1.The role of bile acid dynamics in inflammatory bowel disease and metabolic dysfunction-associated steatotic liver disease
Si-Rui Yu ; Run Sun ; Mei-Ya Shi ; Mei Yang ; Kun Chen ; Qiong Pan ; Ling Zhao ; Ming-Yue Wu ; Jin Chai
Liver Research 2026;10(1):35-50
The prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) is higher among individuals with inflammatory bowel disease (IBD) than in the general population. Emerging evidence indicates that the development of MASLD in patients with IBD may occur independently of traditional metabolic risk factors, suggesting a unique pathophysiological mechanism distinct from conventional MASLD pathways. Bile acids (BAs), which act as critical signaling molecules in enterohepatic circulation, play an essential role in maintaining gastrointestinal homeostasis through bidirectional gut–liver axis communication. These molecules are increasingly recognized as pivotal regulators in the progression of both MASLD and IBD. While previous studies have characterized the dynamics of BA in blood or fecal samples under both conditions, a comprehensive understanding of their metabolic profiles and the associated interactions within the gut–liver axis is still lacking. This review synthesizes current evidence from studies employing BA metabolomics to investigate IBD and MASLD. By focusing on BA signatures, we summarize conserved alterations between these two conditions and their potential mechanisms of disease progression. Our review advances understanding of BA-mediated pathways in IBD and MASLD, providing a foundation for assessing their roles in contributing to the pathogenesis of MASLD in patients with IBD.
2.Effect of Folic Acid-modified Crebanine Polyethylene Glycol-polylactic Acid Hydroxyacetic Acid Copolymer Nanoparticles Combined with Ultrasonic Irradiation on Subcutaneous Tumor Growth of Liver Cancer in Mice
Rui PAN ; Junze TANG ; Hailiang ZHANG ; Kun YU ; Xiaoyu ZHAO ; Xin CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(4):217-225
ObjectiveTo investigate the effect of folic acid-modified crebanine polyethylene glycol-polylactic acid hydroxyacetic acid copolymer(PEG-PLGA) nanoparticles(FA-Cre@PEG-PLGA NPs, hereinafter referred to as NPs) combined with ultrasonic irradiation on subcutaneous tumor of liver cancer in Kunming(KM) mice. MethodsEighty-four healthy male KM mice were utilized to establish a subcutaneous tumor model of mouse hepatocellular carcinoma with H22 cells, then mice were randomly divided into model group, placebo group, hydroxycamptothecin group(8 mg∙kg-1), low, medium and high dose crebanine raw material groups(2, 2.5, 3 mg∙kg-1, hereinafter referred to as the low, medium and high dose crebanine groups, respectively), low, medium and high dose NPs groups(2, 2.5, 3 mg∙kg-1), and low, medium and high dose NPs combined with ultrasonic irradiation groups(2, 2.5, 3 mg∙kg-1, hereinafter referred to as the low, medium and high dose combination groups, respectively). The corresponding doses of drugs were administered via tail vein injection, the model group received no treatment, while the placebo group was injected with an equivalent amount of normal saline. Dosing was conducted for a total of 10 times on alternate days. The body mass of the mice was monitored, and parameters such as body mass change rate, thymus index, spleen index, tumor volume, tumor weight, relative tumor growth rate(T/C), and tumor inhibition rate(TGI) were calculated. Pathological changes in liver and kidney tissues as well as the tumor were observed by hematoxylin-eosin(HE) staining. Additionally, the levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), blood urea nitrogen(BUN) and creatinine(CREA) in serum of mice were detected by biochemical method. Furthermore, the effect of ultrasound on the distribution of NPs in subcutaneous tumors of mouse hepatocellular carcinoma was observed by in vivo imaging technique. ResultsAmong different treatment methods, the combination of NPs and ultrasound irradiation had the best therapeutic effect. Compared with the model group, the body mass growth rates of mice in the medium and high combination groups decreased, while the thymus index and spleen index increased, but there was no statistically significant difference in serum AST, ALT, BUN and CREA levels, indicating that NPs combined with ultrasound irradiation had little effect on the normal physiological state of the body, oth groups had TGI>40% and T/C<60%, indicating a clear anti-tumor effect. Pathological analysis showed that compared with the NPs groups, the combination groups exhibited varying degrees of necrosis in tumor cells, accompanied by less damage to the liver and kidneys. In vivo imaging of small animals showed that compared with the high dose NPs group, the high dose combination group had stronger tumor targeting ability(P<0.01). ConclusionNPs combined with ultrasonic irradiation can not only effectively targeted the drug to the tumor site, inhibit the subcutaneous tumor growth of mouse liver cancer, but also decrease damage to liver and kidney tissues.
3.Efficacy of Fufang Lingjiao Jiangya Pills with Different Proportions of Goat Horn Replacing Antelope Horn on Spontaneous Hypertensive Rats
Tengjian WANG ; Wanlu ZHAO ; Yang YU ; Yan LIU ; Kun CAO ; Zheyuan LIN ; Yue WU ; Lilan LUO ; Weizhi LAI ; Zhaohuan LOU ; Qiaoyan ZHANG ; Quanlong ZHANG ; Luping QIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):68-78
ObjectiveTo investigate the optimal ratio of goat horn replacing antelope horn in Fufang Lingjiao Jiangya pills and the blood pressure-lowering mechanism of this medicine. MethodsThe blood pressure-lowering efficacy of Fufang Lingjiao Jiangya pills with varying proportions of goat horn replacing antelope horn was evaluated on spontaneous hypertensive rats (SHR). In this experiment, 50 SHR rats were randomly grouped as follows: model (n=8), captopril (0.01 g·kg-1) (n=6), low-dose blank Fufang Lingjiao Jiangya pills (0.342 g·kg-1) (n=6), high-dose blank Fufang Lingjiao Jiangya pills (0.684 g·kg-1) (n=6), low-dose antelope horn-containing Fufang Lingjiao Jiangya pills (0.378 g·kg-1) (n=6), high-dose antelope horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1) (n=6), low-dose goat horn-containing Fufang Lingjiao Jiangya pills (0.378 g·kg-1) (n=6), and high-dose goat horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1) (n=6). Additionally, 8 WKY rats were used as the normal group. Drugs were administered by gavage for 4 weeks while an equal volume of distilled water was administered for the normal and model groups. Blood pressure was measured before administration, 3 h post administration, and biweekly thereafter. In the experiment for Fufang Lingjiao Jiangya pills with goat horn replacing antelope horn in different proportions, 48 SHR rats were randomly grouped as follows: model, blank Fufang Lingjiao Jiangya pills (0.684 g·kg-1), antelope horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1), 2× goat horn-containing Fufang Lingjiao Jiangya pills (0.824 g·kg-1), 4× goat horn Fufang Lingjiao Jiangya pills (0.969 g·kg-1), and 6× goat horn Fufang Lingjiao Jiangya pills (1.112 g·kg-1). The normal group included 8 WKY rats, and the normal group and model group received an equal volume of distilled water. The treatment lasted for 2 weeks, and blood pressure was recorded at various time points (pre-administration, 3 h post administration, and on days 4, 7, 10, and 14 of administration). Serum levels of angiotensin-converting enzyme (ACE), angiotensin Ⅱ(Ang Ⅱ), renin, and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. Histopathological changes in the heart, kidney, and thoracic aorta were observed by hematoxylin-eosin staining. The protein levels of ACE2, angiotensin Ⅱ type 1 receptor (AT1R), and angiotensinogen (AGT) in the kidney tissue were determined by Western blot, while the expression of nuclear factor (NF)-κB p65 and Toll-like receptor 4 (TLR4) in the thoracic aorta tissue was assessed by immunohistochemistry. ResultsCompared with the model group, all treatment groups showed lowered blood pressure (P<0.05, P<0.01), and the 6× goat horn-containing Fufang Lingjiao Jiangya pills group showed consistent blood pressure-lowering effect with the antelope horn-containing Fufang Lingjiao Jiangya pills group. Compared with the normal group, the model group showed elevated serum levels of ACE, Ang Ⅱ, renin, and IL-6, while the elevations were declined in the Fufang Lingjiao Jiangya pills groups (P<0.05, P<0.01). Pathological changes in the heart, kidney, and thoracic aorta were alleviated in all the treatment groups, with the 6× goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups exhibited the best effect. Western blot and immunohistochemistry results showed that all the treatment groups exhibited down-regulated protein levels of AT1R, AGT, NF-κB p65, and TLR4 and up-regulated protein levels of ACE2 (P<0.05, P<0.01) compared with model group, with the 6×goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups showcasing the best effect. ConclusionReplacing antelope horn with 6×goat horn in Fufang Lingjiao Jiangya pills can achieve consistent blood pressure-lowering effect with the original prescription. The prescription may exert the effect by inhibiting the renin-angiotensin-aldosterone system (RAAS) and TLR4/NF-κB signaling pathways.
4.Effects of preservation conditions on the survival numbers of seven Brucella strains preserved in 30%glycerol brain heart infusion medium
Yu FAN ; Hongyan ZHAO ; Qingqing XU ; Kun LI ; Guozhong TIAN ; Hai JIANG ; Lanyu LI ; Dongri PIAO
Chinese Journal of Zoonoses 2025;41(9):945-951
This study was aimed at comparing the effects of storage conditions on the Brucella survival number in brain heart infu-sion medium,to provide experimental data support for the selection of storage conditions for Brucella strains.We created bacterial sus-pensions of seven strains of Brucella melitensis,Brucella abortus,and Brucella suis(denoted A,B,C,D,E,F,G)by using 30%glycerol brain heart infusion medium,then conducted stability testing.For accelerated stability tests,the bacterial solution was placed in a 37℃chamber for 1,2,3,or 4 weeks continuously,and the Brucella survival number at each time point was measured.For freeze-thaw stability testing,the bacterial solutions were frozen at-80℃,transferred to room temperature for 4 hours,and melted in one freeze-thaw cycle,and the Brucella survival number was measured for 2,4,6,and 8 freeze-thaw times.For long-term stability testing,the bacterial solutions were stored at-80℃,and the Brucella survival number was measured at various time points during the first,third,sixth,and twelfth months of storage.A line graph was plotted to demonstrate the changes in Brucella survival number at each time point in the three stability tests.IBM SPSS Statistics 22 software was used to assess statistically significant differences in Brucella survival number at each time point and the initial Brucella survival number.Accelerated stability tests showed that the Bru-cella survival number of the seven strains significantly decreased,and a statistically significant difference in Brucella survival number was observed after 1 week.In freeze-thaw stability tests,with increasing freeze-thaw times,the Brucella survival number of the seven strains showed a decreasing trend.Strains C and F showed statistically significant differences with respect to the initial Brucella sur-vival number after two times of freeze-thaw;strains A and D showed statistically significant differences with respect to the initial Bru-cella survival number after four times of freeze-thaw;strain G showed statistically significant differences with respect to the initial Bru-cella survival number after six times of freeze-thaw;and strain E showed statistically significant differences with respect to the initial Brucella survival number after eight times of freeze-thaw.The data for strain B showed abnormalities.In long-term stability testing,the Brucella survival number for strains C and F after storage for 1 month showed a statistically significant difference with respect to the initial bacterial solution.No statistically significant difference in Brucella survival number was observed for strains B and E after stor-age for 1,3,6,or 12 months.The data for strains A,D,and G showed abnormalities.The sterile 30%glycerol brain heart infusion medium maintained the Brucella survival number in an-80℃environment,and freezing and thawing multiple times should be avoided.Compared with the traditional freeze-drying method,this method is not only easy to perform but also avoids potential bio-safety hazards,and provides a reliable Brucella culture preservation scheme for researchers in related fields.
5.Clinical application of the posterior to anterior malleolar extended lateral approach in the treatment of quadrimalleolar fractures
Yanjiang FENG ; Dawei ZHAO ; Guangrong YU ; Jiang XIA ; Kun ZHANG ; Zhen WANG
Chinese Journal of Orthopaedic Trauma 2025;27(1):46-51
Objective:To explore the clinical efficacy of the posterior to anterior malleolar extended lateral approach (PAMELA) in the treatment of quadrimalleolar fractures.Methods:A retrospective study was conducted to analyze the clinical data of 12 patients with quadrimalleolar fracture who had been admitted to Foot and Ankle Surgery Department, Henan Luoyang Orthopaedic & Traumatological Hospital from June 2022 to June 2023. There were 5 males and 7 females, with an age of (37.3±12.2) years and a duration from injury to surgery of (8.8±3.5) d. Open reduction and internal fixation of displaced Chaput tubercle fractures, lateral and posterior malleolar fractures were conducted through the PAMELA for all patients. The incision exposure, operation time, intraoperative bleeding, and incision healing were noted. Postoperatively, the fracture reduction was evaluated using the Burwell-Charnley criteria. The clinical efficacy was evaluated at the final follow-up using the ankle-hindfoot score of American Orthopaedic Foot & Ankle Society (AOFAS), visual analogue scale (VAS), and range of motion (ROM) of the ankle joint.Results:Good exposure of the Chaput tubercle, anterolateral tibiotalar joint, and lateral and posterior malleoli was achieved during surgery in all patients. The follow-up time for the 12 patients was (14.3±1.8) months, the operation time (152.5±26.0) minutes, and the intraoperative bleeding (137.5±44.1) mL. All incisions healed at the first stage postoperatively without any complications. According to the Burwell-Charnley criteria, anatomic reduction was achieved in all patients. CT scans showed good reduction of the distal tibiofibular syndesmosis. At the final follow-up, their AOFAS score was (94.1±8.3) points, VAS 0 (0, 1) point, and ankle joint ROM 17.5°±9.0° for dorsiflexion and 35.2°±9.6° for plantarflexion.Conclusions:In the treatment of quadrimalleolar fractures, because the PAMELA can lead to good exposure of the anterolateral ankle joint, distal tibiofibular syndesmosis, and lateral and posterior malleolar fractures, it results in a high rate of anatomic reduction of the fractures, safe incisions and limited soft tissue complications. Therefore, it is a safe, simple, and effective surgical approach.
6.Symptoms and quality of life benefits of successful percutaneous coronary intervention in left main disease and/or 3-vessel disease patients with diabetes
Bo-da ZHU ; Tian-tong YU ; Peng HAN ; Bo-hui ZHANG ; Xi ZHANG ; Ping YUAN ; Gang WANG ; Yi YANG ; Hui-li ZHU ; Pan-pan SUN ; Tong-tong LI ; Shuai ZHAO ; Cheng-xiang LI ; Kun LIAN
Chinese Journal of Interventional Cardiology 2025;33(2):93-100
Objective To investigate whether successful percutaneous coronary intervention(PCI)could improve symptoms and quality of life(QOL)in left main disease and/or 3-vessel disease patients with diabetes.Methods Patients with left main disease and/or 3-vessel disease who underwent PCI in the First Affiliated Hospital of Air Force Medical University from April 2018 to May 2021 were consecutively enrolled and subdivided into 2 groups:diabetes and no diabetes.Detailed baseline characteristics,symptoms,including dyspnea and angina,assessed with the Rose dyspnea scale(RDS),Seattle angina questionnaire(SAQ),the European quality of life-5 dimensions(EQ-5D)and 12-item short-form health survey(SF-12)questionnaire respectively,procedural details,and 1 month and 1 year follow-up data were collected.Results Among 440 left main disease and/or 3-vessel disease patients,disease was present in 176(40.00%),who had more hypertension,peripheral artery disease,and LCX lesion(all P<0.05).The incidence of major adverse cardiovascular events(MACE)and all-cause mortality were similar between the two groups(both P>0.05)at 1 month follow-up,while all-cause mortality in diabetes patients was significantly higher than those without diabetes at 1 year follow-up(P=0.013).Low left ventricular ejection fraction was an independent risk factor for MACE and all-cause mortality at 1 month and 1 year follow-up after successful revascularization(all P<0.05).Most importantly,symptoms,including dyspnea and angina,and QOL were markedly improved regardless of diabetes both at 1 month and 1 year follow-up(all P<0.05).Diabetes patients showed improved dyspnea and QOL at similar degree to the non-diabetes patients(all P>0.05)and a more significantly relieved angina(P=0.013).Additionally,the number of chronic total occlusion(CTO)per patient was identified as an independent risk factor of dyspnea(OR 0.723,95%CI 0.525~0.997,P=0.048)and angina relief(OR 0.686,95%CI 0.473~0.995,P=0.047),and the contrast volume(OR 0.995,95%CI 0.992~0.999,P=0.008)as an independent risk factor of QOL improvement in diabetic patients.Conclusions Successful PCI is beneficial for relieving symptoms and improving quality of life in patients with diabetes who have left main disease and/or 3-vessel disease.
7.Mechanism of adipose mesenchymal stem cell exosomes inhibiting atopic dermatitis
Jia-qi BI ; Zhao WANG ; Bing-kun WANG ; Chun-yan SUN ; Ya SUN ; Xiao-tong CUI ; Xin PANG ; Xiao-yu WANG ; Jie-qiong WANG
Chinese Pharmacological Bulletin 2025;41(6):1148-1157
Aim To study the mechanism of adipose mesenchymal stem cell exosomes(ASC-exo)inhibition of fluorescein isothiocyanate(FITC)-induced atopic dermatitis(AD).Methods The mouse age,extrac-tion method,and the concentration of a solution of typeⅠ collagen enzyme and other conditions were compared to study the effects on the morphology and quantity of adipose mesenchymal stem cells(ASCs)after extrac-ted.FITC-induced mouse model in vivo was estab-lished and different doses of ASC-exo were given to measure ear thickness,ear weight and ear scratching times of mice.HE staining was used to observe the pathological changes of ear tissue of mice.The non-toxicity of ASC-exo was detected.IgE,IL-5,IL-13 and other cytokines were detected by ELISA.The gene ex-pressions of TSLP,IL-33,occludin,Claudin-1(CLDN-1)and E-cadherin were detected by RT-qPCR.The protein expression was detected by immunohistochemis-try.Results An efficient method for extracting ASCs was established.Compared with the blank group,mice in the model group showed obvious AD symptoms.Compared with the model group,ASC-exo administra-tion group significantly reduced the number of ear scratches,epidermal thickening,inflammatory cell infil-tration and the secretion of Th2 cytokines IL-5 and IL-13.Meanwhile,ASC-exo administration group signifi-cantly increased the expression of structural proteins CLDN-1 and occludin in epithelial cells and decreased the expression of TSLP and IL-33.Conclusions ASC-exo can significantly improve Th2 skin inflamma-tion in AD mice,and its mechanism may be through in-creasing the expression of tight junction proteins and adhesion link protein in epithelial cells,repairing the skin barrier,and inhibiting the key promoters of allergy TSLP and IL-33.
8.Comparison of clinical manifestations,laboratory characteristics,and treatment outcomes of 258 patients with acute and chronic brucellosis
Xu ZHAO ; Ke-mei NIU ; Xia GAO ; Chun-xu SONG ; Yu FAN ; Qing-qing XU ; Zhong-rong LU ; Kun LI ; Feng GAO ; Mei-chun HAO ; Bing-zhi LIU ; Hai JIANG
Chinese Journal of Zoonoses 2025;41(6):660-667
To compare and analyze the clinical manifestations,laboratory characteristics,imaging findings,and treatment outcomes of patients with acute and chronic brucellosis,a retrospective analysis was conducted on 258 patients with brucellosis(202 in the acute group and 56 in the chronic group)hospitalized in Xinkang Hospital in Dalad Banner,Ordos City,Inner Mongolia Autonomous Region,from November 2023 to November 2024.General data,epidemiological characteristics,clinical presentations,laboratory test results,imaging findings,treatment outcomes,and prognosis were collected.The incidences of fever(51.5%vs 7.1%),fatigue(30.2%vs 12.5%),joint pain(42.9%vs 16.1%),and muscle pain(9.9%vs.1.8%)were significantly higher in the acute phase group(all P<0.05).The incidence of osteoarthritis complications was higher in the chronic brucellosis group(51.8%vs 8.9%,χ2=75.697,P<0.01).Univariate ANOVA analysisshowed that the Serum Agglutination Tests(SAT),alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBIL),creatinine(CRE),C-reactive protein(CRP),erythrocyte sedimentation rate(ESR),and bone destructionexhibited statistically significant differences between the acute and chronic phases of brucellosis(all P<0.05).Multivariate logistic regression analysis indicated that abnormal ALT(OR=14.18,95%CI:1.11-181.72;P=0.041)and bone destruction(OR=0.16,95%CI:0.04-0.63;P=0.009)were associated with chronic brucellosis.After treatment,all patients experienced have symptom relief in varying degrees,with 157 patients(60.9%)cured and 101 patients(39.1%)symptomatic improved(P<0.01).In conclusion,the incidences of fever,fatigue,and joint pain in patients during the acute phase is significantly higher than that those in patients during the chronic phase,while the incidence of osteoarthritis complications is higher in chronic phase patients.The incidences of abnormal SAT,ALT,AST,TBIL,CRE,CRP,and ESR,and bone destruction varies at different stages of brucellosis.Of those,abnormal ALT and bone destruction show a stronger association with,which can assist the clinical staging of brucellosis.
9.Application of Cytb and 12S rRNA in wildlife species identification for forensic science
Dezhi JIANG ; Yaxin ZHANG ; Yu ZANG ; Maolei AN ; Zan ZHANG ; Chengcheng QIU ; Yaoheng JIANG ; Wei SONG ; Hong ZHAO ; Kun XIE ; Jiayi CHEN ; Riga WU
Chinese Journal of Forensic Medicine 2025;40(3):308-311,322
Objective To analyze and compare the efficacy of DNA barcode,i.e.,Cytochrome b(Cytb)and 12S ribosomal RNA(12S rRNA)gene sequences,in the species identification of wildlife.Methods DNA extraction,quantification,PCR amplification of Cytb and 12S rRNA gene fragments,Sanger sequencing,and sequence alignment analysis were performed on ten wildlife samples.Results Both gene fragments were successfully amplified in six samples,while Cytb alone was successfully amplified in 1 sample,and 12S rRNA alone in 3 samples.Sequence analysis indicated that Cytb enabled species-level identification for 6 samples(Gallinula chloropus,Streptopelia orientalis,Phasianus colchicus,Falco naumanni,Myiopsitta monachus and Lynx lynx)and genus-level identification for 1 sample(Lepus).In contrast,12S rRNA achieved species-level identificaggion for 8 samples(Gallinula chloropus,Lepus sinensis,Phasianus colchicus,Myiopsitta monachus,Muntiacus reevesi,Macaca mulatta and Lynx lynx),representing seven species,and genus-level identification for 1 sample(Falco).However,by combining Cytb and 12S rRNA,all samples could be identified to the species level.Conclusion When applying DNA barcodes to wildlife identification,the Cytb and 12S rRNA gene regions analyzed here can effectively identify common species such as Gallinula chloropus and Streptopelia orientalis,but face difficulties in distinguishing closely related species within the same genus.Therefore,when conducting wildlife species identification,it is recommended to use two or more DNA barcode markers.
10.Effects of preservation conditions on the survival numbers of seven Brucella strains preserved in 30%glycerol brain heart infusion medium
Yu FAN ; Hongyan ZHAO ; Qingqing XU ; Kun LI ; Guozhong TIAN ; Hai JIANG ; Lanyu LI ; Dongri PIAO
Chinese Journal of Zoonoses 2025;41(9):945-951
This study was aimed at comparing the effects of storage conditions on the Brucella survival number in brain heart infu-sion medium,to provide experimental data support for the selection of storage conditions for Brucella strains.We created bacterial sus-pensions of seven strains of Brucella melitensis,Brucella abortus,and Brucella suis(denoted A,B,C,D,E,F,G)by using 30%glycerol brain heart infusion medium,then conducted stability testing.For accelerated stability tests,the bacterial solution was placed in a 37℃chamber for 1,2,3,or 4 weeks continuously,and the Brucella survival number at each time point was measured.For freeze-thaw stability testing,the bacterial solutions were frozen at-80℃,transferred to room temperature for 4 hours,and melted in one freeze-thaw cycle,and the Brucella survival number was measured for 2,4,6,and 8 freeze-thaw times.For long-term stability testing,the bacterial solutions were stored at-80℃,and the Brucella survival number was measured at various time points during the first,third,sixth,and twelfth months of storage.A line graph was plotted to demonstrate the changes in Brucella survival number at each time point in the three stability tests.IBM SPSS Statistics 22 software was used to assess statistically significant differences in Brucella survival number at each time point and the initial Brucella survival number.Accelerated stability tests showed that the Bru-cella survival number of the seven strains significantly decreased,and a statistically significant difference in Brucella survival number was observed after 1 week.In freeze-thaw stability tests,with increasing freeze-thaw times,the Brucella survival number of the seven strains showed a decreasing trend.Strains C and F showed statistically significant differences with respect to the initial Brucella sur-vival number after two times of freeze-thaw;strains A and D showed statistically significant differences with respect to the initial Bru-cella survival number after four times of freeze-thaw;strain G showed statistically significant differences with respect to the initial Bru-cella survival number after six times of freeze-thaw;and strain E showed statistically significant differences with respect to the initial Brucella survival number after eight times of freeze-thaw.The data for strain B showed abnormalities.In long-term stability testing,the Brucella survival number for strains C and F after storage for 1 month showed a statistically significant difference with respect to the initial bacterial solution.No statistically significant difference in Brucella survival number was observed for strains B and E after stor-age for 1,3,6,or 12 months.The data for strains A,D,and G showed abnormalities.The sterile 30%glycerol brain heart infusion medium maintained the Brucella survival number in an-80℃environment,and freezing and thawing multiple times should be avoided.Compared with the traditional freeze-drying method,this method is not only easy to perform but also avoids potential bio-safety hazards,and provides a reliable Brucella culture preservation scheme for researchers in related fields.


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