Objective:To analyze the change of bone mineral density (BMD) in postmenopausal women,and to study the relationship between BMD and the estrogen,cytokines,and serum levels of bone metabolism.Methods: 135 postmenopausal women were divided into the OP group (n=54),Osteopenia group (n=43) and normal group (n=38) according to the results of lumbar spine bone mineral density (BMD) scan.To detected the concentration of serum estradiol (E2),bone alkaline phosphatase (BALP) and osteocalcin (BGP),tartrate resistant acid phosphatase-5b (TRAP-5b),interleukin-6 (IL-6),tumor necrosis factor alpha (TNF-α),transforming growth factor beta (TGF-β1) and IL-10 of research objects in the three groups.And to analyze the relationship between BMD and the levels of E2,BALP,BGP,TRAP-5b,IL-6,TNF-α,TGF-β1 and IL-10.Results: In the OP group and the Osteopenia group,BMD and the levels of E2,TGF-β1 and IL-10 were obviously lower than that in the normal group (P<0.05),and the levels of BALP,BGP,TRAP-5b,IL-6 and TNF-α were obviously higher than that in the normal group (P<0.05).When compared with the Osteopenia group,the BMD and E2 level in the OP group was more lower (P<0.05),and the levels of BALP,BGP,TRAP-5b,IL-6 and TNF-α were more higher (P<0.05).E2 and TGF-β1 showed the significant positive correction with BMD (P<0.05),and the BALP,BGP,TRAP-5b,IL-6 and TNF-α were all negatively corrected with BMD (P<0.05).However,no significant difference of relevance was seen between IL-10 and BMD (P>0.05).The low serum E2 levels and high serum levels of BALP,BGP,TRAP-5b,IL-6 and TNF-α were the independent factors affecting the reduction of BMD (P<0.05).Conclusion: The E2 level in patients with PMOP decreased obviously,which caused the imbalance of bone remodeling and inflammatory reaction,and which was related to the decrease of BMD.

Objective:To explore the effect on sex hormone functional of extracts of Herba Epimedii /Fructus Ligustri Lucidi in osteoporosis rats.Methods:50 cases of 3 months Wistar rats were rando mly divided into normal group ,model group,extract low dose group(extract-L),extract middle dose group(extract-M),extract high dose group(extract-H),10 cases in each group,in addition to the normal group,other 4 groups were bulided OP model by filling and feeding retinoic acid for 15 d,then normal group and model group were filling physiological saline;extract-L,extract-M,extract-H were respectively feeding extracts of Herba Epimedii /Fructus Ligustri Lucidi at 50 mg/kg,100 mg/kg,200 mg/kg,both for 3 weeks.Measured the bone mineral density (BMD),observed the femoral pathology change of left femur with hematoxylin and eosin ( HE ) staining, measured trabecular bone tissue morphology related parameters ,analyzed the hormone receptor protein expression with immunohistochemistry .Results:The BMD of the whole femur ,femur head,and femur neck in normal group were all significantly higher than that of model group ,the BMD of the whole femur ,femur head, and femur neck in extract-M and extract-H group were all significantly higher than that of model group ,the BMD of the whole femur in extract-L group was significantly higher than that of model group ,the differences were statistically significant (P<0.05 or P<0.01).The bone trabecular number had significantly increased ,integrity,continuity improved significantly ,bone histomorphometry such as Tb.Ar, Tb.Th,and Tb.N in model group were decreased ,and Tb.Sp was increased,extract-M and extract-H group had remarkable effect on each index ,while extract-L group only had a remarkable effect on Tb .Ar and Tb.Th ( P<0.05 or P<0.01 ) , compared with model group.The expression of AR , ER in normal groups were significantly higher than that of model group , the expression of AR , ER in extract-H group was significantly higher than that of model group , the differences were statistically significant ( P<0.05 or P<0.01 ) . Conclusion:This study demonstrated that the combination with TFE and TIFL had potential protective effects on osteoporosis rats induced by retinoic acid .The combination with TFE and TIFL is able to increase BMD ,reverse pathological changes of bone tissue ,co-ordinate hormones of the hypothalamic-pituitary-gonadal axis ,and upregulate the expression of sex hormone receptors .

Objective This study was to investigate the effect of caloric restriction on the plasticity of visual cortex in adult monocular deprivation (MD) amblyopic mice,as well as to promote the treatment of amblyopia,and to explore the possible molecular mechanism of this benefical effect.Methods Fifty healthy newborn Kunming mice of clean grade were randomly divided into 3 groups using a random number table method:normal control group (n =14),MD+ ad libitum group (n=18) and MD+ caloric restriction group (n=18).A mouse model of adult MD amblyopia was established,and caloric restriction intervention and ad libitum were performed on MD + caloric restriction group and MD+ ad libitum group,respectively.The visual acuity and flash visual evoked potential (F-VEP) of each group were detected.The synaptic structure of visual cortex neurons was observed by transmission electron microscope,and the expression of phosphorylated AMP-activated protein kinase-α(p-AMPKα) and silent information regulator 1 (SIRT1) in visual cortex were detected by Western blot.The animal feeding and use was in accordance with the standards set by the ARVO.Results The weight of mice in MD+ caloric restriction group increased from the beginning of the first week,and was significantly lower than that in the MD + ad libitum group (P＜0.05).Compared with the MD+ ad libitum group,the visual acuity was restored,the latency was shortened,and the amplitude of F-VEP was increased in the deprived eyes of MD+ caloric restriction group (all at P＜0.05).Transmission electron microscope observation showed that the width of synaptic gap of visual cortical neurons was significantly narrower,and the thickness of postsynaptic density was significantly thicker in MD+ caloric restriction group than that in the MD+ ad libitum group (both at P＜0.05);compared with the normal control group,the synaptic gap was widened and the postsynaptic density was significantly thicker than that in the MD+ ad libitum group (both at P＜0.05).Western blot showed that the expression of p-AMPKα in visual cortex in the normal control group,MD+ caloric restriction group and MD+ ad libitum group was 0.89±0.03,0.94±0.02 and 0.74 ±0.02,and the expression of SIRT1 was 0.97±0.11,0.95±0.14 and 0.58±0.13,respectively,showing significant differences among the three groups (F =14.57,P =0.00;F=23.91,P=0.00),the expressions of p-AMPKα and SIRT1 in visual cortex were increased in MD+ caloric restriction group than those in M D+ ad libitum group (both at P＜0.05).Conclusions Caloric restriction can restore the ultrastructure of synapses and improve the visual cortical plasticity in adult MD mice,so that help to improve visual function.Its mechanism may be related to the activation of AMPK-SIRT1 pathway.

Objective To examine role and possible mechanism of enriched environment (EE) on regulating recovery of visual function in adult monocular deprivation amblyopia mice.Methods A total of 72 healthyKunming mice were divided into normal control group,monocular deprivation (MD) group,MD+EE group and M D+ fluoxetine group by random number table.Except for the normal control group,the mice in the other groups were sutured on the right eyelid 21 days after birth to establish MD amblyopia model.the mice were fed in standard environment or EE for 4 weeks according to the group.Visual acuity and flash visual evoked potential (F-VEP) of mice in each group were detected.The distribution of microtubule associated protein 2 (MAP2) in visual cortex of adult amblyopic mice were detected by immunohistochemistry.The expression of MAP2,synaptophysin (SYP) and postsynaptic density protein-95 (PSD-95) protein in visual cortex of adult amblyopic mice were detected by western blot.The experimental protocol was approved by the Animal Care and Use Committee of Hunan Children's Hospital and conformed to the National Institutes of Health Guide for the Care and Use of Laboratory Animals.Results There was a significant difference in the visual acuity of deprived eye among each group (F=114.632,P＜0.001).The visual acuity in MD group is lower than that in normal control group,with a significant difference (t =15.480,P＜0.001).Compared with MD group,visual acuity was restored in MD+ EE group and MD +fluoxetine group,with significant differences (t =15.071,P ＜ 0.001;t =14.841,P ＜ 0.001).There was a significant difference in the P2 latency and amplitude of F-VEP in deprived eye among each group (F=36.510,P=0.000;F=34.140,P=0.000).Compare with normal control group,P2 latency was prolonged and P2 amplitude of F-VEP was decreased in deprived eye in MD group,with significant differences (t =10.220,P =0.000;t =10.09,P =0.000).Western blot assay showed that there was a significant difference in the expression of MAP2 in visual cortex contralateral deprived eye among each group (F=18.142,P=0.000).The expression of MAP2 in MD group was significantly lower than that in normal contral group (t=3.056,P＜0.01);Compared with MD group,MAP2 expression was increased in MD+EE group and MD+fluoxetine group (t =2.541,P =0.031;t =2.157,P =0.017).There were significant differences in the expression of SYP and PSD-95 in visual cortex contralateral to deprived eye among each group (F =12.871,P =0.000;F =25.060,P =0.000).Compared with normal contral group,SYP and PSD-95 expression in visual cortex were down-regulated in MD group,with significant differences (t =6.054,P =0.000;t =8.631,P =0.000).The expression of SYP and PSD-95 protein in MD+EE group and MD+fluoxetine group were significantly higher than those in MD group (all at P＜0.05).Conclusions EE can recover visual function through up-regulating the expression of MAP2,which can modulate the dendritic branch trim and neural plasticity of visual cortex in adult MD mice.

Rheumatoid arthritis belongs to arthralgia syndrome in the theory of traditional Chinese medicine， and cold-dampness obstruction syndrome and dampness-heat obstruction syndrome are core syndromes and main syndrome differentiation types of this disease. Fine therapeutic effects have been obtained in the long-term clinical practice of many famous traditional Chinese medicine practitioners following the syndrome differentiation and treatment based on the guiding principles of cold and heat. To adapt to the clinical diagnosis practice of combining disease differentiation and syndrome differentiation， and to better carry out basic research on integrated Chinese and Western medicine and preclinical study on new traditional Chinese medicines， Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-Dampness Obstruction Syndrome and Dampness-Heat Obstruction Syndrome （hereinafter referred to as the Guidelines） were compiled by our research group， in cooperation with the renowned experts in research fields including traditional Chinese medicine， clinical medicine， zoology and evidence-based medicine， which provide a meaningful reference for scientific research， teaching and clinical applications. The compilation process of the Guidelines was guided by the theory of disease and syndrome integration and the principles of "evidence takes the main place， consensus plays an auxiliary role， and experience serves as the reference". Based on the comprehensive evaluation of pathogenesis homology， behavioral phenotypic consistency， and drug treatment predictability compared between animal models and human diseases， by the nominal group method， "recommendations" were formed for recommendations supported by evidence， and "consensus recommendations" were formed for recommendations not supported by evidence. Guidelines were formed involving content such as animal types， arthritis modeling methods， external stimulation conditions， and modeling assessment indicators during the establishment of the animal models of rheumatoid arthritis with cold-dampness obstruction syndrome and dampness-heat obstruction syndrome. The Guidelines are applicable for the disease and syndrome research on rheumatoid arthritis， investigation of therapeutic mechanisms， and development of new traditional Chinese medicine. The Guidelines also provide a reference for the establishment of guidelines on other types of diseases and syndromes combined with animal models to further promote the modernization of traditional Chinese medicine research and its integration with international academic development.

The Guidelines for Establishing Animal Models of Rheumatoid Arthritis with Cold-dampness Obstruction Syndrome and Dampness-heat Obstruction Syndrome （hereinafter referred to as the Guidelines） （No. T/CACM1567-2024） was published by Chinese Association of Chinese Medicine on January 11， 2024. To assist researchers and medical workers in understanding and applying the Guidelines more accurately， and also to provide reference and assistance for the establishment of guidelines on other types of diseases and syndromes combined with animal models， this paper made a declaration of the workflow， technological links， development references， promotion of its application and after-effect evaluation of the Guidelines that has been made according to the requirements of "Draft Group Standard of the Standardization Office of the Chinese Association of Traditional Chinese Medicine".