1.Regulatory mechanism of electroacupuncture on hypothalamic-pituitary-testicular axis in oligospermic rats
Zhenning ZHAO ; Kaiying LI ; Nan YANG ; Wenjing SUN ; Xiaoge WEI ; Jing MU ; Huisheng MA
Chinese Journal of Tissue Engineering Research 2025;29(26):5563-5571
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.
2.Mechanism of Lijin manipulation regulating scar formation in skeletal muscle injury repair in rabbits
Kaiying LI ; Xiaoge WEI ; Fei SONG ; Nan YANG ; Zhenning ZHAO ; Yan WANG ; Jing MU ; Huisheng MA
Chinese Journal of Tissue Engineering Research 2025;29(8):1600-1608
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.
3.Lijin manipulation alleviates fibrosis of injured skeletal muscle in rabbits
Kaiying LI ; Xiaoge WEI ; Zhenning ZHAO ; Fei SONG ; Nan YANG ; Yan WANG ; Jing MU ; Huisheng MA
Chinese Journal of Tissue Engineering Research 2025;29(14):2914-2921
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.
4.Regulatory mechanism of Lijin manipulation on inflammation and apoptosis in rabbits undergoing skeletal muscle injury repair
Kaiying LI ; Xiaoge WEI ; Fei SONG ; Zhenning ZHAO ; Nan YANG ; Hongyu LI ; Lili ZHAO ; Yan WANG ; Jing MU ; Huisheng MA
Chinese Journal of Tissue Engineering Research 2025;29(20):4215-4222
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.
5.Lijin manipulation alleviates fibrosis of injured skeletal muscle in rabbits
Kaiying LI ; Xiaoge WEI ; Zhenning ZHAO ; Fei SONG ; Nan YANG ; Yan WANG ; Jing MU ; Huisheng MA
Chinese Journal of Tissue Engineering Research 2025;29(14):2914-2921
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.
6.Regulatory mechanism of Lijin manipulation on inflammation and apoptosis in rabbits undergoing skeletal muscle injury repair
Kaiying LI ; Xiaoge WEI ; Fei SONG ; Zhenning ZHAO ; Nan YANG ; Hongyu LI ; Lili ZHAO ; Yan WANG ; Jing MU ; Huisheng MA
Chinese Journal of Tissue Engineering Research 2025;29(20):4215-4222
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.
7.Regulatory mechanism of electroacupuncture on hypothalamic-pituitary-testicular axis in oligospermic rats
Zhenning ZHAO ; Kaiying LI ; Nan YANG ; Wenjing SUN ; Xiaoge WEI ; Jing MU ; Huisheng MA
Chinese Journal of Tissue Engineering Research 2025;29(26):5563-5571
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.
8.Diagnostic value of cervical cell DNA ploidy analysis combined with B7-H4 and PKCδ for cervical cancer
Ningning ZHANG ; Zhe YANG ; Limei TAN ; Zhenning LI ; Di WANG ; Yongzhi WEI
Journal of International Oncology 2024;51(5):286-291
Objective:To investigate the diagnostic value of cervical cell DNA ploidy analysis combined with negative costimulatory molecule B7 homolog 4 (B7-H4) and protein kinase Cδ (PKCδ) for cervical cancer.Methods:A total of 160 cervical cancer patients diagnosed and treated at Shijiazhuang People's Hospital from January 2018 to January 2022 were selected as the cervical cancer group. Meantime, 160 women who were screened for cervical cancer in our hospital during this period were selected as the control group. According to the examination results, they were divided into normal or inflammatory group ( n=52), low-grade cervical intraepithelial neoplasia (CIN) group ( n=68) and high-grade CIN group ( n=40). The automatic cell image analysis system was used to analyze the DNA ploidy of cervical cells. The levels of B7-H4 and PKCδ in serum were determined by enzyme-linked immunosorbent assay. Pearson method was used to analyze the correlation between serum B7-H4 and PKCδ; the diagnostic value of cervical cell DNA ploidy analysis combined with serum B7-H4 and PKCδ in cervical cancer was evaluated by the receiver operator characteristic (ROC) curve; multivariate logistic regression was used to analyze the risk factors of cervical cancer. Results:The numbers of DNA ploidy positive cases of cervical cells in normal or inflammatory group, low-grade CIN group, high-grade CIN group and cervical cancer group were 16 (30.8%), 27 (39.7%), 26 (65.0%) and 127 (79.4%), respectively, with a statistically significant difference ( H=55.86, P<0.001). Further pin-by-pair comparison showed that compared with normal or inflammatory groups, the proportion of DNA ploidy positive in high-grade CIN group and cervical cancer group were higher (both P<0.05). The proportion of DNA ploidy positive in cervical cancer group was higher than that in low-grade CIN group ( P<0.05). Serum B7-H4 levels in normal or inflammatory group, low-grade CIN group, high-grade CIN group and cervical cancer group were (57.21±10.21), (79.17±11.34), (92.73±15.36), (126.56±20.25) ng/ml, respectively, with a statistically significant difference ( F=285.45, P<0.001). Serum PKCδ levels were (89.34±18.29), (71.79±15.82), (53.39±11.84), (40.23±10.21) ng/ml, respectively, with a statistically significant difference ( F=216.28, P<0.001). Further pin-by-pair comparison showed that serum B7-H4 levels in normal or inflammatory groups, low-grade CIN group, high-grade CIN group and cervical cancer group increased in turn (all P<0.05). Serum PKCδ levels in normal or inflammatory groups, high-grade CIN group and cervical cancer group were decreased in turn (all P<0.05). Pearson correlation analysis showed that the serum B7-H4 and PKCδ levels in patients with cervical cancer were negatively correlated ( r=-0.47, P<0.001). ROC curve analysis showed that the area under the curve (AUC) of cervical cell DNA ploidy for cervical cancer diagnosis was 0.82 (95% CI: 0.78-0.86), and the sensitivity and specificity were 83.9% and 79.9%, respectively. The AUC of serum B7-H4 in the diagnosis of cervical cancer was 0.92 (95% CI: 0.89-0.95), the sensitivity and specificity were 95.7% and 76.1%, respectively, and the cutoff value was 111.12 ng/ml. The AUC of serum PKCδ for diagnosis of cervical cancer was 0.92 (95% CI: 0.89-0.95), the sensitivity and specificity were 85.6% and 88.9%, respectively, and the cut-off value was 54.83 ng/ml. The AUC of the combined diagnosis of cervical cancer was 0.99 (95% CI: 0.97-0.99), and the sensitivity and specificity were 98.3% and 75.9%, respectively. The AUC of the combined diagnosis of cervical cancer was higher than that of DNA ploidy ( Z=8.00, P<0.001), serum B7-H4 ( Z=4.34, P<0.001), and serum PKCδ ( Z=4.61, P<0.001) alone. Multivariate logistic regression analysis showed that high level of B7-H4 in serum ( OR=2.94, 95% CI: 1.78-4.84, P<0.001), low level of PKCδ ( OR=4.33, 95% CI: 1.88-10.00, P=0.001) and positive DNA ploidy in cervical cells ( OR=5.77, 95% CI: 2.38-13.99, P<0.001) were independent risk factors for cervical cancer. Conclusion:The positive proportion of DNA ploidy in cervical cells of patients with cervical cancer is increased, the serum B7-H4 level is increased, the PKCδ level is decreased, and cervical cell DNA ploidy analysis combined with serum B7-H4 and PKCδ has a high diagnostic value for cervical cancer.
9.Predictive value of early indicators changes in blood test on the prognosis of patients with acute paraquat poisoning
Linlin CHAO ; Zhenning LIU ; Min ZHAO ; Mengying YANG ; Yongzhuang YE ; Guigui WEI
Chinese Critical Care Medicine 2020;32(6):732-736
Objective:To investigate the predictive value of early indicators changes in blood test on the prognosis of patients with acute paraquat poisoning.Methods:The clinical data of patients with acute paraquat poisoning admitted to emergency department of Shengjing Hospital of China Medical University from January 2012 to June 2019 were retrospectively analyzed. The changes of blood test indexes within 24 hours after admission were collected, including white blood cell count (ΔWBC), neutrophils count (ΔNE), lymphocytes count (ΔLY), monocytes count (ΔMO), arterial partial pressure of oxygen (ΔPaO 2), arterial partial pressure of carbon dioxide (ΔPaCO 2), arterial blood pH (ΔpH), bicarbonate radical (ΔHCO 3-), base excess (ΔBE), lactate (ΔLac), total protein (ΔTP), albumin (ΔALB), alanine aminotransferase (ΔALT), aspartate aminotransferase (ΔAST), total bilirubin (ΔTBil), direct bilirubin (ΔDBil), blood urea nitrogen (ΔBUN), serum creatinine (ΔSCr), serum calcium concentration (ΔCa 2+), and serum potassium concentration (ΔK +). Multivariate Logistic regression was used to analyze the risk factors of prognosis in patients with acute paraquat poisoning, and receiver operating characteristic (ROC) curve was drawn to analyze the predictive value of ROC curve for the death of patients with paraquat poisoning. Results:A total of 251 patients with acute paraquat poisoning were included, with 99 cases dead, and the mortality was 39.4%. The increase of the markers including ΔWBC, ΔLac, ΔALT, ΔAST, ΔTBil, ΔDBil, ΔBUN, ΔSCr and ΔK + within 24 hours of admission in the death group were significantly higher than that in the survival group; the decrease of the markers including ΔPaCO 2, ΔHCO 3-, ΔBE, ΔTP, and ΔALB in the death group were significantly greater than those in the survival group. The variables with statistical significance in the above single factor analysis were included in the multivariate Logistic regression analysis. The results showed that ΔLac, ΔSCr and ΔK + were independent risk factors for the prognosis of patients with acute paraquat poisoning [odds ratio ( OR) and 95% confidence interval (95% CI) were 1.662 (0.997-2.772), 1.045 (1.010-1.083) and 4.555 (1.190-17.429), respectively, all P < 0.05]. The area under the ROC curve (AUC) of ΔLac, ΔSCr and ΔK + for predicting death of patients with acute paraquat poisoning was 0.639 (95% CI was 0.505-0773), 0.811 (95% CI was 0.704-0.917), and 0.649 (95% CI was 0.519-0.779), respectively. When the cut-off of ΔLac was 1.85 mmol/L, the sensitivity was 87.9%, the specificity was 47.7%, and the diagnostic accuracy was 70.2%; when the cut-off of ΔSCr was 37.75 μmol/L, the sensitivity was 84.4%, the specificity was 77.9%, and the diagnostic accuracy was 80.5%; when the cut-off of ΔK + was 0.42 mmol/L, the sensitivity was 36.6%, the specificity was 90.7%, and the diagnostic accuracy was 68.3%. The efficiency of combination of ΔLac, ΔSCr, and ΔK + was greater than a single indicator in predicting death of patients with acute paraquat poisoning, with AUC of 0.911, and 95% CI of 0.834-0.989. Conclusions:ΔLac, ΔSCr, ΔK + within 24 hours of admission were all independent risk factors for the prognosis of patients with acute paraquat poisoning. ΔSCr > 37.75 μmol/L within 24 hours of admission would predict a poor prognosis in the patients with acute paraquat poisoning. Combined analysis of ΔLac, ΔSCr, and ΔK + can predict the prognosis of paraquat poisoning patients more accurately than single index.
10.Experimental study of the effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment
Zhenduo SHI ; Zhenning WEI ; Lin HAO ; Kun PANG ; Jiahe ZHOU ; Bingzheng DONG ; Zhiguo ZHANG ; Yan ZHAO ; Yufeng SUN ; Conghui HAN
Chinese Journal of Radiological Medicine and Protection 2020;40(8):573-581
Objective:To investigate the effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on targeted therapy of prostate cancer and its effect on tumor microenvironment. Methods:125I-RSOAds-hTERT/PSA ( 125I-virus complex) oncolytic adenovirus was constructed by PCR amplification and double restriction enzyme ligation. TUNEL staining, flow cytometry and Caspase-3 immunoblotting assay were used to detect the killing effect of 125I-RSOAds-hTERT/PSA oncolytic adenovirus on prostate cancer cells in vitro and in vivo, respectively. To explore the effect of 125I-virus complex on tumor tissue cytokine secretion levels, interleukin 2 (IL-2), IL-10, tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in the culture supernatant of human prostate cancer cell line PC3, mouse prostate adenocarcinoma cell line RM-1, and mice serum were detected by ELISA. We explored the regulation of 125I-virus complex on the expression of CD24, CD44 and prostate stem cell antigen (PSCA) in prostate tumor tissues and tumor cells through immunohistochemistry. Meanwhile, the expression levels of CD32 and vascular endothelial growth factor (VEGF), as well as CD4+ , CD8+ and macrophage infiltration in tumor tissue were detected through immunofluorescence experiments. Results:125I-virus complex oncolytic adenovirus significantly increased tumor cell apoptosis in vitro and in vivo that was significantly higher than that of 125I group and virus complex group. Meanwhile, IL-2 ( t=-183.30, -38.20, P<0.05), IL-10 ( t=113.80, 92.71, P<0.05), TNF-α ( t=-73.20, -73.91, P<0.05), IFN-γ ( t=-65.37, -139.70, P<0.05) increased in vitro and in vivo. 125I-virus complex reduced the expression of CD24, CD44 and PSCA in tumor cells and tumor tissue, reduced the weight of tumor tissue, inhibited angiogenesis of tumor tissue ( t=8.55, P<0.05), and regulated the immune response in tumor tissue. Conclusions:125I-virus complex targeting prostate cancer can significantly kill cancer cells, reduce the weight and angiogenesis of tumor, and improve tumor microenvironment.

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