BACKGROUND:Lijin manipulation can reduce fibrosis scar hyperplasia and promote skeletal muscle repair.However,improper activation of the Wnt/β-catenin signaling pathway can aggravate the fibrosis of injured skeletal muscle and adversely affect the repair process of skeletal muscle.To study the regulatory effect of Lijin manipulation on the Wnt/β-catenin signaling pathway is conducive to elucidate the related mechanisms of Lijin manipulation in reducing fibrosis scar hyperplasia and promoting skeletal muscle injury repair.OBJECTIVE:To explore the mechanism of Lijin manipulation in promoting the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group with 15 rabbits in each group.Gastrocnemius muscle percussion modeling was performed in both model group and Lijin group.Lijin manipulation was performed in the Lijin group on the 3rd day after modeling,once a day,15 minutes once.Five animals in each group were selected and killed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining and the content of collagen fiber was observed by Masson staining.Western blot was used to detect the protein expression of Wnt3a,β-catenin,GSK3β,p-GSK3β,TCF,type I collagen and type III collagen in gastrocnemius muscle,and RT-PCR was used to detect the mRNA expression of Wnt3a,β-catenin and TCF.The expression of β-catenin was detected by immunofluorescence,and the expression of type I collagen and type III collagen was detected by immunohistochemistry.RESULTS AND CONCLUSION:The results of hematoxylin-eosin staining and Masson staining showed that compared with the model group,inflammatory cell infiltration and collagen fiber amount decreased in the Lijin group(P<0.001),and muscle fibers gradually healed.Western blot results showed that compared with the model group,the protein expression levels of Wnt3a,β-catenin,TCF,type I collagen and type III collagen were significantly decreased in the Lijin group at all observation time points(P<0.05),while the ratio of P-GSK3β/GSK3β was significantly increased in the Lijin group at all observation time points compared with the model group(P<0.05).RT-PCR results showed that compared with the model group,the mRNA expression levels of Wnt3a,β-catenin and TCF were significantly decreased in the Lijin group at all observation time points(P<0.001).Immunofluorescence results showed that compared with the model group,the fluorescence intensity of β-catenin expression in the Lijin group was significantly decreased at each observation time point and gradually became similar to that in the blank group(P<0.001).Immunohistochemical results showed that the expression levels of type I collagen and type III collagen in the Lijin group were significantly lower than those in the model group(P<0.01).To conclude,Lijin manipulation could inhibit the abnormal activation of the Wnt/β-catenin signaling pathway,reduce fibrotic scar hyperplasia,and promote the repair of injured skeletal muscle.

BACKGROUND:Excessive apoptosis in skeletal muscle cells will destroy the dynamic balance of the number of myocytes,leading to pathological injury of skeletal muscle.Lijin manipulation is effective in treating skeletal muscle injury,but whether it can inhibit apoptosis and promote the repair of skeletal muscle injury is unknown.OBJECTIVE:To explore the mechanism by which Lijin manipulation reduces inflammation and apoptosis during the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group(n=15 per group).No intervention was performed in the blank group.Gastrocnemius muscle percussion molding was performed in both the model group and Lijin group.After modeling,the model group was not treated,while the Lijin manipulation(Stroking,kneading,and rubbing)was performed in the Lijin group on the 3rd day,once a day,15 min/time.Sampling in each group was performed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining.The ultrastructure of gastrocnemius muscle was observed by transmission electron microscopy.Apoptosis of gastrocnemius cells was observed by TUNEL staining.The expressions of interleukin-1β,interleukin-6 and interleukin-10 in gastrocnemius muscle were detected by ELISA.The protein expressions of BAX,BCL-2 and Caspase3 in gastrocnemius muscle were detected by western blot.The mRNA expression of BAX and BCL-2 was detected by RT-PCR.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining results showed that compared with the model group,inflammatory cells decreased in number,myocyte amount increased,and muscle tissue gradually healed in the Lijin group at each observation point.(2)The results of transmission electron microscopy showed that compared with the model group,the arrangement of muscle fibers at each observation point in the Lijin group was gradually orderly,mitochondria were gradually complete,Z-line arrangement was gradually regular,and free ribosomes were gathered.(3)TUNEL staining results showed that compared with the model group,apoptosis rate in the Lijin group was gradually decreased at all observation points(P<0.05).(4)ELISA results showed that compared with the model group,the expression of interleukin-1β and interleukin-6 in the Lijin group continued to decrease(P<0.05),while the expression of interleukin-10 increased on the 7th day after modeling,and then showed a downward trend(P<0.05).(5)Western blot results showed that compared with the model group,the expression of BCL-2 protein/BAX protein in the Lijin group was significantly increased at each observational point(P<0.05).The protein expression of Caspase3 decreased significantly(P<0.001),and was gradually similar to that of the blank group.(6)RT-PCR results showed that compared with the model group,the mRNA expression level of BCL-2/BAX in the Lijin group was significantly higher at each observational point(P<0.05).To conclude,Lijin manipulation can inhibit inflammation,reduce apoptosis,and promote the repair of injured skeletal muscle.

BACKGROUND:Lijin manipulation can promote skeletal muscle repair and treat skeletal muscle injury.However,the formation of fibrosis and scar tissue hyperplasia are closely related to the quality of skeletal muscle repair.To study the regulatory effect of Lijin manipulation on the formation of fibrosis and scar tissue hyperplasia is helpful to explain the related mechanism of Lijin manipulation to improve the repair quality of skeletal muscle injury. OBJECTIVE:To explore the mechanism of Lijin manipulation to improve the repair quality of skeletal muscle injury in rabbits,thereby providing a scientific basis for clinical treatment. METHODS:Forty-five healthy adult Japanese large-ear white rabbits were randomly divided into blank group,model group and Lijin group,with 15 rats in each group.Gastrocnemius strike modeling was performed in both model group and Lijin group.The Lijin group began to intervene with tendon manipulation on the 3rd day after modeling,once a day,and 15 minutes at a time.Five animals in each group were killed on the 7th,14th and 21st days after modeling.The morphology and inflammatory cell count of gastrocnemius were observed by hematoxylin-eosin staining,the collagen fiber amount was observed by Masson staining,the expression of interleukin-6 and interleukin-10 in gastrocnemius was detected by ELISA.The protein and mRNA expressions of paired cassette gene 7,myogenic differentiation factor,myoblastogenin,alpha-actin,transforming growth factor beta 1,and type Ⅰ collagen were detected by western blot and RT-PCR,respectively,and the expression of type Ⅰ collagen protein was detected by immunohistochemistry. RESULTS AND CONCLUSION:Hematoxylin-eosin staining and Masson staining showed that compared with the model group,inflammatory cell infiltration and collagen fiber content decreased in the Lijin group(P<0.01),and the muscle fibers gradually healed.ELISA results showed that compared with the model group,the expression of interleukin-6 in the Lijin group continued to decrease(P<0.05),and the expression of interleukin-10 increased on the 7th day after modeling(P<0.05)and then showed a decreasing trend(P<0.05).Western blot and RT-PCR results showed that compared with the model group,the protein and mRNA expressions of paired cassette gene 7,myogenic differentiation factor,myoblastogenin in the Lijin group were significantly increased on the 14th day after modeling(P<0.05),but decreased on the 21st day(P<0.05);the protein and mRNA expressions of alpha-actin,transforming growth factor beta 1,and type Ⅰ collagen in the Lijin group were significantly decreased compared with those in the model group(P<0.05).Immunohistochemical results showed that the expression of type Ⅰ collagen in the Lijin group was significantly lower than that in the model group(P<0.05).To conclude,Lijin manipulation could improve the repair quality of skeletal muscle injury by inhibiting inflammation,promoting the proliferation and differentiation of muscle satellite cells,and reducing fibrosis.

Objective To screen and identify specific monoclonal antibodies targeting the surface envelope protein E8L of the monkeypox intracellular mature virus based on mRNA immunization and single-cell sequencing.Methods E8L-mRNA was synthesized in vitro and encapsulated into lipid nanoparticles(LNPs)for immunization to induce immune responses.The antibody level in blood serum was detected to find out when the mice had produced sufficient antibodies.B cells in the spleen of the mice were isolated by flow cytometry sorting,followed by library construction and sequencing.Based on the analysis of light and heavy chain sequences of the antibodies from the sequencing data,E8L candidate monoclonal antibodies were screened according to the abundance ranking and expressed.The affinity of these monoclonal antibodies was detected by enzyme-linked immunosorbent assay(ELISA).Candidate antibodies with high affinity for E8L were applied to immunochromatographic assays to detect the monkeypox virus(MPXV)E8L protein.Results Mice were immunized by lipid nanoparticles encapsulating E8L-mRNA.Approximately 1.5 million B cells were selected by flow cytometry.After single-cell sequencing,17 candidate monoclonal antibodies for E8L were identified and expressed.Two high affinity monoclonal antibodies were obtained for monkeypox virus E8L protein by ELISA.These two antibodies were used as the basis for immunochromatographic assays to detect E8L protein,with a sensitivity of 0.5 ng/mL.Conclusion Two monoclonal antibodies with high affinity for the MPXV E8L protein are obtained,which can be potentially used for detecting monkeypox virus.

BACKGROUND:Electroacupuncture can treat oligoasthenospermia by stimulating hormone synthesis and release in the hypothalamus,pituitary gland and testis,but its mechanism is complex,and the efficacy of electroacupuncture is closely related to the idea of acupoint compatibility.OBJECTIVE:To explore whether electroacupuncture therapy can improve the function status of hypothalamic-pituitary-testicular axis by adjusting the level of serum related sex hormones in rats,so as to treat adenine-induced oligoasthenospermia in rats.METHODS:Fifty adult male Sprague-Dawley rats were randomly divided into blank,model,electroacupuncture,L-carnitine,and non-meridian and non-acupuncture point groups,with 10 rats in each group.Except for the blank group,the rats in the other four groups were given 200 mg/(kg·d)adenine via intragastric administration for 28 days to establish oligoasthenospermia models.After modeling,the rats in the electroacupuncture group were given electroacupuncture treatment at Zhongji,Guanyuan,Zusanli and Sanyinjiao acupoints,the non-acupoint group was selected for electroacupuncture treatment at three non-acupoint points,and the rats in the L-carnitine group were given oral infusion of L-carnitine,once daily,for 28 continuous days.After treatment,the kidney and testicular organ coefficients were calculated.Sperm count,survival rate and motility rate were determined.Pathological morphological changes of kidney and testicular tissues were observed.Serum sex hormones testosterone,gonadotropin releasing hormone,follicle stimulating hormone,luteinizing hormone,estradiol,statin B and prolactin levels were detected in rats of each group.RESULTS AND CONCLUSION:(1)The renal organ coefficient in the model group was higher than that in the blank,electroacupuncture and L-carnitine groups(P＜0.05),the renal organ coefficient in the non-acupoint group was higher than that in the electroacupuncture group(P＜0.05),and there was no significant difference in the testicular organ coefficient among all groups(P＞0.05).(2)Compared with the blank group,sperm count,survival rate and motility rate were decreased in the model group(P＜0.05);compared with the model group,sperm count,survival rate and motility rate were increased in the electroacupuncture group and L-carnitine group(P＜0.05);compared with the non-acupoint group,sperm count,survival rate and motility rate were higher in the electroacupuncture group(P＜0.05).(3)Hematoxylin-eosin staining results indicated that the kidney and testicular tissues of rats in the model group were severely damaged,while damage to kidney and testicular tissues was less in the electroacupuncture group and L-carnitine group compared with the model group,but severer in the non-acupoint group than the electroacupuncture group.(4)Compared with the blank group,the levels of testosterone,gonadotropin releasing hormone,and inhibin B in the serum of rats in the model group were decreased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were increased(P＜0.05).Compared with the model group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin B in the serum of rats in the electroacupuncture group and L-carnitine group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).Compared with the non-acupoint group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin Bin the serum of the rats in electroacupuncture group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).To conclude,electroacupuncture at Zhongji,Guanyuan,Zusanli,and Sanyinjiao acupoints can effectively treat oligoasthenospermia by regulating the functional state of the hypothalamic-pituitary-testicular axis in rats.

Autoimmune oophoritis is a rare cause of primary ovarian insufficiency(POI), and some patients may also have concurrent adrenal insufficiency. This condition significantly impairs reproductive function and is often difficult to distinguish from common causes of POI, leading to missed or incorrect diagnoses. This paper reports a case of autoimmune oophoritis treated in our department and provides an updated summary of recent advances in the understanding of this condition, with the aim of raising clinical awareness and improving diagnostic accuracy among healthcare professionals.

BACKGROUND:Electroacupuncture can treat oligoasthenospermia by stimulating hormone synthesis and release in the hypothalamus,pituitary gland and testis,but its mechanism is complex,and the efficacy of electroacupuncture is closely related to the idea of acupoint compatibility.OBJECTIVE:To explore whether electroacupuncture therapy can improve the function status of hypothalamic-pituitary-testicular axis by adjusting the level of serum related sex hormones in rats,so as to treat adenine-induced oligoasthenospermia in rats.METHODS:Fifty adult male Sprague-Dawley rats were randomly divided into blank,model,electroacupuncture,L-carnitine,and non-meridian and non-acupuncture point groups,with 10 rats in each group.Except for the blank group,the rats in the other four groups were given 200 mg/(kg·d)adenine via intragastric administration for 28 days to establish oligoasthenospermia models.After modeling,the rats in the electroacupuncture group were given electroacupuncture treatment at Zhongji,Guanyuan,Zusanli and Sanyinjiao acupoints,the non-acupoint group was selected for electroacupuncture treatment at three non-acupoint points,and the rats in the L-carnitine group were given oral infusion of L-carnitine,once daily,for 28 continuous days.After treatment,the kidney and testicular organ coefficients were calculated.Sperm count,survival rate and motility rate were determined.Pathological morphological changes of kidney and testicular tissues were observed.Serum sex hormones testosterone,gonadotropin releasing hormone,follicle stimulating hormone,luteinizing hormone,estradiol,statin B and prolactin levels were detected in rats of each group.RESULTS AND CONCLUSION:(1)The renal organ coefficient in the model group was higher than that in the blank,electroacupuncture and L-carnitine groups(P＜0.05),the renal organ coefficient in the non-acupoint group was higher than that in the electroacupuncture group(P＜0.05),and there was no significant difference in the testicular organ coefficient among all groups(P＞0.05).(2)Compared with the blank group,sperm count,survival rate and motility rate were decreased in the model group(P＜0.05);compared with the model group,sperm count,survival rate and motility rate were increased in the electroacupuncture group and L-carnitine group(P＜0.05);compared with the non-acupoint group,sperm count,survival rate and motility rate were higher in the electroacupuncture group(P＜0.05).(3)Hematoxylin-eosin staining results indicated that the kidney and testicular tissues of rats in the model group were severely damaged,while damage to kidney and testicular tissues was less in the electroacupuncture group and L-carnitine group compared with the model group,but severer in the non-acupoint group than the electroacupuncture group.(4)Compared with the blank group,the levels of testosterone,gonadotropin releasing hormone,and inhibin B in the serum of rats in the model group were decreased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were increased(P＜0.05).Compared with the model group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin B in the serum of rats in the electroacupuncture group and L-carnitine group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).Compared with the non-acupoint group,the levels of testosterone,gonadotropin-releasing hormone,and inhibin Bin the serum of the rats in electroacupuncture group were increased(P＜0.05),and the levels of follicle-stimulating hormone,luteinizing hormone,estradiol,and prolactin were decreased(P＜0.05).To conclude,electroacupuncture at Zhongji,Guanyuan,Zusanli,and Sanyinjiao acupoints can effectively treat oligoasthenospermia by regulating the functional state of the hypothalamic-pituitary-testicular axis in rats.

Autoimmune oophoritis is a rare cause of primary ovarian insufficiency(POI), and some patients may also have concurrent adrenal insufficiency. This condition significantly impairs reproductive function and is often difficult to distinguish from common causes of POI, leading to missed or incorrect diagnoses. This paper reports a case of autoimmune oophoritis treated in our department and provides an updated summary of recent advances in the understanding of this condition, with the aim of raising clinical awareness and improving diagnostic accuracy among healthcare professionals.

BACKGROUND:Lijin manipulation can reduce fibrosis scar hyperplasia and promote skeletal muscle repair.However,improper activation of the Wnt/β-catenin signaling pathway can aggravate the fibrosis of injured skeletal muscle and adversely affect the repair process of skeletal muscle.To study the regulatory effect of Lijin manipulation on the Wnt/β-catenin signaling pathway is conducive to elucidate the related mechanisms of Lijin manipulation in reducing fibrosis scar hyperplasia and promoting skeletal muscle injury repair.OBJECTIVE:To explore the mechanism of Lijin manipulation in promoting the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group with 15 rabbits in each group.Gastrocnemius muscle percussion modeling was performed in both model group and Lijin group.Lijin manipulation was performed in the Lijin group on the 3rd day after modeling,once a day,15 minutes once.Five animals in each group were selected and killed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining and the content of collagen fiber was observed by Masson staining.Western blot was used to detect the protein expression of Wnt3a,β-catenin,GSK3β,p-GSK3β,TCF,type I collagen and type III collagen in gastrocnemius muscle,and RT-PCR was used to detect the mRNA expression of Wnt3a,β-catenin and TCF.The expression of β-catenin was detected by immunofluorescence,and the expression of type I collagen and type III collagen was detected by immunohistochemistry.RESULTS AND CONCLUSION:The results of hematoxylin-eosin staining and Masson staining showed that compared with the model group,inflammatory cell infiltration and collagen fiber amount decreased in the Lijin group(P<0.001),and muscle fibers gradually healed.Western blot results showed that compared with the model group,the protein expression levels of Wnt3a,β-catenin,TCF,type I collagen and type III collagen were significantly decreased in the Lijin group at all observation time points(P<0.05),while the ratio of P-GSK3β/GSK3β was significantly increased in the Lijin group at all observation time points compared with the model group(P<0.05).RT-PCR results showed that compared with the model group,the mRNA expression levels of Wnt3a,β-catenin and TCF were significantly decreased in the Lijin group at all observation time points(P<0.001).Immunofluorescence results showed that compared with the model group,the fluorescence intensity of β-catenin expression in the Lijin group was significantly decreased at each observation time point and gradually became similar to that in the blank group(P<0.001).Immunohistochemical results showed that the expression levels of type I collagen and type III collagen in the Lijin group were significantly lower than those in the model group(P<0.01).To conclude,Lijin manipulation could inhibit the abnormal activation of the Wnt/β-catenin signaling pathway,reduce fibrotic scar hyperplasia,and promote the repair of injured skeletal muscle.

BACKGROUND:Excessive apoptosis in skeletal muscle cells will destroy the dynamic balance of the number of myocytes,leading to pathological injury of skeletal muscle.Lijin manipulation is effective in treating skeletal muscle injury,but whether it can inhibit apoptosis and promote the repair of skeletal muscle injury is unknown.OBJECTIVE:To explore the mechanism by which Lijin manipulation reduces inflammation and apoptosis during the repair of skeletal muscle injury in rabbits.METHODS:Forty-five healthy adult Japanese white rabbits were randomly divided into blank group,model group and Lijin group(n=15 per group).No intervention was performed in the blank group.Gastrocnemius muscle percussion molding was performed in both the model group and Lijin group.After modeling,the model group was not treated,while the Lijin manipulation(Stroking,kneading,and rubbing)was performed in the Lijin group on the 3rd day,once a day,15 min/time.Sampling in each group was performed on the 7th,14th and 21st days after modeling.The general morphological structure of gastrocnemius was observed by hematoxylin-eosin staining.The ultrastructure of gastrocnemius muscle was observed by transmission electron microscopy.Apoptosis of gastrocnemius cells was observed by TUNEL staining.The expressions of interleukin-1β,interleukin-6 and interleukin-10 in gastrocnemius muscle were detected by ELISA.The protein expressions of BAX,BCL-2 and Caspase3 in gastrocnemius muscle were detected by western blot.The mRNA expression of BAX and BCL-2 was detected by RT-PCR.RESULTS AND CONCLUSION:(1)Hematoxylin-eosin staining results showed that compared with the model group,inflammatory cells decreased in number,myocyte amount increased,and muscle tissue gradually healed in the Lijin group at each observation point.(2)The results of transmission electron microscopy showed that compared with the model group,the arrangement of muscle fibers at each observation point in the Lijin group was gradually orderly,mitochondria were gradually complete,Z-line arrangement was gradually regular,and free ribosomes were gathered.(3)TUNEL staining results showed that compared with the model group,apoptosis rate in the Lijin group was gradually decreased at all observation points(P<0.05).(4)ELISA results showed that compared with the model group,the expression of interleukin-1β and interleukin-6 in the Lijin group continued to decrease(P<0.05),while the expression of interleukin-10 increased on the 7th day after modeling,and then showed a downward trend(P<0.05).(5)Western blot results showed that compared with the model group,the expression of BCL-2 protein/BAX protein in the Lijin group was significantly increased at each observational point(P<0.05).The protein expression of Caspase3 decreased significantly(P<0.001),and was gradually similar to that of the blank group.(6)RT-PCR results showed that compared with the model group,the mRNA expression level of BCL-2/BAX in the Lijin group was significantly higher at each observational point(P<0.05).To conclude,Lijin manipulation can inhibit inflammation,reduce apoptosis,and promote the repair of injured skeletal muscle.