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 study the effects of T-2 toxin on expression of fibroblast growth factor 8 (FGF8) and fibroblast growth factor receptor 3 (FGFR3) in articular cartilage and subchondral marrow of rats under low selenium condition, and to explore the mechanism of deep cartilage injury and secondary complications in Kaschin-Beck disease (KBD).Methods:Twenty-four healthy male SD rats weighted 60 - 80 g were selected, they were divided into conventional feed group (selenium content of 101.5 μg/kg) and low-selenium feed group (selenium content of 1.1 μg/kg) by random number table method, with 12 rats in each group. After 30 days of feeding, the conventional feed group was further divided into control group and T-2 toxin group (100 μg·kg -1·d -1), and the low-selenium feed group was further divided into low-selenium group and low-selenium+ T-2 toxin group, with 6 rats in each group. After 30 days of feeding, the rats were sacrificed and the knee cartilage with cancellous bone was taken. Pathological changes of knee cartilage were observed by HE staining. Immunohistochemical method was used to detect the expression of FGF8 and FGFR3 in cartilage and subchondral marrow of knee joint, positive expression rates of FGF8 and FGFR3 in articular cartilage were calculated, and the integrated optical density (IOD) values of FGF8 and FGFR3 positive expression in subchondral marrow were analyzed by Image-Pro Plus 6.0 software. Results:Under light microscope, chondrocytes in low-selenium+ T-2 toxin group were sparse, and empty chondrocytes in the deep and middle layers of articular cartilage increased, and chondrocytes died and became red cell shadows. The extracellular matrix dissolved and was slightly stained in deep region, turning into necrotic and unstructurized areas. Proliferating granulation tissue was visible nearby. The positive expression rate of FGF8 in articular cartilage of rats in low-selenium+ T-2 toxin group [(88.61 ± 10.97)%] was higher than that in control, low-selenium and T-2 toxin groups [(10.35 ± 2.48)%, (19.26 ± 3.08)%, (58.89 ± 9.29)%, P < 0.05]; IOD value of FGF8 positive expression in subchondral marrow [(16.73 ± 1.72) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(1.20 ± 0.41) × 10 6, (4.33 ± 0.97) × 10 6, (12.80 ± 1.12) × 10 6, P < 0.05]. The positive expression rate of FGFR3 in articular cartilage of rats in low-selenium+ T-2 toxin group [(89.76 ± 8.59)%] was higher than that in control, low-selenium and T-2 toxin groups [(13.18 ± 2.25)%, (21.15 ± 2.33)%, (32.55 ± 6.72)%, P < 0.05]; IOD value of FGFR3 positive expression in subchondral marrow [(16.50 ± 5.36) × 10 6] was higher than that in control, low-selenium and T-2 toxin groups [(7.58 ± 1.02) × 10 6, (10.73 ± 7.13) × 10 6, (9.83 ± 5.63) × 10 6, P < 0.05]. Conclusion:Under low selenium condition, T-2 toxin changes expression of FGF8 and FGFR3 in deep chondrocytes of articular cartilage and subchondral marrow in rats, elevated expression of FGF8 and FGFR3 may be involved in the occurrence and development of secondary changes in KBD.

Objectives To identify and characterize 10 strains of Francisella philomiragia-like organisms isolated from blood samples and environmental water.Methods The 10 clinical and environmental isolates were identified by traditional morphological examination and biochemical characterization,matrix-assisted laser desorption/ionization time of flight(MALDI-TOF) mass spectrometry(MS) systems and sequencing based on 16S rRNA gene.The minimum inhibitory concentrations were tested by E-test methods.Results All the 10 isolates were gram-negative coccobacilli appearing tiny and faint counterstain of safranin,negative for urease,nitrate reduction and X and/or V factor requirement,but positive for oxidase and catalase.The isolates grew rapidly in sheep blood agar,chocolate agar and BCYE plate forming white opaque,colorless transparent or gray smooth colonies with about 2-mm diameters,but did not grow in M-H agar and MacConkey agar.The sequencing for 16S rRNA gene indicated that the 10 isolates shared more than 99.6％ similarity to Francisella philomiragia,and fell into the same clusters of Francisella philomiragia on phylogenetic tree.The MALDI-TOF MS analysis also showed the typical peaks with 6 153 m/z,5 180 m/z,7 757 m/z and 9 392 m/z which were similar to Francisella philomiragia ATCC 25015.However,they may be misidentified to be Sphingomonas paucimobilis by using Vitek 2 GN cards,Neisseria cinerea by using Vitek 2 NH cards,Myroides odoratimimus by using API 20NE strips and Haemophilus by using API NH cards.The results of antimicrobial susceptibility showed that they were all sensitive to chloramphenicol,doxycycline,tetracycline,gentamicin,ofloxacin and ciprofloxacin.Conclusion The 10 isolates could be identified as Francisella philomiragia,so we should pay more attention to the infrequent pathogen for its inactive biochemical reaction and the misidentification by commercial detection systems.

Objective To establish a mouse model of genital human papillomavirus (HPV) pseudovirion (PsV) transmission and evaluate the protective potency of HPV16 VLP vaccine.Methods HPV16 PsV with the encapsidated luciferase expressing plasmid Luc were generated from 293FT cells and purified by size-exclusion chromatography.The endpoint titers of HPV16 PsV-Luc were determined on 293FT cells, denoted as TRLU/mL.For in vivo genital challenge, mice were synchronized in a diestrus-like status by a subcutaneous injection with 0.1 μg β-estradiol and then with 3mg DepoProvera after 24 hours.Six hours prior to HPV16 PsV-Luc challenge, deeply anesthetized mice were intravaginally pretreated with 50 μL of 4%nonoxynol-9 ( N-9 ).HPV16 PsV-Luc was thoroughly mixed with 20 μL solution containing 4%carboxymethylcellulose ( CMC ) and intravaginally instilled using a positive-displacement pipette.Forty-eight hours after PsV-Luc challenge, mice were anesthetized and D luciferin was intravaginally instilled.After 3 minites, bioluminescence was measured with a cryogenically cooled Xenogen IVIS camera system.Then,the murine genital challenge model was used to determine the potency that HPV16 VLP vaccine is efficient at preventing HPV infection.Results HPV16 PsV-Luc was generated and purified from 293FT cells.HPV16 PsV-Luc was verified to containe L1 and L2 protein by Western blot.HPV 16 PsV-Luc successfully infected vaginal epithelial cells of mouse and the murine genital challenge model was established.To achieve consistent bioluminescence, the minimal dose of HPV16 PsV-Luc was 1.7 ×104 TRLU.The protective potency of HPV16 VLP vaccine was shown using this murine model.Our data showed that immune serum with over neutralizing antibody titer of 256-fold was sufficient to confer protection against HPV PsV genital infection .Conclusion The murine genital challenge model of HPV16 was successfully established, and the model is used to evaluate the potency of HPV16 VLP vaccine against in vivo genital HPV16 PsV challenge.Our model will benefit for the investigation of HPV neutralization and the potency evaluation of the HPV vaccine .

Objective:To observe the effects of different doses of advanced glycosylation end products ( AGEs ) on bFGF expression of cultured rabbit M üller cells in vitro.Methods:Immunocytochemistry and transmission electron microscopy methods were used identified cultured M üller cell.Immunocytochemistry method was used to semi-quantitate bFGF expression of retinal Müller cells at 640 μl/2 000 μl AGEs conditions.We observed effects of AGEs and PKC inhibitor Calphstion C on bFGF mRNA expression .Results:640 μl/2 000 μl AGEs stimulate bFGF expression of retinal Müller cell.Calphostin C inhibits bFGF mRNA increase stimulated by AGEs,and inhibition achieves strongest at concentration 50 nmol/L.Conclusion:AGEs can stimulate bFGF expression of Müller cell to exert the role of angiogenesis .bFGF mRNA expression may be regulated by activation of PKC pathway .

Objective To investigate the therapeutic effect of traditional Chinese medicine(TCM)recipe of nourishing kidney and activating blood for treatment of cisplatin induced acute kidney injury(AKI)in mouse model. Methods Twenty-one male BALB/c mice were randomly divided into three groups:control group,AKI model group and treatment group with the above TCM recipe(each n=7). The AKI model was reproduced by a single intraperitoneal injection of cisplatin 20 mg/kg;the TCM recipe with dosage of 10μL · g-1 · d-1 was given to the treatment group,while equal volume of normal saline was given to the model and control groups by gavage,lasting for 6 days in all groups. The changes of each mouse body weight were observed;mouse venous blood serum creatinine(SCr)level was detected,changes of renal tissue weight and its histopathology were observed under light microscope,and the score of acute tubular necrosis(ATN)was calculated. Results Compared to the control group,the body and renal weight in AKI model group were significantly lowered〔body weight(g):17.18±0.29 vs. 19.33±1.43,renal weight(g):0.28±0.01 vs. 0.32±0.11,both P<0.01〕,and SCr and ATN scores were significantly increased in AKI model group〔SCr(μmol/L):86.77±10.97 vs. 14.37±0.81,ATN score:3.33±0.52 vs. 0.17±0.41,both P<0.01〕. Compared to AKI model group,the body weight(g:18.70±0.28)and renal weight(g:0.31±0.01)in the treatment group were markedly increased,and SCr(μmol/L:21.98±5.52)and ATN scores(2.00±0.63)were significantly lower than those of the AKI model group(all P<0.01). Under optical microscope,there were mouse renal tubular epithelial cell edema and necrosis,renal interstitial inflammatory cell infiltration,the glomerular form roughly normal in the AKI model group;compared with the AKI model group,in the treatment group the degree of mouse renal tubular necrosis was significantly reduced and glomerular form was basically normal. Conclusion The TCM recipe of nourishing kidney and activating blood can reduce SCr and improve the pathological changes of renal tissue in cisplatin induced AKI mouse models,thus it has therapeutic effect for treatment of AKI in mice.

Objective To explore the effect of schisandra chinensis fruit ethanol extract on nephrin and desmin ex-pression in adriamycin(ADR) induced podocyte injury in vitro. Methods Conditionally immortalized mouse podocytes were treated with ADR for 24 h in vitro, then the medium was changed to medium with SE(250 mg/L)for 24 h. Podocytes were di-vided into four groups:control group,model group, SE intervention group and SE group. The expression of nephrin in podo-cytes was detected by immunofluorescence. Western Blot was employed to assess nephrin and desmin expression. Transcrip-tion level of nephrin and desmin were determined by qRT-PCR. Results Nephrin expression was distributed along the cell membrane in linear or granular pattern in control group and SE group. Fluorescence intensity in model group was lower than that of control group SE group and SE intervention group. There was no significant difference of nephrin and desmin protein and mRNA level between control group and SE group. Compared with the model group, protein and mRNA level of nephrin was lower than that of control group and SE intervention group. The protein expression and mRNA transcription of desmin in model group was higher than those in control group and SE intervention group (P＜0.05). Conclusion SE(250 mg/L)has no harmful effect on the podocytes in vitro. SE can protect the podocytes from damage by adriamycin in vitro. SE not only up-regulate the expression of nephrin, but also down-regulate of desmin expression.

Objective To study the change law between deficiency of kidney-Yang and stagnation of blood, and supply a study evidence for the therapy method of invigorate the kidney and promoting blood flow. Method 37 Wistar rats which were made of the model of deficiency of kidney-Yang, were randomly divided into three groups:control group, model group and treatment group. Changes including appearance, general objective sign, organ weight, microcirculation and blood rheology were recorded. Result Phenomenons, including rare fur, depression, lazy, duck stools and purpl dim in micrangium micrangium, occurred in model group. The weight of adrenal body and seminal vesicle and the contents of serum andrusol in model group were lower than those in control group. And in model group, significant increasing TC and LDL-C and decreasing HCL-C were observed. Conclusion Deficiency of kidney-Yang is associated with stagnation of blood, and the time delay of stagnation of blood can lead deficiency of kidney-Yang.

Objective:To investigate the expression of mmp-2 in mesangial proliferative glomerulonephritis(MsPGN) rat renal tissue and the effects of nourishing kidney and activating blood flow recipe of TCM. Methods: 54 Male SD rats were divided into control group, MsPGN group and treatment group with nourishing kidney and activating blood flow recipe of TCM. Immunohistochemical staining, flow cytometry, western blot and RT-PCR were used to check the protein expression of mmp-2. Results: In MsPGN group, following with the disease progressing, the glomerulus grew graduatly, ECM increased, the basement membrane of glomerulus was thicker. The immunohistochemical staining showed that the chief positive granules were deposited in glomerulus and renal tubular, the expression of mmp-2 was lower than that in control group, but in treatment group the expression of mmp-2 was higher than that in MsPGN group. Conclusion: In MsPGN renal tissue, the expression of mmp-2 was reduced, its activity was weakened. Nourishing kidney and activating blood flow recipe of TCM could induce the expression of MMP-2 and partly recover the activity. Nourishing kidney and activating blood flow recipe of TCM could resist glomerular sclerosis and postpone the course of chronic renal failure.

Objective:To investigate the possible interaction of Lp(a), and Lp(a) circulating immune complexes (CIC) 〔Lp(a)-CIC〕 level with triglyceride (TG). Methods:Plasma Lp(a), Lp(a)-CIC and LDL-CIC levels were determined by ELISAs in 232 patients with various dyslipidaemias, respectively. Results:Hypertriglyceridaemic patients exhibited the lowest plasma Lp(a) levels, while hypercholesterolaemic patients exhibited the highest. Patients with mixed hyperlipidaemia had intermediate serum Lp(a) concentrations, which were significantly lower than those in the controls. Interestingly, we also found that hypertriglyceridaemic patients exhibited the lowest plasma Lp(a)-CIC and LDL-CIC levels, while hypercholesterolaemic patients exhibited the highest. TG levels were negatively correlated with Lp(a) (r=-0.15, P