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".

Objective:To study the multi-locus sequence typing (MLST) gene characteristics of Brucella isolates in Guizhou Province. Methods:Brucella strains, which were isolated from 2017 to 2021 in Guizhou Province (preserved in the Bacterial and Viral Seed Bank of Guizhou Center for Disease Control and Prevention) were identified Brucella and species/types by BCSP31-PCR and AMOS-PCR methods, respectively. MLST method was used for genotyping, and Biometrics 8.0 software was used for cluster analysis of the typing results. Results:A total of 32 strains of Brucella were isolated in Guizhou Province and identified as Brucella melitensis ( B.melitensis) by BCSP31-PCR and AMOS-PCR methods. These strains were classified into 2 ST types (ST8 and ST39) by MLST method, with 28 strains of ST8 type(87.5%) and 4 strains of ST39 type (12.5%). The 28 strains of ST8 type were distributed in 7 cities (prefectures) of Guizhou Province, while the 4 strains of ST39 type were only found in Qianxinan Buyi and Miao Autonomous Prefecture. The cluster analysis results showed that ST8 and ST39 types strains were clustered in a group with the reference strain of B.melitensis, and there was only one nucleotide site difference between ST39 and ST8 types in the glk gene, indicating a close genetic relationship. Conclusions:B.melitensis is the main pathogen of the brucellosis epidemic in Guizhou Province in recent years. ST8 is the dominant MLST genotype in Guizhou Province.

Objective:To establish the quality evaluation method of Astmgali Radix based on three-dimensional fluorescence spectroscopy. Methods:The three-dimensional fluorescence spectra of Astmgali Radix extract was measured by fluorescence spectrum analysis technology, and the characteristics of fluorescence components of three-dimensional fluorescence spectrum of Astmgali Radix were analyzed by parallel factor analysis. The three-dimensional fluorescence spectra of Astmgali Radix samples from 23 different places were measured for quality discriminant analysis. Results:After the optimization, the three-dimensional fluorescence spectrum of 80% ethanol extract of Astmgali Radix showed that three groups of characteristic excitation/emission (λex/λem) peak, which located at 305 nm/420 nm (Peak 1), 280 nm/315 nm (Peak 2) and 265 nm/475 nm (Peak 3), respectively. The results of parallel factor analysis showed that Peak 1 and Peak 3 both contained one isoflavones fluorescent component, and Peak 2 contained two amino acid fluorescent components. There were differences in the number of characteristic peaks and fluorescence intensity of three-dimensional fluorescence spectra of Astmgali Radix samples from different origins. Conclusion:The three-dimensional fluorescence spectrum could evaluate the consistency of Astmgali Radix from different places. The method is simple, fast, and efficient. This method is accurate, which could provide reference for the quality evaluation of Astmgali Radix.

Objective: To investigate the effects of Shenqi Yizhi Granules on PI3K/AKT signaling pathway in Aβ1-42 bilateral hippocampal injection of AD model rats. Methods: A rat model of AD was established by bilateral hippocampus injection of Aβ1-42. Shenqi Yizhi granules were used for 60 days. Morris water maze was used to detect the learning and memory function of each rat. HE staining was used to observe the pathological changes of rat hippocampal CA3 area. Immunohistochemistry was used to detect the expression of PI3K and AKT protein in hippocampus. The relative expression levels of PI3K mRNA and AKT mRNA were detected by RT-PCR. Results: Compared with the blank group, the escape latency of the model group was significantly prolonged, and the number of entering the platform, the time and percentage of crossing the platform quadrant decreased significantly (P ＜ 0.05). At the same time, the vertebral cells in the hippocampal CA3 area were disordered and the neurons showed obvious lesions. PI3K/AKT showed significant inhibition at both protein and gene levels (P ＜ 0.05). After intervention with Shenqi Yizhi Granule, the escape latency of the model rats was significantly shortened, and the number of entering the platform and the time of crossing the platform quadrant were significantly increased (P ＜ 0.05). Conclusion: Shenqi Yizhi Granules can improve the cognitive function of Aβ1-42 bilateral hippocampus injection in AD rats, and its mechanism may be related to activation of PI3K/AKT signaling pathway in brain.

Objective: To investigate the effects of compatibility of Radix Ginseng and Radix Scutellariae (CRH) on learning and memory ability in multi-infarct dementia (MID) model rats and to explore the mechanism from the perspective of brain energy. Methods: We established MID rat model by thrombus injection, and learning and memory ability of MID rats was evaluated by Morris water maze; HE staining was used to observe the pathological changes in hippocampal CA1 region of rats; the contents of ATP, ADP and AMP in brain tissues were determined by highperformance liquid chromatography; flow cytometry was used to detect the changes of mitochondrial swelling and mitochondrial membrane potential in hippocampus of rats. Results: Compared with the rats in model group, the escape latency was significantly shortened in both high and low dose groups (P ＜ 0.05) . The times of crossing the platform were increased in both high and low dose groups (P ＜ 0.05) . The disorder of cell arrangement and loss of number of cell layers were significantly improved in high and low dose groups (P ＜ 0.05) . The degree of mitochondria swelling was significantly reduced in low dose group (P ＜ 0.05) . The decreasing trend of mitochondrial membrane potential was improved (P ＜0.05), and the energy charge of brain tissue was increased in high dose group rats (P ＜ 0.05) . Conclusion: CRH improved the cognitive decline and increased the concentration of energy charge in MID rats which was in relation to protect mitochondrial damage and improve brain energy, and further protect neurons.

Objective: To investigate the effects of EI injection on learning and memory ability and brain energy of two-way Meynert basal injection of Ibotenic acid (IBO) dementia model rats. Methods: A rat model of dementia wasestablished by bilateral meynert basal injection of IBO. After 8 weeks of EI injection, Morris water maze was used todetect the learning and memory ability of rats. Congo red staining was used to observe the deposition of Aβ plaque inhippocampal CA1 and cortical areas of rats. The changes of ATP, ADP and AMP in brain tissue of each group weredetermined by HPLC. The content of insulin in rat brain tissue was detected by ELISA kit. The expression of key proteinin PI3K/AKT signaling pathway was detected by Western blot. Results: Compared with the sham operation group, theescape latency of the model group was significantly prolonged, the number of entering the platform, the time andpercentage of crossing the platform quadrant decreased significantly (P ＜ 0.05); Aβ plaque deposition was observed inthe hippocampus and cortex; ATP/AMP ratio and insulin content were significantly decreased (P ＜ 0.05); brain tissue PI3 K and AKT protein were low expression (P＞ 0.05) . After intervention with EI injection, the escape latency of themodel rats was significantly shortened, the number of entering the platform and the time of crossing the platform quadrantincreased significantly (P ＜ 0.05); the hippocampus and cortex red staining was alleviated; the brain tissue ATP/AMPratio and insulin content increased significantly (P ＜ 0.05) . Conclusion: EI injection can improve the learning andmemory function of IBO-induced dementia model rats, which is related to the improvement of brain energy.

Objective To investigate the association between malnutrition parameters and serum N-terminal pro-brain natriuretic peptide (NT-proBNP) level in male patients aged ≥ 80 years with normal ejection fraction.Methods A total of 197 elderly male patients aged 80 95 years (average 85.4±9.7) were enrolled.Each patient received echocardiograms following admission,and ejection fraction was calculated.Serum concentrations of NT-proBNP were measured by electrochemiluminescence immunoassay.Multiple Logistic regression analyses and receiver operating characteristic (ROC) curves were used to assess the association between malnutrition parameters and serum level of NT-proBNP.Results Serum NT-proBNP level was higher in patients with malnutrition than in the controls [(711.9±1063.3) ng/L vs.(1375.2-±1891.7) ng/L,P=0.006].Pearson correlation analysis showed that the malnutrition parameters such as body mass index (BMI),blood hemoglobin,albumin and pre-albumin were negatively correlated with serum NT proBNP level (all P＜0.05).Logistic regression analysis showed that BMI and serum pre albumin level were independently associated with serum NT-proBNP level elevation (P=0.028 and 0.006,respectively).ROC curve analysis showed that the malnutrition parameters could predict the serum NT-proBNP level elevation.The area under the ROC curves for BMI,levels of blood hemoglobin,albumin and prealbumin in predicting serum NT proBNP level elevation was 0.7 (P=0.002),0.7 (P=0.004),0.6 (P=0.036),0.6 P=0.002),the sensitivity was 75.5％,51.0％,38.8％,89.8％,and the specificity was 54.4％,70.2 ％,84.2％ and 31.6 ％,respectively.Conclusions The malnutrition parameters including BMI,blood hemoglobin,albumin and pre-albumin can predict serum NT-proBNP level elevation.BMI and serum pre-albumin level are independently associated with serum NT-proBNP level elevation in male patients aged ≥ 80 years with normal ejection fraction.

OBJECTIVETo investigate clinical characteristics and changes of pulmonary imaging of mineral oil aspiration pneumonia in children.

METHODThe clinical features, CT findings, and effects of corticosteroid therapy were analyzed in 16 children with mineral oil aspiration pneumonia, who were hospitalized in our hospital from January 2003 to July 2013.

RESULTAll patients with mineral oil aspiration pneumonia had a history of mineral oil administration.Four patients had no clinical manifestations. Ten cases presented fever, and 8 of the 10 patients had fever in 4-8 h after taking mineral oil, and the temperature was between 39-40 °C. There were wheezing in 2 cases, shortness of breath in 6 cases, cyanosis in 1 case, dyspnea in 3 cases, and moaning in 2 cases, chest pain in 1 case, headache and abnormal EEG in 1 case.Six patients had rales in lungs. Peripheral blood white cells increased in 10 cases, and C- reactive protein elevated in 7 patients. Chest CT examination showed abnormal findings in 6 children, and the earliest CT was performed within 2 h after the accident. The rest 10 children got chest X-ray, and 9 of 10 children had abnormal findings. The earliest X-ray was done within 3 h after the accident. And the remaining 1 of 10 children showed no significant changes in the first chest X-ray 2-3 h after the accident until 3 days. All of the patients received corticosteroid and antibiotic treatments, 4 cases underwent bronchoalveolar lavage, 3 patients were given albumin, 6 cases received intravenous immunoglobulin. Three cases delayed in treatment with hormone because of misdiagnosis, and 2 of them had clearly secondary infections. Twelve patients recovered completely from oil aspiration pneumonia after 8 days to 5.5 months.

CONCLUSIONOil aspiration pneumonia in children occurs in almost all cases after mineral oil aspiration. Pulmonary opacities can be found by chest CT in most patients within 24 hours after mineral oil aspiration. Corticosteroids therapy was effective for patients with exogenous lipid pneumonia, which may inhibit the inflammatory response and possible pulmonary fibrosis.

Anti-Bacterial Agents ; therapeutic use ; C-Reactive Protein ; analysis ; Child, Preschool ; Diagnosis, Differential ; Female ; Fever ; diagnosis ; drug therapy ; Glucocorticoids ; therapeutic use ; Humans ; Infant ; Lung ; diagnostic imaging ; pathology ; Male ; Mineral Oil ; adverse effects ; Pneumonia, Lipid ; diagnosis ; drug therapy ; etiology ; Radiography, Thoracic ; Retrospective Studies ; Tomography, X-Ray Computed

Objective To observe the impact of coal-burning-borne fluorosis on the opening day of vagina,and estrogen cycle and endocrinology of female rat.Methods Ninety weaned two weeks and healthy female SD rats,weighing 60-80 g were selected,and divided into three groups by body weight using random number table:control group,moderate fluoride exposed group,and higher fluoride exposed group,thirty rates in each group.Rats in fluoride exposed groups were fed with baked corn collected from coal-burning-borne fluorosis areas containing different levels of fluorine(fluorine contents were 47.8 and 96.0 mg/kg,respectively) to establish coal-burning-borne fluorosis models.The corn fed to control group rats were collected from non-endemic areas (fluoride content was 5.2 mg/kg).After exposed to fluoride for 60,120 and 180 days,the rats were put to death during estrus in three batches (10 rats in every batche).The situation of female vaginal opening was observed and recorded every morning at 8:00 at the beginning of the experiment.Ten female rats in each group were randomly selected to observe vaginal exfoliated cells for 10 days continuously every morning,and HE staining was selected to discriminate estrogen cycle.Femoral artery blood was collected to observe the impact of estradiol(E2) in estrus.Results The vaginal opening day age of the longer fluoride exposed group [(49.0 ± 4.5)d] was longer than that of the control group[(45.4 ± 2.3)d] and the moderate fluoride exposed group[(46.7 ± 1.9)d,all P ＜ 0.05].Estrogen cycle of the longer fluoride exposed group[(5.8 ± 0.8)d] after fluoride exposure for 120 d was higher than that of the control group [(4.4 + 0.9)d] and the moderate fluoride exposed group [(4.9 ± 0.7)d,all P ＜ 0.05].Estrogen cycle increased with increasing dosage of fluoride after fluoride exposure for 180 d in three groups [control group:(4.3 ± 0.7)d,moderate fluoride exposed group:(5.4 ± 1.1)d,higher fluoride exposed group:(6.7 + 0.9)d,all P ＜ 0.05].Estrogen cycle in higher fluoride exposed group at 180 d[(6.7 ± 0.9)d] was higher than that of 60 d and 120 d[(5.2 ± 1.1),(5.8 ± 0.8)d,all P ＜ 0.05].At 60 d after exposed to fluoride,E2 levels of fluoride exposed groups were increased[control group:(30.125 ± 94.17)ng/L,moderate fluoride exposed group:(438.75 ± 71.80) ng/L],higher fluoride exposed group (456.25 ± 57.80)ng/L with increasing dosage of fluoride (all P ＜ 0.05).E2 level in rats exposed to fluoride for 180 d [(275.00 ± 127.83)ng/L] was lower than that of 60 d and 120 d [(456.25 ± 57.80),(392.50 ± 102.50)ng/L,all P ＜ 0.05] in higher fluoride exposed groups.Conclusions Coal-burning-borne fluorosis has a significant effect on vaginal opening day age,estrogen cycle and endocrinology of female rats.The higher the fluoride dose,and the longer the exposure time,the more severe the impact.