1.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
2.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
3.Effects of agaric polysaccharides on antioxidant capacity,serum immune indexes and intestinal flora of calves
Qiru WANG ; Yu ZHANG ; Guojun JIANG ; Haiyan LIU ; Xin HE ; Xinchao SHI ; Xing-Hua ZHAO
Chinese Journal of Veterinary Science 2024;44(6):1290-1298
This experiment was conducted to investigate the effects of agaric polysaccharides on an-tioxidant capacity,immune function,and intestinal flora of calves.Twenty-four healthy Holstein calves of(30±3)days of age and with the similar body weight at(55.33±1.86)kg were selected and randomly divided into two groups:the control group(group C)and test group(group T)with 12 replicates in each group and one calves in each replicate.Group C was fed starter and milk repla-cer,and group T was fed starter and milk replacer with 10 g of agaric polysaccharides to each calve for 10 d.Serum antioxidant,immune indexes and intestinal flora were tested.The results showed as follows:compared with group C,the enzyme activities of SOD and GSH-Px in serum of calves in group T were significantly increased(P<0.05),and were increased by 29.09%and 15.35%,re-spectively;compared with group C,IgA,IL-2 and TNF-α were significantly increased in T group(P<0.05).Adding agaric polysaccharides significantly increased the relative abundance of Firmi-cutes(P<0.05)and decreased the relative abundance of Proteobacteria in feces of calf(P>0.05);the relative abundance of Lactobalillus and Faecalibacterium were increased(P<0.05);the relative abundance of Butyricoccus-pullicaecorum was increased(P<0.05).LEfSe analysis results showed that there were 11 marker species in group T,such as Firmicutes and Lactobacillus,and 9 marker species in group C,such as Proteobacteria.The results showed that agaric polysaccharides could improve the antioxidant capacity and immune function of calves,and also could improve the structure of intestinal flora.
4.Isolation and identification of feline calicivirus and preparation of its inactivated vaccine
Yanmei YANG ; Junnan KE ; Yu QI ; Honglin REN ; Guojun ZHANG ; Zengshan LIU ; Liheng ZHANG ; Zhaozhe WANG ; Xianfeng LIU
Chinese Journal of Veterinary Science 2024;44(9):1892-1897
A virus was successfully isolated from a sick cat exhibiting clinical signs such as oral mu-cosal ulceration,nasal mucosal redness,and increased nasal secretions utilizing F81 cells.Through a comprehensive analysis as such PCR amplication,sequencing,morphology,serology,and animal re-gression tests,the virus was identified as a feline calicivirus and named FCV-BJ,an inactivated vac-cine was developed from this isolated strain its safety and efficacy were assessed.The results re-vealed that the isolated FCV-BJ strain exhibited characteristic serological and morphological fea-tures consistent with caliciviruses.Furthermore,inoculation of cats with the FCV-BJ demonstrated the strain is highly virulent and the cats manifested the clinical signs of feline calicivirus infection.For the vaccination trial,domestic cats were immunized with inactivated fifth-generation virus cell culture at varying dilutions,followed by a booster immunization after 21 days.Fourteen days after the challenge with the virus,cats immunized with 107.0 TCID50/mL or higher remained largely healthy,while all cats in the control group developed clinical signs of FCV.These findings suggest that the inactivated vaccine derived from the FCV-BJ isolate exhibits strong immunogenicity and protective efficacy at a minimum immunization dose of 107.0 TCID50/mL.This strain holds promise as a candidate for vaccine production,providing a valuable reference and foundation for future re-search and development of feline calicivirus vaccines.
5.Lycium Barbarum Polysaccharide Inhibits the Malignant Progression of Gallbladder Cancer Cells by Promoting Cellular Ferroptosis
Guojun XIN ; Jiancheng WANG ; Yong YANG
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2024;53(4):452-457
Objective To investigate the effects of lycium barbarum polysaccharide(LBP)on the proliferation and metastasis of gallbladder cancer cells(NOZ and SGC-996)by regulating cellular ferroptosis.Methods NOZ and SGC-996 cells were cul-tured in vitro by treatment with different doses of LBP.The cells were divided into control group,20 μmol/L LBP group,40μmol/L LBP group,60 μmol/L LBP group,80 μmol/L LBP group,and LBP+Fe-1 groups.Cell activity was detected by MTT assay.Cell proliferation was detected by EdU assay.Cell invasion ability was detected by Transwell assay.Fe2+level was detec-ted by colorimetric assay,reactive oxygen species(ROS)and malondialdehyde(MDA)levels were determined by BODIPYT 581/591 C11 molecular probe.Western blot was used to detect glutathione peroxidase 4(GPX4),acyl-CoA Synthetase long-chain family member 4(ACSL4),β-catenin,and Wnt3A protein expression.Results Compared with the control group,cell viability,proliferation,and invasion were significantly decreased in the 40 μmol/L LBP,and 60 μmol/L LBP groups(all P<0.05),and Fe2+level,ROS,and MDA activities were significantly increased(all P<0.05),and GPX4,β-catenin,and Wnt3A protein ex-pressions were significantly reduced,and ACSL4 protein expression was significantly increased(all P<0.05).Compared with the LBP group,cell viability,proliferation,and invasion ability were significantly increased in the LBP+Fer-1 group(all P<0.05).Conclusion LBP inhibits the proliferation and metastasis of gallbladder cancer cells by inducing ferroptosis.
6.Effects of S1PR5 on cognitive behavior and inflammatory response in li-popolysaccharide-induced mice and its anti-inflammatory mechanism
Zijing REN ; Guojun WU ; Jingxian WANG ; Shengguang ZHANG ; Peiyang ZHOU
Chinese Journal of Pathophysiology 2024;40(10):1916-1925
AIM:To explore the impact of sphingosine 1-phosphate receptor 5(S1PR5)on lipopolysaccha-ride(LPS)-induced neuroinflammation and cognitive-behavioral impairments in mice,alongside the anti-inflammatory im-pacts on BV2 cells and associated mechanisms.METHODS:(1)C57BL/6 wild-type(WT)mice and homozygous S1PR5 knockout(KO)mice were utilized and categorized into WT control,WT-LPS,S1PR5 KO control,and S1PR5 KO-LPS groups using the random number method.Neuroinflammatory models in mice were induced by a single intraperitoneal injection of 5 mg/kg LPS in the WT-LPS and S1PR5 KO-LPS groups,while an equivalent volume of saline was injected in-to the WT control and S1PR5 KO control groups.Following 7 days of modeling,the Morris water maze test was conducted,followed by the collection of brain tissues from each group of mice.Hippocampal tissue sections were stained with Nissl.The mRNA expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and IL-6 in hippocampal tis-sues were determined using RT-qPCR.Western blot and tissue immunofluorescence techniques were employed to assess the expression of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)in hippocampal tissues.(2)The BV2 cells underwent LPS stimulation to induce an inflammatory response and were treated with either the S1PR5 ago-nist A971432 or lentiviral overexpression of S1PR5.The effects of S1PR5 agonism or overexpression on S1PR5,IL-1β,IL-6,TNF-α,and CD206 were assessed using RT-qPCR.Additionally,CD206 expression was examined via cellular im-munofluorescence.Western blot was employed to analyze the protein levels of microglia polarization markers CD206,in-ducible nitric oxide synthase(iNOS),cyclooxygenase 2(COX2),and NLRP3,as well as p-NF-κB,cleaved caspase-1,and IκBα.RESULTS:(1)Findings from in vivo experiments indicated that S1PR5 KO notably exacerbated LPS-induced memory impairments in mice,alongside increased mRNA levels of IL-1β and IL-6,and increased protein levels of NLRP3 in the hippocampus.(2)The presence of S1PR5 in BV2 cells remained unaffected by variations in A971432 concentration and exposure duration.(3)Activation of S1PR5 or its overexpression significantly mitigated LPS-induced expression of IL-1β,IL-6,and TNF-α,while concurrently enhancing CD206 expression in BV2 cells at the mRNA level.At the protein level,it led to a noteworthy increase in CD206 expression,indicative of M2-type macrophages,and a reduction in the ex-pression of iNOS and COX2,markers of M1-type macrophages.Furthermore,it downregulated NLRP3,p-NF-κB,and cleaved caspase-1 expression,while upregulating IκBα expression.CONCLUSION:S1PR5 deficiency exacerbates cog-nitive deficits in mice by promoting neuroinflammatory responses induced by LPS.
7.Progress of High-Resolution Magnetic Resonance Vessel Wall Imaging in the Evaluation of Pathogenesis and Treatment of Intracranial Atherosclerosis-Related Ischemic Stroke
Xuelian WANG ; Fei WANG ; Xincheng LIU ; Tao YUAN ; Lijuan GAO ; Guojun TAN ; Guanmin QUAN
Chinese Journal of Medical Imaging 2024;32(7):747-752
Intracranial atherosclerosis has been regard as the most common cause of ischemic stroke.The information of the types and degree of vascular wall lesions is important basis for the diagnosis and treatment of ischemic stroke.High-resolution MR vessel wall imaging can directly display the lumen and wall of intracranial vessels,evaluate the characteristics of atherosclerotic plaque qualitatively and quantitatively,including intra-plaque hemorrhage,plaque enhancement,and plaque distribution.Thus,high-resolution MR vessel wall imaging may play an important role in risk stratification,pathogenesis,treatment and prognosis evaluation in ischemic stroke patients.This review summarizes the progress of high-resolution MRI vessel wall imaging in the evaluation of intracranial atherosclerosis,especially the pathogenesis of plaque features and treatment-related information.
8.Application of Thoracoscopy Combined With Laparoscopy for Radical Esophagectomy Based on Membrane Anatomy Theory
Jingtao WANG ; Bulang GAO ; Guojun WANG
Chinese Journal of Minimally Invasive Surgery 2024;24(1):1-6
Objective To investigate the feasibility and clinical significance of membrane anatomy theory in the application of thoracoscopic and laparoscopic radical esophageal resection.Methods A retrospective analysis was performed on 142 cases of thoracoscopic and laparoscopic radical esophagectomy based on membrane anatomy theory from December 2018 to October 2021.The esophageal mesangium,esophageal cancer,and nerves,blood vessels,lymphatic system,adipose tissue,upper stomach,left mesangium,and left gastric lymph nodes in the esophageal mesangium were removed as a whole.During the surgery,the space containing loose connective tissue around the esophagus was seen to be the esophageal fascial fusion space.The first 10 cases were labeled with nanocarbon tracer markers,showing esophageal lymphatic drainage to the left gastric lymph node.Results All the 142 patients had smooth surgery.The operation time was 150-230 min(mean,184.6±21.3 min),the intraoperative blood loss was 20-100 ml(mean,46.7±16.8 ml),the number of lymph nodes dissected was 12-41(mean,23.5±7.3),and the positive lymph nodes were found in 97 cases.The postoperative chest drainage time was3-10 d(mean,7.1±2.5 d),the postoperative oral intake time was 5-10 d(mean,7.6±1.7 d),and the total hospital stay was 9-20 d(mean,14.0±4.6 d).The total incidence of postoperative complications was 21.8%(31/142),including 7 cases(4.9%)of anastomotic leakage,9 cases(6.3%)of anastomotic stenosis,9 cases(6.3%)of hoarseness,and6 cases(4.2%)of residual gastritis.There was no postoperative bleeding,chyllevial leakage,infection,or death within 30 d after surgery.The follow-ups of the 142 patients lasted for 11-35 months,with a median of 26 months,and there was no recurrence and death.Conclusions There is a mesangial structure that constitutes an"envelope"around the esophagus.The membrane anatomy theory is suitable for the treatment of esophageal cancer,and radical resection of esophageal cancer based on the theory is safe,effective,and feasible.
9.Acupuncture with Manipulation for Lumbar Disc Herniation in Remission of 50 Cases:A Randomized Controlled Trial
Chun CHEN ; Jiao JIN ; Jingxuan MO ; Hai LIN ; Fudong SHI ; Guojun WANG ; Guannan WU ; Shimin ZHANG
Journal of Traditional Chinese Medicine 2024;65(10):1026-1032
ObjectiveTo evaluate the clinical effectiveness and safety of acupuncture with manipulation for lumbar disc herniation in remission period. MethodsOne hundred and four patients with lumbar disc herniation in remission were randomly divided into a treatment group and a control group, with 52 cases in each group. Treatment group applied acupuncture with manipulation of pointing, pulling, and shaking. Acupoints were selected as lumbar Jiaji (EX-B2, bilateral), Ashi point, Shenshu (BL 23, bilateral), Huantiao (GB 30, bilateral), Weizhong (BL 40, opposite side of the affected area), Chengshan (BL 57, opposite side of the affected area). The control group applied lumbar traction plus acupoint ultrasonic pulse penetration therapy (acupoints selection same as the treatment group); 20 minutes each time, 3 times a week, a total of 3 weeks for both groups. The primary outcome was the improvement rate of lumbar disc herniation symptoms and signs, which was calculated at 1 week of treatment, 3 weeks of treatment, 1 month follow-up, and 3 months follow-up, respectively; the secondary outcome were the Japanese Orthopaedic Association (JOA) scores, Visual Analogue Scale (VAS) scores, and Oswestry Disability Index (ODI) scores (including ODI total scores, sitting scores and standing scores), which were evaluated before treatment, 1 week of treatment, 3 weeks of treatment, 1 month follow-up, and 3 months follow-up; clinical effectiveness was assessed at 3 months follow-up; and the occurrence of adverse events in the participants, as well as blood routine, urine routine, stool routine, and electrocardiograms before and after the treatment were recorded to evaluate safety. ResultsTwo patients from each group fell out, and 50 patients of each group were included in the outcome analysis ultimately. The scores of lumbar disc herniation symptoms and signs improved more in the treatment group than in the control group at 1 week of treatment, 3 weeks of treatment, 1 month follow-up and 3 months follow-up (P<0.01). The JOA scores of participants in both groups at 1 week of treatment, 3 weeks of treatment, 1 month follow-up and 3 months follow-up were higher than those before treatment in the same group, and the VAS scores, ODI total scores, ODI sitting scores and standing scores were significantly lower than those before treatment in the same group (P<0.05), and the JOA scores of patients in the treatment group were higher than those of the control group at all time points, and the VAS scores, ODI total scores, ODI sitting scores and standing score were lower than those of the control group (P<0.05). At the 3 months follow-up, the excellent rate of the treatment group was 70.00% (35/50) better than that of the control group, which was 50.00% (25/50) (P<0.05). There were no abnormalities in blood, urine, stool routines and electrocardiograms before and after treatment in both groups, and no adverse events occurred. ConclusionAcupuncture with manipulation of pointing, pulling, and shaking for treating patients with lumbar disc herniation in remission has a better safety on pain relief and improving quality of life, and the effectiveness is better than lumbar traction plus acupoint ultrasonic pulse penetration therapy.
10.Clinical Efficacy of Medication of Xibining Ⅱ Prescription in Treatment of Knee Osteoarthritis with Cold-dampness Blockage Syndrome: A Retrospective Cohort Study
Zifeng CAO ; Wei MEI ; Li ZHANG ; Wendi PAN ; Xiaochen LI ; Jun MAO ; Guojun LI ; Peimin WANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(14):122-129
ObjectiveTo retrospectively analyze the clinical efficacy of Xibining Ⅱ prescription in the treatment of knee osteoarthritis with cold-dampness blockage syndrome by oral medication and to explore the influencing factors of endpoint events. MethodA real-world retrospective cohort design was adopted, and medical records of knee osteoarthritis patients with cold-dampness blockage syndrome treated with oral medication from the orthopedics outpatient department of Jiangsu Province Hospital of Chinese Medicine were collected. All patients received conventional Western medicine treatment and were divided into non-exposure group (573 cases) and exposure group (427 cases) according to whether or not they received treatment with Xibining Ⅱ prescription. Descriptive analysis of the baseline data of the 1 000 screened cases was performed using IBM SPSS 27.0. According to the baseline data, 334 pairs were matched using the propensity score matching method, resulting in a total of 668 cases in both groups. The changes in visual analogous scale (VAS), Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) total score, Japanese Knee Osteoarthritis Measure (JKOM) score, and traditional Chinese medicine (TCM) syndrome score before treatment and at 2, 6, 12 weeks after treatment, as well as the incidence of adverse reactions, were compared between the two groups. A multivariate logistic regression analysis was conducted to analyze the influencing factors of endpoint events, with clinical cure judged based on the improvement rate of WOMAC total score before and after treatment. ResultAfter 12 weeks of treatment, compared to the results before treatment, the VAS, WOMAC total score, JKOM score, and TCM syndrome score of patients in both groups significantly decreased (P<0.01). Compared to the non-exposure group, the exposure group showed a more significant reduction in VAS, WOMAC total score, JKOM score, and TCM syndrome score (P<0.01). After 12 weeks of treatment, the clinical cure rate and significant efficiency were higher in the exposure group than in the non-exposure group (P<0.05). Compared to the results before treatment within each group, VAS, WOMAC pain, stiffness, function scores, JKOM score, and TCM syndrome score significantly decreased at 2, 6, 12 weeks after treatment in both groups (P<0.01). Compared to the non-exposure group at the same time points, the exposure group showed a reduction in VAS at 2, 12 weeks, WOMAC pain at 6, 12 weeks, and function scores at 12 weeks (P<0.05, P<0.01). The JKOM score decreased at 6, 12 weeks, and the TCM syndrome score significantly decreased at 2, 6, 12 weeks in the exposure group (P<0.05, P<0.01). Multivariate logistic regression analysis at 12 weeks showed that factors affecting clinical cure included the course of disease, history of alcohol consumption, hypertension, coronary heart disease, and the use of Xibining Ⅱ prescription (P<0.05, P<0.01). Compared to the non-exposure group at the same time points, the incidence of epigastric discomfort in the exposure group was lower at 2, 12 weeks (P<0.01), the incidence of diarrhea and vomiting was slightly higher than that in the non-exposure group, but the difference was not statistically significant. ConclusionThe clinical application of Xibining Ⅱ prescription combined with conventional Western medicine treatment in the treatment of knee osteoarthritis with cold-dampness blockage syndrome is more effective than conventional Western medicine treatment alone. It can significantly reduce VAS, WOMAC total score, JKOM score, and TCM syndrome score, with more pronounced long-term effects and a low incidence of adverse reactions.

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