To investigate the genomic features and perform cluster analysis of Carbapenem-resistant Klebsiella pneumoniae (CRKP) to provide an experimental basis for guiding the prevention and treatment of CRKP infections.A retrospective case-cohort study was conducted on 19 non-redundant CRKP strains isolated from the Tenth Affiliated Hospital of Southern Medical University between January and June 2023. Whole genome sequencing (WGS) and multilocus sequence typing (MLST) were performed to compare genomic features and analyze the resistance genes and homology of the strains.The results showed that the 19 CRKP strains were isolated from 8 different clinical departments, mainly from respiratory specimens. The whole genome sequencing revealed that the genomic lengths of CRKP ranged from 4.90 to 5.85 Mbp, with contigs N50 values>20 kb for each genome. The median overall GC content was 57.0% (50.4%-57.1%). Comparative genomic analysis identified three regions with high genomic variability. WGS detected 32 resistance genes across 11 categories. All 19 strains carried carbapenem resistance genes ( blaKPC-2 and blaOXA-48), blaTEM-1B extended-spectrum β-lactamase resistance genes, qnrS1 quinolone resistance gene, and fosA fosfomycin resistance gene, with each strain carrying only one carbapenemase gene. The detection rate of blaKPC-2 was 94.7% (18/19). MLST identified three sequence types: ST11, ST437 and ST147, with ST11 being predominant (89.5%, 17/19). Clustering analysis based on acquired resistance genes revealed three clonal transmission patterns among strains 72 and 90, and strains 88, 84, 66 and 79.In conclusion, CRKP strains carry multiple resistance genes, and clustering analysis indicating that nosocomial clonal transmission is closely related to acquired resistance genes. The ST11- blaKPC-2 type strain is the predominant clone. Strengthened surveillance and effective control strategies are necessary to reduce nosocomial transmission of CRKP.

Objective To establish preoperative and postoperative femoral-pelvic-lumbar spine models of patients with developmental dysplasia of the hip(DDH)and healthy volunteers and to study the biomechanical effects of curved periacetabular osteotomy on the lumbar spine.Methods Preoperative and postoperative femoral-pelvic-lumbar spine DICOM data from four patients with DDH and one healthy volunteer were acquired using CT scanning technology,and a three-dimensional finite element model was constructed.The offset method was used to divide the cortical and cancellous bones in Geomagic and the lumbar cartilage,sacroiliac joint,pubic symphysis,and other cartilages were added to SolidWorks.The model was analyzed using ANSYS for finite element analysis,and the gait was the mid-stage of single-leg support during slow walking.The biomechanical changes in the lumbar spine of patients with DDH before and after surgery were analyzed and compared,and the biomechanical data of the lumbar spine of patients after surgery were compared with those of healthy volunteers.Results The femoral-pelvic-lumbar spine models of four patients and a healthy volunteer were established.The results obtained by the established models under each working condition were within the range of the referenced literature,and the validity of the models was proved.The postoperative stresses on the lumbar spine,femoral neck,annulus fibrosus,and nucleus pulposus were much smaller than those of the patients in the preoperative state,and the postoperative stresses on the lumbar spine,femoral neck,annulus fibrosus,and nucleus pulposus of the patients were similar to those of healthy volunteers.Conclusions Curved periacetabular osteotomy significantly reduced the stresses on the lumbar spine and intervertebral discs.Additionally,the stresses on the annulus fibrosus were more uniform after surgery,which indicated that curved periacetabular osteotomy will adjust patients to a healthy state.This study provides a biomechanical basis for the clinical treatment of DDH and helps optimize surgical plans.

To investigate the genomic features and perform cluster analysis of Carbapenem-resistant Klebsiella pneumoniae (CRKP) to provide an experimental basis for guiding the prevention and treatment of CRKP infections.A retrospective case-cohort study was conducted on 19 non-redundant CRKP strains isolated from the Tenth Affiliated Hospital of Southern Medical University between January and June 2023. Whole genome sequencing (WGS) and multilocus sequence typing (MLST) were performed to compare genomic features and analyze the resistance genes and homology of the strains.The results showed that the 19 CRKP strains were isolated from 8 different clinical departments, mainly from respiratory specimens. The whole genome sequencing revealed that the genomic lengths of CRKP ranged from 4.90 to 5.85 Mbp, with contigs N50 values>20 kb for each genome. The median overall GC content was 57.0% (50.4%-57.1%). Comparative genomic analysis identified three regions with high genomic variability. WGS detected 32 resistance genes across 11 categories. All 19 strains carried carbapenem resistance genes ( blaKPC-2 and blaOXA-48), blaTEM-1B extended-spectrum β-lactamase resistance genes, qnrS1 quinolone resistance gene, and fosA fosfomycin resistance gene, with each strain carrying only one carbapenemase gene. The detection rate of blaKPC-2 was 94.7% (18/19). MLST identified three sequence types: ST11, ST437 and ST147, with ST11 being predominant (89.5%, 17/19). Clustering analysis based on acquired resistance genes revealed three clonal transmission patterns among strains 72 and 90, and strains 88, 84, 66 and 79.In conclusion, CRKP strains carry multiple resistance genes, and clustering analysis indicating that nosocomial clonal transmission is closely related to acquired resistance genes. The ST11- blaKPC-2 type strain is the predominant clone. Strengthened surveillance and effective control strategies are necessary to reduce nosocomial transmission of CRKP.

Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

OBJECTIVE: To establish a finite element model of the knee joint based on coronal plane alignment of the knee (CPAK) typing method, and analyze the biomechanical characteristics of different types of knee joints. METHODS: The finite element models of the knee joint were established based on CT scan data of 6 healthy volunteers. There were 5 males and 1 female with an average age of 24.2 years (range, 23-25 years). There were 3 left knees and 3 right knees. According to the CPAK typing method, the knees were rated as types Ⅰ to Ⅵ. Under the same material properties, boundary conditions, and axial loading, biomechanical simulations were performed on the finite element model of the knee joint. Based on the Von Mises stress nephogram and displacement nephogram, the peak stresses of the meniscus, femoral cartilage, and tibial cartilage, and the displacement of the meniscus were compared among different types of knee joints. RESULTS: The constructed finite element model of the knee joint was verified to be effective, and the stress and displacement results were consistent with previous literature. Under the axial load of 1 000 N, the stress nephogram showed that the stress distribution of the medial and lateral meniscus and tibial cartilage of CPAK type Ⅲ knee joint was the most uneven. The peak stresses of the lateral meniscus and tibial cartilage were 9.969 6 MPa and 2.602 7 MPa, which were 173% and 165% of the medial side, respectively. The difference of peak stress between the medial and lateral femoral cartilage was the largest in type Ⅳ knee joint, and the medial was 221% of the lateral. The displacement nephogram showed that the displacement of the medial meniscus was greater than that of the lateral meniscus except for types Ⅲ and Ⅵ knee joints. The difference between medial and lateral meniscus displacement of type Ⅲ knee joint was the largest, the lateral was 170% of the medial. CONCLUSION In the same type of joint line obliquity (JLO), the medial and lateral stress distribution of the knee was more uniform in varus and neutral positions than in valgus position. At the same time, the distal vertex of JLO subgroup can help to reduce the uneven medial and lateral stress distribution of varus knee, but increase the uneven distribution of valgus knee.
Humans ; Finite Element Analysis ; Knee Joint/diagnostic imaging* ; Female ; Biomechanical Phenomena ; Adult ; Male ; Young Adult ; Stress, Mechanical ; Weight-Bearing/physiology* ; Computer Simulation ; Tomography, X-Ray Computed/methods* ; Cartilage, Articular/physiology* ; Range of Motion, Articular ; Menisci, Tibial/anatomy & histology* ; Tibia/anatomy & histology* ; Meniscus/diagnostic imaging* ; Femur/diagnostic imaging* ; Models, Biological

Pleomorphic adenoma (PA) is the most common benign tumour in the salivary gland and has high morphological complexity. However, the origin and intratumoral heterogeneity of PA are largely unknown. Here, we constructed a comprehensive atlas of PA at single-cell resolution and showed that PA exhibited five tumour subpopulations, three recapitulating the epithelial states of the normal parotid gland, and two PA-specific epithelial cell (PASE) populations unique to tumours. Then, six subgroups of PASE cells were identified, which varied in epithelium, bone, immune, metabolism, stemness and cell cycle signatures. Moreover, we revealed that CD36⁺ myoepithelial cells were the tumour-initiating cells (TICs) in PA, and were dominated by the PI3K-AKT pathway. Targeting the PI3K-AKT pathway significantly inhibited CD36⁺ myoepithelial cell-derived tumour spheres and the growth of PA organoids. Our results provide new insights into the diversity and origin of PA, offering an important clinical implication for targeting the PI3K-AKT signalling pathway in PA treatment.
Humans ; Adenoma, Pleomorphic/genetics* ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Transcriptome ; Myoepithelioma

OBJECTIVE To establish the fingerprint of Shenfukang Ⅱ capsule(SC-Ⅱ), and to screen out its indicative compounds for quality control combined with chemometrics methods and network pharmacology. METHODS The HPLC fingerprint of 10 batches of SC-Ⅱ was established, and similarity evaluation was analyzed by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 A Edition) to determine common peaks; common peaks were identified through standard comparison. Chemometrics methods was used to evaluate quality of 10 batches of SC-Ⅱ, and network pharmacology was used to screen out core targets and pathways of SC-Ⅱ. Combined with the above results, indicative compounds for quality control of SC-Ⅱ were screened out. RESULTS A total of 37 common peaks were obtained in the HPLC fingerprint, the similarity of samples was greater than 0.97. Twenty compounds were identified as morroniside, loganin, paeoniflorin and et al. The samples were divided into two categories by chemical pattern recognition, salvianolic acid B, morroniside, salvianolic acid A and paeoniflorin were differential marker compounds for SC-Ⅱ. Network pharmacology predicted that active compounds such as salvianolic acid B, paeoniflorin and morroniside might exert pharmacological effects through 45 core targets and 15 main pathways. The research preliminary preliminarily predicted that morroniside, paeoniflorin and salvianolic acid B were quality control index components for SC-II. CONCLUSION The established HPLC fingerprint method is simple and good repeatability. The quality control indicative compounds of SC-Ⅱ can provide a basis for its quality control.

Facial paralysis causes both physical pain and psychological distress to patients. It is difficult for a patient with facial paralysis to engage with a normal social life and at work. Progresses have been made in recent years in the treatment of facial paralysis. More attentions have been caught by masseteric to facial nerve transposition, which has advantages of adjacency in location, abundancy in nerve supply and reliability in the outcome and now has deemed an important option of facial reanimation. It has not been long since the application of the technique of masseteric to facial nerve transposition in China, therefore it still lacks a universal guidance on practice. In order to achieve the aim of better quality control and popularisation of the technique, hereby a consensus with suggestions on facial reanimation with masseteric to facial nerve transposition is proposed as the reference for surgeons specialised in facial reanimation. This consensus is proposed, discussed and drafted by experts from plastic and reconstructive surgery, oral and maxillofacial surgery, head and neck surgery and neurosurgery.

Objective:Based on the principle that the aggregation-induced emission (AIE) fluorescent probe 6PD-DPAN could bind and aggregate with bacteria, and the fluorescence intensity could reflect the quantity of bacteria, a new method for rapid, convenient, and accurate bacterial drug sensitivity testing was established, which provided a basis for rapid and accurate clinical drug use.Methods:This was a methodological evaluation study. A total of 107 clinical isolates were collected from Houjie Hospital of Dongguan City from January to December 2022, among which 46 isolates were used for the establishment of the new method, and 61 isolates were used for methodological validation. The minimum inhibitory concentration (MIC) determined by broth microdilution method was used as the gold standard, and three antibacterial drugs, gentamicin, levofloxacin, and cefotaxime, were used as experimental drugs. The AIE plate was incubated for 4 hours, and the fluorescence intensity was measured every half an hour to draw a fluorescence change curve. The MIC results were compared with the CLSI breakpoints to determine the bacteria as sensitive, intermediate, or resistant. To simplify the detection process, the ratio of fluorescence intensity at 4 hours(R) was calculated, and the ROC curve was used to analyze the efficacy of R in determining bacterial growth and establish its cutoff value. The new method was used to determine the MIC of 61 clinical isolates, with broth microdilution method as the gold standard. The basic consistency, categorical consistency, very major errors, and major errors of the new method were analyzed, and the consistency between the two methods was determined by the Kappa test.Results:ROC curve analysis of the R after 4 hours of culture: The cut-off value was 3.0, with both sensitivity and specificity for determining bacterial growth being 100%. The median (interquartile) R for bacterial growth inhibition was 11.1 (8.6, 14.4); the median R-value for bacterial growth was 1.1 (1.0, 1.2). Compared to the gold standard, the newly established method showed 100% (61/61) essential agreement in detecting MICs of 61 clinical isolates, with a categorical agreement of 96.7% (59/61). There were no very major or major errors, and the Kappa value was 0.94, indicating good consistency between the newly established method and the microbroth dilution method.Conclusions:This study successfully established a new method for bacterial drug sensitivity testing based on AIE technology, which could obtain satisfactory results within 5 hours, providing a basis for early precision drug treatment in clinical practice.