ObjectiveTo analyze the phylogroups of Klebsiella pneumoniae isolates from laboratory rats and mice housed at the Laboratory Animal Center Fudan University, optimize the identification method for Klebsiella pneumoniae, and perform virulence genes detection and antimicrobial susceptibility testing, thereby providing a practical basis for microbial detection strategies and prevention and control measures in laboratory animal facilities. MethodsBetween September 2023 and September 2025, ileocecal contents were collected from laboratory rats and mice purchased from laboratory animal suppliers, from which 11 strains of Klebsiella pneumoniae were isolated and cryopreserved. These strains were tested for β-lactamase genes using multiplex PCR for phylogrouping. Furthermore, representative strains from different phylogroups were selected for biochemical tests, mass spectrometry analysis, 16S rRNA gene and tyrB gene sequencing, as well as virulence genes detection and antimicrobial susceptibility testing. ResultsMultiplex PCR detection of β-lactamase genes revealed that among 11 strains, 4 were identified as Klebsiella pneumoniae (Kp1), 4 as Klebsiella quasipneumoniae (Kp2), and 3 as Klebsiella variicola (Kp3). Kp1-Kp3 could not be differentiated using biochemical tests, mass spectrometry analysis, or 16S rRNA gene sequencing, whereas multiplex PCR of β-lactamase genes and tyrB gene sequencing enabled phylogrouping of all three. There were differences in the detection rates of capsular genes (capsule associated gene G and magnesium transport system ATP-binding protein A) and siderophore genes (Klebsiella pneumoniae iron uptake system genes) among Kp1-Kp3; susceptibility to different antibiotics also varied considerably. ConclusionKlebsiella pneumoniae phylogroups isolated from commercially produced laboratory rats and mice are similar to those in human clinical samples and likewise include multiple phylogroups of the species complex. In this study, the identification methods of Kp1-Kp3 were optimized,furthermore selected virulence genes and antimicrobial resistance in Kp1-Kp3 were preliminarily investigated, which could provide precise reference for the treatment of infections caused by different phylogroups occurring in laboratory animal facilities.

Objective:To investigate the clinical characteristics and prognosis of duodenal neuroendocrine neoplasms.Methods:The clinical data of 35 patients with duodenal neuroendocrine neoplasms admitted to Union Hospital, Tongji Medical College, Huazhong University of Science & Technology from Jan 2012 to Dec 2021 were retrospectively analyzed. The differences of clinical characteristics between periampullary and non-periampullary duodenal neuroendocrine neoplasms were analyzed. Kaplan-Meier curve was used for survival analysis, and the clinical factors affecting the prognosis were analyzed.Results:Of the 35 patients, 30 underwent tumor resection, 7 (23%) developed different degree of complications after operation and were improved and discharged after intervention. A total of 5 patients died during the follow-up period. Only 1 of 30 patients who underwent tumor resection died 30 months after operation due to disease progression, and the others had no recurrence or metastasis. Univariate analysis showed that tumor size, tumor grade, and tumor location were associated with the prognosis of patients (all P<0.05), and multivariate analysis showed that patients with tumors located.Away from the ampulla had a significantly better prognosis than those located around the duodenal ampulla ( P<0.01). Conclusions:Patients with duodenal neuroendocrine neoplasms have a good prognosis after complete resection; patients with duodenal neuroendocrine neoplasms located around the ampulla of Vater have a relatively poor prognosis compared with those away from the area of ampulla.

Objective Conduct routine microbiological monitoring of laboratory rats and mice from vendors to provide an important basis for the scientific management of laboratory animal facility and ensure the reliability of relevant experimental data obtained from laboratory animals.Methods Taking the Department of Laboratory Animal Science of Fudan University as an example, between April 2021 and April 2023, rats and mice purchased from 7 vendors were sampled for microbiological quality according to the principle of simple random sampling on the arrival days of animal delivery. Then, surveillance tests were conducted to examine the microbiological contaminations according to the national standards of SPF laboratory animals.Results The total qualified rate was 80.36%, with 52.63% in SD rat, 82.76% in inbred mice, 86.67% in outbred mice and 86.36% in immunodeficient mice in details. The most frequent bacteria isolated were Staphylococcus aureus, Pseudomonas aeruginosa, Klebsilla pneumoniae and Rodentibacter heylii, and their detection rates were 10.76%, 3.16%, 2.53% and 0.63%, respectively. Serological assays demonstrated the highest prevalence for virus was Sendai virus, and the detection rate was 2.53%. In addition to the pathogens those must be excluded from SPF rodents, Entamoeba muris and Enterobacter spp. were also detected in inbred mice, and Klebsiella oxytoca was detected in immunodeficient mice, with the detection rates of 1.15%, 2.30% and 4.55%, respectively.Conclusion There are certain incidences of pathogen infections in laboratory rats and mice from vendors, and an efficient microbiological monitoring of laboratory animals should be implemented in animal facilities, in order to eliminate pathogen infections in laboratory animals, which is required for improving the accuracy of research results and protecting the occupational health of laboratory animal practitioners as well.

OBJECTIVE:To study the effects of 3 extracts of Acanthopanax sessiliflorus fruits on the proliferation and apoptosis of human hepatocarcinoma cells SMMC-7721,and to provide reference for confirming the mechanism of anti-tumor effect. METHODS:MTT assay was adopted to investigate the effects of low-mass concentration,medium-mass concentration and high-mass concentration of ethanol extract(0.92,1.84,3.68 mg/mL),crude polysaccharide extract(0.06,0.12,0.24 mg/mL)and refined polysaccharide extract (0.04, 0.08, 0.16 mg/mL) from A. sessiliflorus fruits on the proliferation and apoptosis of SMMC-7721 cells after treated for 24,36,48 h,respectively. Flow cytometry was used to investigate the effects of 1.84 mg/mL ethanol extract,0.24 mg/mL crude polysaccharide extract and 0.16 mg/mL refined polysaccharide extract on cell cycle and cell apoptosis after treated for 24 h. The above tests were all negative control(only adding cells without drugs). RESULTS:Compared with negative control,3 extracts of A. sessiliflorus fruits could significantly inhibit the proliferation of SMMC-7721 cells (P<0.01),could significantly decrease the percentage of SMMC-7721 cells in G0/G1 and G2/M phase(P<0.01),could significantly increase the percentage of SMMC-7721 cells in S phase (P<0.01) and the apoptosis rate of SMMC-7721 cells (P<0.05);especially the effects of ethanol extract from A. sessiliflorus fruits were the most obvious. CONCLUSIONS:Three extracts of A.sessiliflorus fruits can inhibit the proliferation of human hepatocarcinoma SMMC-7721 cells,block SMMC-7721 cells in S phase and induce the apoptosis of SMMC-7721 cells.

Objective To investigate the clinical efficacy of asymmetric osteotomy via posterior adjacent vertebrae in the treatment of traumatic thoracolumbar kyphotic scoliotic deformity.Methods A retrospective case series study was conducted on the clinical data of 16 patients with traumatic thoracolumbar kyphotic scoliotic deformity admitted to our department from January 2012 to January 2017.There were 10 males and six females,aged (42.5 ±7.6) years (range,20-62 years).According to the location of injured vertebrae,there were two patients with T11,five with T12,six with L1,and three with L2,all of which had scoliosis deformity and obvious low back pain.All patients underwent asymmetric osteotomy via posterior adjacent vertebrae.The operation time and intraoperative bleeding were recorded.The imaging parameters such as kyphosis Cobb angle,scoliosis Cobb angle,distance between C7 plumbline and central sacral vertebral line (C7-CSVL),and distance between C7 plumbline and sagittal vertical axis (SVA) were measured before and after operation.At the same time,the visual analogue scale (VAS),Oswestry dysfunction index (ODI),and SRS-22 scale were used to evaluate the clinical efficacy.The AISA score was used to evaluate the neurological function recovery before and after operation.Results All patients were followed up for (26.3 ± 16.8) months (range,15-65 months).The operation time was (6.0 ± 1.4) hours (range,4.5-9.0 hours),and the intraoperative bleeding was (900.5 ± 360.8)ml (range,800-1600 ml).The kyphosis Cobb angle was improved from (70.3 ± 9.8) °before operation to (12.2 ± 2.9) ° after operation (P ＜ 0.01).The scoliosis Cobb angle was improved from (47.6 ± 11.6) ° before operation to (4.0 ± 0.9) ° after operation (P ＜ 0.01).The C7-CSVL decreased from preoperative (3.1 ±0.8)cm to postoperative (1.2 ±0.4)cm (P ＜0.01),and the SVA decreased from preoperative (5.0 ± 0.9) cm to postoperative (2.9 ± 0.5) cm (P ＜ 0.01).No severe complications such as spinal cord and nerve injury occurred.The VAS decreased from preoperative (6.8 ± 1.0) to (1.9 ± 0.9) points at the last follow up.The ODI decreased from (54.6 ± 4.2) ％ before operation to (8.1 ± 2.5) ％ at the last follow up.The SRS-22 score was increased from (64.6 ±7.5) points before operation to (87.4 ± 3.2) points at the last follow-up.In terms of the ASIA classification,two patients were improved from grade C to grade D after operation,and six patients were improved from grade D to grade E.Conclusion Asymmetric osteotomy via posterior adjacent vertebrae is safe and effective in the treatment of traumatic thoracolumbar scoliosis,with high correction rate of scoliosis and kyphosis at the same time.

Objective To compare and explore the curative effects of elective operation and emergency operation in treating atlantoaxial vertebral segmental spinal canal space-occupying lesions.Methods Thirty-two patients suffering from atlanto-axial vertebral segmental spinal canal space-occupying lesions treated in our hospital from May 2010 to April 2015 were selected and divided into the emergency operation group (group A,n =14) and elective operation group (group B,n =18).The emergency and elective operations were adopted respectively.Then the operation time,intraoperative blood loss,JOA score,ODI index,VAS score,postoperative imaging(MRI) and effect satisfaction degree were compared between the two groups.Results After treatment,the JOA score in the group A was (25.23±4.47) points,which was higher than (22.10±3.56) points in the group B,and the difference was statistically significant (t=3.67,P＜0.05).The ODI index and VAS score of the two groups all were decreased.The ODI index in the group A was (18.56±3.10) points,which in the group B was (21.56±4.37) points,and there was statistically significant difference between the two groups (t=3.76,P＜0.05).The VAS score in the group A was (1.89 ±-0.53)points,which in the group B was (3.16±0.89)points,the difference was statistically significant between the two groups (t=3.76,P＜0.05).Before surgery and at postoperative 1 month,the spinal cord function classification(Frankel grade) of the two groups had no statistically significant difference between the two groups(Z=-0.18,P=0.85＞0.05,Z=-0.52,P=0.60＞0.05).The operation time had no statistical difference between the group A and B[(120.23±9.02)min vs.(126.25±12.12)min,P＞0.05].The intraoperative bleeding volume had had no statistical difference between the group A and B [(211.26±12.25)mL vs.(220.43±17.58)mL,P＞ 0.05].After one month of treatment,the satisfaction degree in the group A was 92.56 ％,which was higher than 72.22％ in the group B,and the difference was statistically significant (Z=-2.13,P＜0.05).Conclusion Emergency operation in treating atlantoaxial segment spinal space occupying lesions can effectively improve the therapeutic effect,and has higher patients satisfaction after treatment.Therefore which is worth promoting and applying.

OBJECTIVE:To compare the pharmacodynamics of active parts in crude Atractylodes lancea and A. lancea fired with bran. METHODS:170 rats were randomly divided into 17 groups,including blank control group,spleen and stomach damp obstruction model group,volatile oil of crude A. lancea and A. lancea fired with bran high-dose and low-dose(0.747,0.083 mg/ml by the concentration of crude medicinal materials,similarly hereinafter)groups,solvent control 2% polysorbate 80 group,dichlo-romethane extract of crude A. lancea and A. lancea fired with bran high-dose and low-dose groups,solvent control 1‰ polysorbate 80 group,n-butyl alcohol extract of crude A. lancea and A. lancea fired with bran high-dose and low-dose groups,solvent control stomach damp obstruction model distilled water control group. Except blank control group,other 16 groups were given Sennae foli-um decoction ig for 14 d to induce spleen and stomach damp obstruction model,and then received relevant medicine or solvent ig once a day for consecutive 7 d. Body weight of rats were determined before and after medication,and the serum levels of amylase, D-xylose,gastrin,vasoactive intestinal peptide and NO were determined after medication. RESULTS:Compared with spleen and stomach damp obstruction model group,the body weight and serum levels of gastrin,amylase and D-xylose increased significantly in rats of active part in A. lancea fired with bran groups(P<0.05),while vasoactive intestinal peptide and NO decreased signifi-cantly(P<0.05). Compared active part in crude A. lancea group,except the body weight,above indicators of active part in A. lan-cea fired with bran groups had greater change,with statistical significance (P<0.05). CONCLUSIONS:There are pharmacody-namic differences in the active parts between crude A. lancea and A. lancea fired with bran,the latter one is stronger.

BACKGROUND: The choice of suitable bone graft substitute is vital for spinal fusion treatment, which can solve some limitations caused by autogenous bone graft and other materials. OBJECTIVE: To investigate properties of different bone graft fusion materials, and to explore their application in dog spinal fusion of lumbar vertebral body. METHODS: Forty-five Chinese rural dogs were enrol ed to prepare lumbar interbody fusion models, and then were randomized into three groups transplanted with autogenous ilium, recombinant human bone morphogenetic protein-2 composite or al ograft ilium, respectively. Afterwards, effects of different materials in the lumbar interbody fusion were analyzed. RESULTS AND CONCLUSION: The fusion rate of the composite group was significantly higher than those of the other groups (P < 0.05). Oswestry dysfunction index of the composite group was significantly lower than those of the other groups after surgery (P < 0.05). Hematoxylin-eosin staining showed that dogs achieved complete bony fusion, continuous bone trabecula gradual y formed, and there was no gap between the transplanted bone and the bone surface in the composite group compared with the other groups at 12 weeks after surgery. These results demonstrate that recombinant human bone morphogenetic protein-2 composites can significantly promote the spinal fusion superior to autologous and al ogeneic bone grafts.

Objective:On the basis of the use of chemical methods to establish mouse model of lupus nephritis and its biological identification , we investigate the reverse effect of pathological lesions of B 7-1 human-mouse chimeric antibody blockade against B7/D28 signaling pathway in mice with lupus nephritis model.Methods:Pristane was injected intraperitoneally to 6-week-old female C57BL/6J mice at dose of 0.5 ml per mouse in one go,and urine protein,ANA and renal pathological changes were detected on a monthly basis.Mice whose urine protein content reached ++and ANA fluorescence intensity reached ++were randomly devided into three groups ,five each.Antibody intervention group was sequentially injected with B 7-1-mouse chimeric antibody by orbital venous , positive control group was injected with immunosuppressant CTX , negative control group was injected with isotype control IgG.Urine protein and ANA were also detected on a monthly basis.Mice were sacrificed three months after intervention was executed.Kidney was used for H&E dying , IC detection and electric microscope observation.Results: After four-month Pristane induction , urine protein content of 80%mice reached +-+++,meanwhile,serum ANA fluorescence intensity reached ++-+++.Glomerulonephritis infiltrating cells were observed Mice with urine protein and ANA , glomerular inflammatory cell infiltration , tubular epithelial cell degeneration visible edema ,vascular congestion significantly ,fibrosis.After antibody intervention ,urine protein content in antibody intervention group gradually reduced from ++-+++to ±-+++,ANA ++-+++to +-++,and were significantly different from that in the negative control group ( P<0.01 ).Analysis of kidney H&E dying showed that antibody glomerular infiltration of inflammatory cells in the intervention group and tubular congestion and other symptoms were improved significantly.Immunofluorescence staining indicated that fluorescence intensity of IC was significantly reduced in the antibody intervention group.Electron dense deposits reduction and glomerular basement membrane uniformity were observed in antibody intervention group by electric microscope when compared with the negative control group.Conclusion:B7-1 antibodies could downregulate immune response through inhibiting B 7-1/CD28 signaling pathway , reducing the production of autoantibodies and reversing pathological damage caused by autoimmune response .

Objective To elucidate whether and how injection site diversity influences on the sodium hyaluronate (HA) distribution on cartilage surface of the knee joint.Methods From September 2014 to December 2014, 32 human cadaveric knees were sorted from small to large based on femoral condyle width (FCW) and divided into group Ⅰ (n=16, odd number, the patella medial approach) and group Ⅱ (n=16, even number, injected through the medial joint line approach).Knees of both groups were interfused with a 5％ (mg: ml) methylene blue tracer.Each specimen was then simulated a 5°-45° alternating walking for 2 h in the Electro Force platform.Afterwards, all the knee cavities were cut open to examine the range of HA distribution.Based on the eight-zoning classification law for the knee cartilage surface, the coverage area of HA in each zone was scored and the characteristics of HA distribution was depicted, as well as HA zonal distribution diversity between groups were statistically analyzed.Results HA of both patella medial and medial joint line subgroups showed analogical distribution in all zones except the lateral tibial plateau at the time of FCW ≤ 7.0 cm.However, HA coverage through the patella medial approach showed significantly higher distribution scores in the patella zone and the femoral trochlear zone, but significant lower distribution scores in the antero-lateral/postero-lateral femoral condyle, the medial posterior femoral condyle zone and the lateral tibial plateau zone compared with medial joint line approach at the time of FCW ＞ 7.0 cm.Conclusion When delivered through different injection approaches, HA showed analogical distribution characteristics in most zones of the knee cavity in population with a small bodily shape (FCW ≤ 7.0cm), but significant diverse distribution characteristics in population with a big bodily shape (FCW ＞7.0 cm).When FCW ＞ 7.0cm, HA injected through the medial-patellar approach tended to be distributed mainly over the patella-femoral articulation and the anterior knee cavity, while HA injected through the joint line approach tended to be distributed mainly over the tibio-femoral articulation and the posterior knee cavity.