ObjectiveTo establish and evaluate different disease-syndrome combination mice models of knee osteoarthritis （KOA） with cold-dampness obstruction syndrome， and to identify the optimal modeling method. MethodsEighty mice were randomly divided into a control group， a treadmill plus climate chamber group， an anterior cruciate ligament transection （ACLT） plus climate chamber group， an ACLT group， a destabilization of the medial meniscus （DMM） plus climate chamber group， a DMM group， an intra-articular injection plus climate chamber group， and an intra-articular injection group， with 10 mice in each group. Mice in the ACLT group underwent surgical transection of the knee anterior cruciate ligament to establish the model， while those in the DMM underwent surgical transection of the medial meniscal tibial ligament of the knee， and those in the intra-articular injection group were modeled by an intra-articular injection of 50 μl of normal saline containing 1mg of monosodium iodoacetate （MIA） into the medial side of the patellar ligament. Mice in the ACLT plus climate chamber， DMM plus climate chamber， and intra-articular injection plus climate chamber groups were placed in an artificial climate chamber （relative humidity 60%， temperature 4 ℃， wind speed 6 m/s） simultaneously with aforementioned interventions， once daily for 2 h each time， for 14 consecutive days. Mice in the control group underwent sham surgery， involving only the incision of the skin and subcutaneous tissue at the knee joint to expose the articular cavity， followed by layer-by-layer suturing. Mice in the treadmill plus climate chamber group were placed on an animal treadmill starting on the 4th day after the sham operation， undergoing high-intensity treadmill exercise at a speed of 20 m/min， an inclination of 5°， for 30min per session， 5 times per week for 28 consecutive days， while being housed in the artificial climate chamber. Cold pain thresholds of the mice in each group were compared using the cold plate test and the acetone low-temperature test before and after modeling. Following modeling， knee joint pathological changes were observed using HE and Safranin O-Fast Green staining， and quantified using Mankin and Osteoarthritis Research Society International （OARSI） scores. Serum levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） were measured by ELISA. Protein and mRNA expression levels of pain-related markers， including calcitonin gene-related peptide （CGRP）， protein gene product 9.5 （PGP9.5）， and cellular proto-oncogene Fos （c-Fos）， and the cold hyperalgesia-related markers including transient receptor potential ankyrin 1 （TRPA1） and transient receptor potential melastatin 8 （TRPM8） in the dorsal root ganglion （DRG） tissues were detected by Western Blot and qPCR. TRPA1 and TRPM8 fluorescent intensity in the DRG was detected by immunofluorescence assay. ResultsCompared to the control group， the remaining groups showed decreased cold pain thresholds in the cold plate tese， increased cold pain thresholds in the acetone low-temperature test， elevated Mankin and OARSI scores， and increased serum TNF-α and IL-1β levels， upregulated protein and mRNA expression levels of CGRP， PGP9.5， c-Fos， TRPA1， and TRPM8 in the DRG， and increased immunofluorescence intensities of TRPA1 and TRPM8 in the DRG tissue （P＜0.05 or P＜0.01）， along with severe pathological damage and cartilage degeneration in the knee joints. Compared to the ACLT， DMM， and intra-articular injection groups， the ACLT plus climate chamber， DMM plus climate chamber， and intra-articular injection plus climate chamber groups exhibited more significant changes in all the aforementioned indicators （P＜0.01）. Furthermore， changes in the aforementioned indicators were more pronounced in the ACLT plus climate chamber group compared to the Treadmill plus climate chamber group， DMM plus climate chamber and intra-articular injection plus climate chamber group （P＜0.01）， exhibiting the most severe degree of pathological damage and cartilage degeneration. ConclusionThe combination of an artificial climate chamber can successfully establish a disease-syndrome combined animal model of KOA with cold-damp obstruction syndrome， with the ACLT combined with the artificial climate chamber being the optimal modeling approach.

ObjectiveTo establish and evaluate different disease-syndrome combination mice models of knee osteoarthritis （KOA） with cold-dampness obstruction syndrome， and to identify the optimal modeling method. MethodsEighty mice were randomly divided into a control group， a treadmill plus climate chamber group， an anterior cruciate ligament transection （ACLT） plus climate chamber group， an ACLT group， a destabilization of the medial meniscus （DMM） plus climate chamber group， a DMM group， an intra-articular injection plus climate chamber group， and an intra-articular injection group， with 10 mice in each group. Mice in the ACLT group underwent surgical transection of the knee anterior cruciate ligament to establish the model， while those in the DMM underwent surgical transection of the medial meniscal tibial ligament of the knee， and those in the intra-articular injection group were modeled by an intra-articular injection of 50 μl of normal saline containing 1mg of monosodium iodoacetate （MIA） into the medial side of the patellar ligament. Mice in the ACLT plus climate chamber， DMM plus climate chamber， and intra-articular injection plus climate chamber groups were placed in an artificial climate chamber （relative humidity 60%， temperature 4 ℃， wind speed 6 m/s） simultaneously with aforementioned interventions， once daily for 2 h each time， for 14 consecutive days. Mice in the control group underwent sham surgery， involving only the incision of the skin and subcutaneous tissue at the knee joint to expose the articular cavity， followed by layer-by-layer suturing. Mice in the treadmill plus climate chamber group were placed on an animal treadmill starting on the 4th day after the sham operation， undergoing high-intensity treadmill exercise at a speed of 20 m/min， an inclination of 5°， for 30min per session， 5 times per week for 28 consecutive days， while being housed in the artificial climate chamber. Cold pain thresholds of the mice in each group were compared using the cold plate test and the acetone low-temperature test before and after modeling. Following modeling， knee joint pathological changes were observed using HE and Safranin O-Fast Green staining， and quantified using Mankin and Osteoarthritis Research Society International （OARSI） scores. Serum levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） were measured by ELISA. Protein and mRNA expression levels of pain-related markers， including calcitonin gene-related peptide （CGRP）， protein gene product 9.5 （PGP9.5）， and cellular proto-oncogene Fos （c-Fos）， and the cold hyperalgesia-related markers including transient receptor potential ankyrin 1 （TRPA1） and transient receptor potential melastatin 8 （TRPM8） in the dorsal root ganglion （DRG） tissues were detected by Western Blot and qPCR. TRPA1 and TRPM8 fluorescent intensity in the DRG was detected by immunofluorescence assay. ResultsCompared to the control group， the remaining groups showed decreased cold pain thresholds in the cold plate tese， increased cold pain thresholds in the acetone low-temperature test， elevated Mankin and OARSI scores， and increased serum TNF-α and IL-1β levels， upregulated protein and mRNA expression levels of CGRP， PGP9.5， c-Fos， TRPA1， and TRPM8 in the DRG， and increased immunofluorescence intensities of TRPA1 and TRPM8 in the DRG tissue （P＜0.05 or P＜0.01）， along with severe pathological damage and cartilage degeneration in the knee joints. Compared to the ACLT， DMM， and intra-articular injection groups， the ACLT plus climate chamber， DMM plus climate chamber， and intra-articular injection plus climate chamber groups exhibited more significant changes in all the aforementioned indicators （P＜0.01）. Furthermore， changes in the aforementioned indicators were more pronounced in the ACLT plus climate chamber group compared to the Treadmill plus climate chamber group， DMM plus climate chamber and intra-articular injection plus climate chamber group （P＜0.01）， exhibiting the most severe degree of pathological damage and cartilage degeneration. ConclusionThe combination of an artificial climate chamber can successfully establish a disease-syndrome combined animal model of KOA with cold-damp obstruction syndrome， with the ACLT combined with the artificial climate chamber being the optimal modeling approach.

Objective To evaluate the impact of different methods of harvesting palatal connective tissue on patients' postoperative pain at the palatal donor site.Methods A total of 30 patients were included.The control group was de-epithelialized free gingival graft technique group,and the experimental group was the single-incision technique group.The keratinized gingival width of the recipient site,number of vertical incisions,number of teeth,donor site mucosa thickness,graft width,graft length,graft thickness,operation time and amount of anesthetic were recorded.The postoperative quality of life rating scale and visual analog scale were completed and statistical analysiswas performedon the results by SPSS 26.0.Results The postoperative quality of life score of the experimental group was significantly lower than that of the control group,and there was no significant difference in the total VAS score of the donor site.In terms of the total postoperative pain level after mucogingival surgery,the number of vertical incisions and graft thickness were positively correlated with the pain.Palate mucosal thickness was negatively correlated with postoperative pain levels.Conclusion In mucogingi-val surgery,the degree of postoperative pain in the donor palatal region of the patient is independent of the choice of de-epithelialized free gingival graft technique or single-incision technique to obtain subepithelial connective tissue.The surgeon should also give compre-hensive considerationsto the anatomical factors of the patient's donor site,surgical technical level and experience,and aesthetic require-ments when selecting the surgical method.

Objective To evaluate the impact of different methods of harvesting palatal connective tissue on patients' postoperative pain at the palatal donor site.Methods A total of 30 patients were included.The control group was de-epithelialized free gingival graft technique group,and the experimental group was the single-incision technique group.The keratinized gingival width of the recipient site,number of vertical incisions,number of teeth,donor site mucosa thickness,graft width,graft length,graft thickness,operation time and amount of anesthetic were recorded.The postoperative quality of life rating scale and visual analog scale were completed and statistical analysiswas performedon the results by SPSS 26.0.Results The postoperative quality of life score of the experimental group was significantly lower than that of the control group,and there was no significant difference in the total VAS score of the donor site.In terms of the total postoperative pain level after mucogingival surgery,the number of vertical incisions and graft thickness were positively correlated with the pain.Palate mucosal thickness was negatively correlated with postoperative pain levels.Conclusion In mucogingi-val surgery,the degree of postoperative pain in the donor palatal region of the patient is independent of the choice of de-epithelialized free gingival graft technique or single-incision technique to obtain subepithelial connective tissue.The surgeon should also give compre-hensive considerationsto the anatomical factors of the patient's donor site,surgical technical level and experience,and aesthetic require-ments when selecting the surgical method.

Objective To observe the volume changes of brain regions and their correlation with cognitive function in temporal lobe epilepsy(TLE)patients.Methods Fifty TLE patients(TLE group)and 34 healthy controls(control group)were prospectively enrolled.Brain 3D-T1WI was collected,brain regions were delineated with automated segmentation technique,and hippocampus was segmented additionally with hippocampal brain template(HBT).The volume of brain regions were compared between groups,and the correlation of brain region volume with Montreal cognitive assessment(MoCA)score were analyzed.Results Compared with control group,in TLE group,bilateral cerebrum white matter,right globus pallidus,left posterior cingulate gyrus,left precentral gyrus,right fusiform gyrus,left cerebellum white matter,corpus callosum,left hippocampus(HBT segmentation and conventional segmentation)and right thalamus volumes decreased significantly(all P＜0.05),while right choroid plexus volume increased significantly(P＜0.05).The volume of left hippocampus(HBT segmentation)and right fusiform gyrus were both positively correlated with MoCA score in TLE group(r=0.335,0.376,P=0.009,0.007).Conclusion Widespread atrophy of brain regions could be seen in TLE patients,which were correlated with cognitive function.

Objective:To analyze the short-term hospital-based transmission characteristics and gene variation of Carbapenem-Resistant Klebsiella pneumoniae (CRKP) by genome-wide technique to provide evidence for transmission control. Methods:The experimental strain was derived from all the CRKP isolated in Affiliated Hospital of North China University of Science and Technology from October 2022 to December 2023. Strain identification and drug susceptibility were tested with VITEK 2-Compact automatic bacterial identification drug susceptibility analyzer or disk method, and the results were interpreted through whole genome sequencing. The ST type, carbapenem resistance gene, virulence factor, and O serotype of the collected strains were analyzed.Results:Among the 115 strains of CRKP, 94 strains were isolated from the intensive care unit (ICU), accounting for 81.7%, and 21 strains were isolated from the non-intensive care unit (NICU), accounting for 18.3%. The 115 strains of CRKP can be divided into 11 ST types, of which ST11 type was the most (54.8%, 63/115), followed by ST15 type (22.6%, 26/115) and ST5492 type (15.7%, 18/115). Type ST5492 was a new clonal group in the region. The 115 strains of CRKP could be divided into 7 O serotypes, most of which were O2a type(32.2%,37/115), followed by O5 type(30.4%,35/115) and O1 type(27.8%,32/115). The resistance genes of carbapenem antibiotics showed that there were 107 strains carrying the blaKPC-2 gene, one strain with the blaNDM-1 gene, and one strain with both the blaKPC-2 and blaNDM-13 genes. Virulence genes were detected in 55 CRKP strains (47.8%, 55/115), among which six strains detected peg-344, iucA, iroB, rmpA, and rmpA2 virulence genes (5.2%, 6/115). Four virulence genes ( peg-344, iucA, rmpA, and rmpA2) were detected in 34 strains (29.6%, 34/115). Three virulence genes ( iucA, iroB and rmpA) were detected in two strains (1.7%, 2/115). Three virulence genes ( peg-344, iucA and rmpA) were detected in one strain (0.8%, 1/115). IucA and rmpA virulence genes were detected in 12 strains (10.4%, 12/115). KPC-2_ST11_O2a, KPC-2_ST15_O1 and KPC-2_ST5492_O5 were dominant clones, and their distribution was mainly in the intensive care unit. The whole genome sequence analysis showed that there were three dominant clones, among which ST11 clones were subdivided into three dominant O serotypes, all of which were mainly in the intensive care unit. Conclusion:The popular strain in the hospital of CRKP is a KPC-2_ST11 clone group carrying iucA, rmpA/rmpA2, with cross-department transmission and mutation. ST5492 is a newly-launched clone type. The intensive care unit of hvKP carrying five virulence genes, including peg-344, should be alert to the epidemic risk of CR-hvKP outbreak.

Objective To observe the volume changes of brain regions and their correlation with cognitive function in temporal lobe epilepsy(TLE)patients.Methods Fifty TLE patients(TLE group)and 34 healthy controls(control group)were prospectively enrolled.Brain 3D-T1WI was collected,brain regions were delineated with automated segmentation technique,and hippocampus was segmented additionally with hippocampal brain template(HBT).The volume of brain regions were compared between groups,and the correlation of brain region volume with Montreal cognitive assessment(MoCA)score were analyzed.Results Compared with control group,in TLE group,bilateral cerebrum white matter,right globus pallidus,left posterior cingulate gyrus,left precentral gyrus,right fusiform gyrus,left cerebellum white matter,corpus callosum,left hippocampus(HBT segmentation and conventional segmentation)and right thalamus volumes decreased significantly(all P＜0.05),while right choroid plexus volume increased significantly(P＜0.05).The volume of left hippocampus(HBT segmentation)and right fusiform gyrus were both positively correlated with MoCA score in TLE group(r=0.335,0.376,P=0.009,0.007).Conclusion Widespread atrophy of brain regions could be seen in TLE patients,which were correlated with cognitive function.

Objective:To analyze the short-term hospital-based transmission characteristics and gene variation of Carbapenem-Resistant Klebsiella pneumoniae (CRKP) by genome-wide technique to provide evidence for transmission control. Methods:The experimental strain was derived from all the CRKP isolated in Affiliated Hospital of North China University of Science and Technology from October 2022 to December 2023. Strain identification and drug susceptibility were tested with VITEK 2-Compact automatic bacterial identification drug susceptibility analyzer or disk method, and the results were interpreted through whole genome sequencing. The ST type, carbapenem resistance gene, virulence factor, and O serotype of the collected strains were analyzed.Results:Among the 115 strains of CRKP, 94 strains were isolated from the intensive care unit (ICU), accounting for 81.7%, and 21 strains were isolated from the non-intensive care unit (NICU), accounting for 18.3%. The 115 strains of CRKP can be divided into 11 ST types, of which ST11 type was the most (54.8%, 63/115), followed by ST15 type (22.6%, 26/115) and ST5492 type (15.7%, 18/115). Type ST5492 was a new clonal group in the region. The 115 strains of CRKP could be divided into 7 O serotypes, most of which were O2a type(32.2%,37/115), followed by O5 type(30.4%,35/115) and O1 type(27.8%,32/115). The resistance genes of carbapenem antibiotics showed that there were 107 strains carrying the blaKPC-2 gene, one strain with the blaNDM-1 gene, and one strain with both the blaKPC-2 and blaNDM-13 genes. Virulence genes were detected in 55 CRKP strains (47.8%, 55/115), among which six strains detected peg-344, iucA, iroB, rmpA, and rmpA2 virulence genes (5.2%, 6/115). Four virulence genes ( peg-344, iucA, rmpA, and rmpA2) were detected in 34 strains (29.6%, 34/115). Three virulence genes ( iucA, iroB and rmpA) were detected in two strains (1.7%, 2/115). Three virulence genes ( peg-344, iucA and rmpA) were detected in one strain (0.8%, 1/115). IucA and rmpA virulence genes were detected in 12 strains (10.4%, 12/115). KPC-2_ST11_O2a, KPC-2_ST15_O1 and KPC-2_ST5492_O5 were dominant clones, and their distribution was mainly in the intensive care unit. The whole genome sequence analysis showed that there were three dominant clones, among which ST11 clones were subdivided into three dominant O serotypes, all of which were mainly in the intensive care unit. Conclusion:The popular strain in the hospital of CRKP is a KPC-2_ST11 clone group carrying iucA, rmpA/rmpA2, with cross-department transmission and mutation. ST5492 is a newly-launched clone type. The intensive care unit of hvKP carrying five virulence genes, including peg-344, should be alert to the epidemic risk of CR-hvKP outbreak.

Severe acute pancreatitis(SAP)is a critical condition in general surgery settings,characterized by high mortality and poor prognosis.On February 28,2024,the Department of Hepatobiliary Surgery at the First Affiliated Hospital of Harbin Medical University admitted a 36-year-old male patient.The patient presented with"upper abdominal pain accompanied by fever for three months and jaundice of the skin and sclera for one week."Physical examination revealed 11 puncture tubes,and a palpable mass measuring 3 cm × 5 cm in the upper abdomen.Enhanced CT and magnetic resonance cholangiopancreatography indicated acute pancreatitis.The patient was diagnosed with"SAP,infectious pancreatic necrosis,and biliary stenosis."He had severe abdominal infection and complex postoperative complications,making treatment challenging.Consequently,a multidisciplinary team(MDT)consultation was initiated.After three rounds of MDT consulfation and freating,the patient ultimately recovered successfully and was discharged.This article reviews the MDT treatment process for this patient and summarizes the characteristics of this condition based on relevant literature to provide insights and experience for clinical practice.