the effect of recombinant insulin-like growth factor-1 (IGF-1) on ovarian development in MEX3C knockout mice. Methods:Mice were randomly divided into four groups (6 mice in each group): wild type group, homozygous group, homozygous+IGF-1 treatment group, and treatment control group. The weight and wet ovarian weight of mice in each group was compared before and after IGF-1 injection. The serum levels of estradiol was determined using enzyme linked immunosorbent assay (ELISA). The morphological structure of the ovary was observed by HE staining. The expression of MEX3C, IGF-1 receptor (IGF-1R), and the phosphorylation of AKT (Ser473)(p-AKT) were detected by immunohistochemistry and Western blotting.Results:Before IGF-1 injection, the body weight of homozygote group [(6.33±0.31) g] was significantly lighter than that in wild type group [(15.20±0.35) g, P<0.001]. Serum estradiol level was significantly higher in homozygous+IGF-1 treatment group [(17.245 3±0.073 1) ng/L] than in treatment control group [(8.124 8±0.847 7) ng/L, P=0.001]. HE staining results showed that compared with the treatment control group, the primitive follicles (19.83±2.94) and primary follicles (15.50±2.69) were increased in homozygous+IGF-1 treatment group, while the atretic follicles (7.17±1.42) were decreased (all P<0.05). Western blotting results further confirmed that compared with the treatment control group, IGF-1R and p-AKT protein expressions were significantly decreased in homozygous+IGF-1 treatment group ( P=0.014 1, P=0.002 5). Conclusion:IGF-1 promotes ovarian follicular development in MEX3C knockout mice. The mechanism is related to the up-regulation of IGF-1R and P-AKT protein expression. IGF-1 can increase serum estrogen and promote follicular development.

Objective:To investigate the effect of recombinant insulin-like growth factor-1 (IGF-1) on ovarian development in MEX3C knockout mice. Methods:Mice were randomly divided into four groups (6 mice in each group): wild type group, homozygous group, homozygous+IGF-1 treatment group, and treatment control group. The weight and wet ovarian weight of mice in each group was compared before and after IGF-1 injection. The serum levels of estradiol was determined using enzyme linked immunosorbent assay (ELISA). The morphological structure of the ovary was observed by HE staining. The expression of MEX3C, IGF-1 receptor (IGF-1R), and the phosphorylation of AKT (Ser473)(p-AKT) were detected by immunohistochemistry and Western blotting.Results:Before IGF-1 injection, the body weight of homozygote group [(6.33±0.31) g] was significantly lighter than that in wild type group [(15.20±0.35) g, P<0.001]. Serum estradiol level was significantly higher in homozygous+IGF-1 treatment group [(17.245 3±0.073 1) ng/L] than in treatment control group [(8.124 8±0.847 7) ng/L, P=0.001]. HE staining results showed that compared with the treatment control group, the primitive follicles (19.83±2.94) and primary follicles (15.50±2.69) were increased in homozygous+IGF-1 treatment group, while the atretic follicles (7.17±1.42) were decreased (all P<0.05). Western blotting results further confirmed that compared with the treatment control group, IGF-1R and p-AKT protein expressions were significantly decreased in homozygous+IGF-1 treatment group ( P=0.014 1, P=0.002 5). Conclusion:IGF-1 promotes ovarian follicular development in MEX3C knockout mice. The mechanism is related to the up-regulation of IGF-1R and P-AKT protein expression. IGF-1 can increase serum estrogen and promote follicular development.

Objective To observe the influence of ultra-shortwave (USW) irradiation on infarct volume and Ca2+-ATPase (SPCA) secretion after brain ischemia and reperfusion.Methods Eighty Sprague-Dawley rats were randomly divided into a sham operation group (n=8),a model group (n=36) and a USW group (n=36).The animal model of middle cerebral artery ischemia and reperfusion (MCAO/R) was established using the suture method in the rats of the model and USW groups,while the sham operation group was given the same operation but without inserting the thread plug.One day,3 days and 7 days after the intervention,12 rats were sacrificed and the infarct volumes and SPCA1 protein expression were measured using 2,3,5-triphenyltetrazolium chloride staining and western blotting.Results No white infarcted tissue was found in the sham operation group.In the model and USW groups the volume of infarcted tissue decreased with time.Significantly less infarcted volume was observed in the USW group compared to the model group at each time point.The SPCA1 levels in the brain tissue were lower than in the sham operation group after one and 3 days of USW treatment,but they were significantly lower in the model group as well.As time went by,the average SPCA1 level increased significantly in the model and USW groups.A slightly higher SPCA1 level was observed in the USW group compared to the model group after one day of treatment,but with no significance.However,significant differences were found between them after 3 and 7 days of intervention.Conclusion Ultra-shortwave irradiation can protect against MCAO/R injury by decreasing the infarcted volume,which may be related to down-regulation of SPCA1,minimizing nerve cell apoptosis and promoting neural functional recovery,at least in rats.

To develop a specific, rapid and convenient method based on molecular motor biosensor to detect food-borne rotavirus. A specific probe was encompassed the conservative region of rotavirus's VP7 segment, and a molecular motor detect device was constructed by connecting probes to F0F1-ATPase molecular motor through biotin-streptavidin system. This biosensor's sensitivity was 0.005 ng/mL for rotavirus RNA. Extracted virus RNA was conjugated with the biosensor separately, at the same time ATP was synthesized. By comparing fluorescence intensity, we can detect rotavirus RNA in samples. This method possessed specificity for rotavirus, without any cross-reaction with Hepatitis A virus and noroviris, and it could be accomplished within 1 h. We detected 15 samples using this method and the results were compared with RT-PCR results. This method is sensitive and specific for rotavirus, and it can be used to detect food-borne rotavirus.
Biosensing Techniques ; methods ; DNA, Viral ; analysis ; genetics ; Food Microbiology ; methods ; Rotavirus ; genetics ; isolation & purification ; Sensitivity and Specificity