Osteoporosis is a common bone disease， whose incidence is still on the rise， posing great challenges to patients and society. This review mainly studies the pathogenesis of osteoporosis from the aspects of oxidative stress， inflammatory response， and glucolipotoxicity-induced injury and clarifies the efficacy and mechanism of some active ingredients of traditional Chinese medicine against osteoporosis through the integration of in vitro and in vivo experiments. The experimental results suggest that some active ingredients can improve bone resorption markers and maintain bone homeostasis by modulating inflammation， oxidative stress， etc. These active ingredients regulate osteoporosis through the receptor activator of nuclear transcription factor-κB （NF-κB） ligand （RANKL） pathway， osteoprotegerin （OPG） pathway， Wnt/β-catenin pathway， NF-κB pathway， mitogen-activated protein kinase （MAPK） pathway， adenosine monophosphate （AMP）-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） pathway， and oxidative stress pathway. This review provides ideas for the progress of the prevention and treatment of osteoporosis with the active ingredients of traditional Chinese medicine， aiming to provide new potential lead compounds and reference for the development of innovative drugs and clinical therapy for the treatment of osteoporosis.

OBJECTIVE To promote the standardization of the “Indications” section in package inserts for Chinese patent medicines and ensure rational clinical and patient use. METHODS The “Function and Indications” information of package inserts for Chinese patent medicines was retrieved and collected from the 2020 edition of the Chinese Pharmacopoeia （Volume Ⅰ） and various national and regional standards. Identification criteria were established for syndrome， pathogenesis， disease name， and symptom terminology in the “Indications” section. Microsoft Office Access 2021 was utilized to create query tables for manual extraction of terminological elements， followed by the construction of a three-tier classification system for “Indications” descriptions. A standardized template for “Indications” was developed through quantitative analysis. RESULTS & CONCLUSIONS A total of 9 851 valid package inserts for Chinese patent medicines were included. Among these， the majority （7 991） contained symptom terminology， followed by disease names （5 867） and pathogenesis descriptions （5 167）. Within disease name terminology， Western medical disease names predominated （4 446）， followed by traditional Chinese medicine disease names （2 018）. The “Function and Indications” content of 6 962 package inserts complied with existing requirements. Notably， the secondary classifications of “disease name”， as well as the tertiary classification of “disease name+symptoms” and “symptoms”， failed to meet established standards. Two standardized templates for “Indications” were formulated based on pathogenesis and syndrome：“pathogenesis+disease name+symptoms” and “disease name+syndrome+symptoms”. The “Indications”section should provide complete and accurate information， adhere to standardized formatting， and employ appropriate conjunctions and punctuation. For non-prescription patent medicines， package inserts should be categorized into professional and patient versions. These measures will facilitate the standardization of “Indications” descriptions and advance the overall package inserts for Chinese patent medicines documentation.

Traditional Chinese Medicine (TCM), as a treasure of the Chinese nation, plays a significant role in maintaining public health. In 2019, the Central Committee of the Communist Party of China and the State Council proposed for the first time the establishment of a TCM registration and evaluation evidence system that integrates TCM theory, "personal experience" and clinical trials (referred to as the "Three-in-One" System) to promote the inheritance and innovation of TCM. Subsequently, the National Medical Products Administration issued several guiding principles to advance the improvement and implementation of this system. Owing to the complexity of its implementation, there are still differing understandings within the TCM industry regarding the positioning of the "Three-in-One" Registration and Evaluation Evidence System, as well as the connotation and value orientation of the "personal experience." To address this, Academician WANG Qi, President of the TCM Association, China International Exchange and Promotion Association for Medical and Healthcare and TCM master, led a group of academicians, TCM masters, TCM pharmacology experts and clinical TCM experts to convene a "Seminar on Promoting the Implementation of the ′Three-in-One′ Registration and Evaluation Evidence System for Chinese Medicinals." Through extensive discussions, an expert consensus was formed, clarifying the different roles of the TCM theory, "personal experience" and clinical trials within the system. It was further emphasized that the "personal experience" is the core of this system, and its data should be derived from clinical practice scenarios. In the future, the improvement of this system will require collaborative efforts across multiple fields to promote the high-quality development of the Chinese medicinal industry.

Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.
Animals ; Mice ; SARS-CoV-2/genetics* ; COVID-19 ; Vaccines, Synthetic/genetics* ; Genes, Reporter ; Bacteriophages ; Mice, Inbred BALB C

[摘 要] 目的：探讨贝母素乙对结肠癌HCT116细胞增殖的抑制作用及其分子机制。方法：利用不同浓度的贝母素乙处理人结肠癌细胞HCT116和正常结肠上皮细胞CCD841 CON，通过CCK-8法和结晶紫染色法检测贝母素乙对HCT116和CCD841 CON细胞增殖活力的影响，流式细胞术和WB法检测贝母素乙对HCT116细胞周期及其细胞周期相关蛋白表达的影响。构建HCT116移植瘤裸鼠模型和AOM/DSS结肠癌小鼠模型，观察贝母素乙对小鼠模型肿瘤生长和OS的影响，免疫组化法和WB法检测对移植瘤或肿瘤组织中细胞周期相关蛋白CDK4、CDK6和cyclin D1表达的影响。结果：贝母素乙可显著抑制结肠癌HCT116细胞的增殖能力（P<0.01），诱导HCT116细胞周期G0/G1期阻滞（P<0.01），降低CDK4、CDK6和cyclin D1的蛋白表达水平（均P<0.01）。荷瘤小鼠实验结果显示，贝母素乙（0.75 mg/kg）显著抑制HCT116细胞移植瘤的生长并延长荷瘤裸鼠的OS（P<0.05或P<0.01），降低AOM/DSS模型小鼠的体质量、延长OS、减少癌变肠组织的肿瘤个数和肿瘤体积，下调肿瘤组织中CDK4、CDK6和cyclin D1的蛋白表达（P<0.01或P<0.05）。结论：贝母素乙通过下调CDK4、CDK6和cyclin D1的表达水平，引起细胞周期G0/G1期阻滞，从而抑制结肠癌HCT116细胞的增殖。

BACKGROUND:Studies have shown that insulin-like growth factor 1/platelet-derived growth factor has an inhibitory effect on fibroblast apoptosis.miR-141-3p in bone marrow stromal cells increases with age and has a relationship with the activation of inflammatory signaling pathways,suggesting that it may be a therapeutic target for lumbar disc herniation. OBJECTIVE:To explore the effects of miR-141-3p on dorsal root ganglion inflammation and lower limb pain in rats with lumbar disc herniation by regulating insulin-like growth factor 1/platelet-derived growth factor. METHODS:Fifty male Sprague-Dawley rats,SPF level,were randomly divided into normal group,model group,miR-NC group,miR-141-3p inhibitor group and miR-141-3p mimics group,with 10 rats in each group.Except for the normal group,animal models of lumbar disc herniation were established in rats by autologous nucleus pulposus transplantation.After successful modeling,rats in the miR-NC,miR-141-3p inhibitor and miR-141-3p mimics groups were injected intrathecally with 10 μL of 20 μmol/L miR-NC,miR-141-3p inhibitor,miR-141-3p mimics,once a day for 28 days,respectively,while those in the normal and model groups were injected with the same volume of saline at the same location at the same time.Paw withdrawal thermal latency threshold was used to evaluate lower limb pain in rats.The mRNA expression of miR-141-3p in dorsal root ganglion tissue was detected by real-time fluorescence quantitative PCR,the levels of inflammatory factors in dorsal root ganglion tissue were detected by ELISA,and the expression of insulin-like growth factor 1/platelet-derived growth factor in dorsal root ganglion tissue was detected by western blot.The correlation between miR-141-3p and insulin-like growth factor 1/platelet-derived growth factor was analyzed. RESULTS AND CONCLUSION:There were no significant differences in all indexes between the miR-NC group and the model group.Paw withdrawal thermal latency threshold was significantly lower in the model group than in the normal group(P＜0.05),significantly lower in the miR-141-3p inhibitor group than the miR-NC group(P＜0.05),and significantly higher in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The mRNA expression of miR-141-3p in dorsal root ganglion tissue was significantly lower in the model group than in the normal group(P＜0.05),significantly lower in the miR-141-3p inhibitor group than in the miR-NC group(P＜0.05),and significantly higher in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The levels of tumor necrosis factor α,interleukin 1β,and interleukin 1 in dorsal root ganglion tissue were significantly higher in the model group than in the normal group(P＜0.05),significantly higher in the miR-141-3p inhibitor group than in the miR-NC group(P＜0.05),and significantly lower in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The protein expressions of insulin-like growth factor 1 and platelet-derived growth factor in dorsal root ganglion tissue were significantly lower in the model group than in the normal group(P＜0.05),significantly lower in the miR-141-3p inhibitor group than in the miR-NC group(P＜0.05),and significantly higher in the miR-141-3p mimics group than in the miR-141-3p inhibitor group(P＜0.05).The expressions of insulin-like growth factor 1 and platelet-derived growth factor showed a positive correlation with miR-141-3p(r=0.904,P＜0.001;r=0.879,P＜0.001).To conclude,miR-141-3p can significantly improve lower limb pain and inhibit inflammation in dorsal root ganglia in rats with lumbar disc herniation,and its mechanism may be related to the promotion of insulin-like growth factor 1/platelet-derived growth factor expression.

OBJECTIVE To predict potential quality markers(Q-markers) in Yinhua Miyanling tablets based on fingerprinting and network pharmacology methods. METHODS HPLC fingerprints of 13 batches of Yinhua Miyanling tablets were established, and the similarity analysis was carried out using the "Chromatographic Fingerprint Evaluation System for Traditional Chinese Medicine" to identify the common peaks and attribute them. The fingerprints of Yinhua Miyanling tablets were investigated using chemometrics, cluster analysis, principal component analysis and orthogonal partial least squares discriminant analysis in combination with SPSS 26.0 and SIMCA 14.1 software to identify the major signature components responsible for the differences. The network pharmacology was used to screen and analyze the targets and pathways of Yinhua Miyanling tablets, construct a "drug-component-target-pathway" network diagram, and predict the Q-Marker and core targets of Yinhua Miyanling tablets. RESULTS HPLC fingerprint of Yinhua Miyanling tablets was established, and 27 common peaks including chlorogenic acid, mangostin, wild baicalin, lignocerin and quercetin were identified. Chemical pattern recognition analysis screened five components as differential markers for Yinhua Miyanling tablets. Five active ingredients, 20 core targets and 20 key pathways were screened by network pharmacology, showing that all five active ingredients could be used as potential Q-Markers. CONCLUSION The method is stable, accurate and feasible for screening five chemical components as potential Q-Markers for Yinhua Miyanling tablets. It provides a reference for the overall control of the quality of Yinhua Miyanling tablets, and also lays the foundation for further research on the mechanism of action of Yinhua Miyanling tablets.

Objective To establish an allogeneic rat model of endometriosis and to evaluate the effects of gonadotropin-releasing hormone (GnRH) agonist GenSci006 on experimental rat endometriosis. Methods Endometrium from SPF grade donor female SD rats were transplanted onto the abdominal wall of recipient female rats to construct an allogeneic endometriosis model. The rats undergoing sham surgery were divided into the sham group. Three weeks later, the length, width and height of the ectopic endometrium were measured, and the volume of the endometrium (V₁) was calculated before drug administration. The modeling rats were randomly divided into four groups: model group, triptorelin group (0.25 mg/kg), GenSci006-1 group (0.125 mg/kg) and GenSci006-2 group (0.25 mg/kg). Each group had 16 rats and received a single dose of the corresponding drug. The sham group and model group were administered an equal volume of solvent. Three weeks after administration, ectopic endometrium was measured to calculate the volume V₂ and inhibition rate. The effect of GenSci006 on rat uterus and ovarian tissues was assessed by comparing organ coefficients and changes in pathological sections. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of serum estradiol (E₂), progesterone (P₄), follicle stimulating hormone (FSH), and luteinizing hormone (LH). Real-time fluorescent quantitative PCR was used to detect the expression of GnRH receptor (GnRHR) mRNA in the hypothalamus and pituitary. Western blot was used to detect the expression of estradiol receptor alpha (ERα), beta (ERβ) and progesterone receptor (PR) in ectopic endometrium. Results Three weeks after administration, compared with the model group, the body weight of rats in the triptorelin and GenSci006-2 groups significantly increased (P < 0.05), while the volume of ectopic endometrium significantly decreased (P < 0.05). Compared with the sham group, the model group showed no significant changes in uterine and ovarian organ coefficients or endometrial thickness (P > 0.05). Compared with the model group, the uterine organ coefficients and endometrial thickness were significantly reduced in the triptorelin and GenSci006-2 groups (P < 0.05). Compared with the sham group, the serum levels of E₂, P₄, FSH and LH in the model group showed no significant changes (P > 0.05). Compared with the model group, the ovarian organ coefficient and serum P₄ levels of rats in the Triptorelin, GenSci006-1, and GenSci006-2 groups were significantly reduced (P < 0.05), while the serum LH levels of rats in the GenSci006-1 group were significantly increased (P < 0.05). However, there were no significant changes in serum E₂ and FSH levels in each group (P > 0.05). Compared with the model group, the expression levels of GnRHR mRNA in the pituitary tissue of rats in the triptorelin and GenSci006-2 groups were significantly downregulated (P < 0.05), with no significantly changes in the hypothalamus (P > 0.05). There were no significant changes in the expression level of GnRHR mRNA in the hypothalamus or the protein levels of ERα, ERβ and PR in the ectopic endometrial tissue in any group (P > 0.05). Conclusion The allogeneic endometriosis rat model is a suitable animal model for screening and evaluating drugs for treating endometriosis. The volume of ectopic endometrium, inhibition rate, uterine and ovarian organ coefficients, and serum E₂ levels may serve as indicators for detecting drug efficacy.

Background In recent years, Hefei has experienced rapid industrial development, and the concentration of ozone (O₃) in the environment has increased year by year. Ambient O₃ pollution has become one of the big issues in Hefei. Objective To explore the health risks of non-accidental deaths (NAD) in local residents associated with ambient O₃ in Hefei City, and propose effective measures. Methods The data of daily NAD, mortality due to circulatory system diseases, mortality due to respiratory system diseases, atmospheric pollution, and meteorological factors were collected in Hefei from 2017 to 2020. By fitting daily average temperature, relative humidity, time trend, and seasonal trend, a generalized additive model was applied to evaluate the single-day effects (lag0-lag5) and cumulative lag effects (lag01-lag05) of NAD, circulatory system mortality, and respiratory system mortality among residents of Hefei City for every 10 μg·m⁻³ increase in the daily maximum 8-hour average ozone (O_{3-8 h}) concentration. The effects of O_{3-8 h} concentration on total NAD were also examined by stratification of age and sex. Results During the period of 2017 to 2020, the average concentration of O_{3-8 h} in Hefei City was 99.12 μg·m⁻³, and the total number of non-accidental deaths was 45787. For the O_{3-8 h} concentration per 10 μg·m⁻³ increase, the single-day effect and cumulative lag effect on NAD among residents reached their maximum values at lag0 and lag02, respectively, with the NAD increasing by 0.46% (95%CI: 0.06%, 0.87%) and 0.76% (95%CI: 0.27%, 1.26%); the maximum estimates for deaths from circulatory diseases appeared on lag03 increasing by 0.78% (95%CI: 0.08%, 1.48%). The stratified analyses results showed that the single-day effects on the NAD in males and females reached the maximum values on lag1 and lag0, increasing by 0.64% (95%CI: 0.21%, 1.08%) and 0.61% (95%CI: 0.03%, 1.20%), respectively; on lag02, the NAD among males was the highest, reaching 0.98% (95% CI: 0.39%, 1.57%); significant estimates for NAD among residents aged 65 years and above were observed on lag1 and lag02 rising by 0.41% (95%CI: 0.02%, 0.81%) and 0.67% (95%CI: 0.13%, 1.21%), respectively. After adjusting the degrees of freedom of time trend and other atmospheric pollutants, there was no statistically significant change in the risk of NAD death among residents associated with O_3-8h concentration. Conclusion The increase O₃ concentration in Hefei is correlated with an increased NAD risk among residents, with males and people over 65 years old having a higher risk of death due to O₃ exposure. It is necessary for the government to play an important role in developing prevention and control measures to reduce the excess risk of death associated with O₃ exposure.

OBJECTIVES: To investigate the cerebral metabolism in the patients with type 2 diabetes mellitus-associated cognitive dysfunction (T2DACD) and explore the mechanism of electroacupuncture (EA) at the acupoints for Tiaozang Xingshen (adjusting zangfu function and rescuing the spirit) in treatment of T2DACD, using magnetic resonance spectroscopy. METHODS: Fifteen patients with T2DACD (observation group) and 22 healthy subjects (control group) were enrolled. In the observation group, the patients were treated with EA for Tiaozang Xingshen at Baihui (GV 20) and Shenting (GV 24), and bilateral Feishu (BL 13), Pishu (BL 20), Shenshu (BL 23), Zusanli (ST 36), Sanyinjiao (SP 6), Hegu (LI 4) and Taichong (LR 3). EA was operated with disperse-dense wave, 2 Hz/100 Hz in frequency and 0.1 mA to 1.0 mA in current intensity; 30 min each time, once daily. One course of EA consisted of 5 treatments, at the interval of 2 days and the intervention lasted 8 courses. Before treatment in the control group, before and after treatment in the observation group, the score of Montreal cognitive assessment scale (MoCA), the score of clinical dementia rating (CDR), Flanker paradigm, Stroop paradigm, Nback paradigm, the score of self-rating anxiety scale (SAS), the score of self-rating depression scale (SDS), and the score of Hamilton depression rating scale (HAMD) were evaluated separately; the glycolipid metabolic indexes (fasting plasma glucose [FPG], glycosylated hemoglobin type A1c [HbA1c], total cholesterol [TC], triacylglycerol [TG], high-density lipoprotein cholesterol [HDL-C] and low-density lipoprotein cholesterol [LDL-C]) were determined；with the magnetic resonance spectroscopy technique adopted, the metabolites in the basal ganglia area were detected. The correlation analysis was performed for the metabolite values with MoCA score, CDR score , Flanker paradigm, Stroop paradigm, and Nback paradigm. RESULTS: Before treatment, compared with the control group, in the observation group, MoCA score was lower (P<0.001), CDR score and the levels of FPG and HbA1c were higher (P<0.001); the reaction times of Flanker non-conflict, Flanker conflict, Stroop neutrality, Stroop congruence, Stroop conflict, and 1-back were prolonged (P<0.05, P<0.001), and the accuracy of Flanker conflict, Stroop conflict, and 1-back decreased (P<0.05, P<0.01); the ratio of N-acetyl aspartate (NAA) to creatine (Cr) in the left basal ganglia area was dropped (P<0.001), and that of myo-inositol (MI) to Cr in the right side increased (P<0.05). In the observation group after treatment, compared with the levels before treatment, MoCA score was higher (P<0.001), the scores of CDR, SAS and HAMD were reduced (P<0.01, P<0.05), the reaction times of Flanker conflict and Stroop conflict shortened (P<0.001, P<0.05), and the accuracy of Flanker conflict and 1-back increased (P<0.001, P<0.05); the ratio of NAA to Cr in the left basal ganglia area and that of the gamma-aminobutyric acid (GABA) to Cr in the right increased (P<0.05), that of MI to Cr in the right decreased (P<0.05). Before treatment, in the observation group, the ratio of MI to Cr in the right basal ganglia area was positively correlated with the reaction time of Stroop congruence (r=0.671, P=0.012) and this ratio was positively correlated with the reaction time of Stroop conflict (r=0.576, P=0.039). CONCLUSIONS Electroacupuncture for "adjusting zangfu function and rescuing the mind" improves the cognitive function of T2DACD patients, which may be related to the regulation of NAA, MI and GABA levels in the basal ganglia.
Humans ; Electroacupuncture ; Acupuncture Therapy ; Acupuncture Points ; Diabetes Mellitus, Type 2/therapy* ; Glycated Hemoglobin ; Cognitive Dysfunction/therapy* ; Cholesterol ; gamma-Aminobutyric Acid