To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

The receptors of Newcastle disease virus(NDV)are sialic acid receptors that mainly in-clude neu5ac-α-2,3gal-β-1,4Glc(SAα2,3Gal)and neu5ac-2-s-α-2,6Gal10Me(SAα2,6Gal).The distribution and abundance of the two receptors in host cells have important effects on virus sus-ceptibility and intracellular proliferation.In order to further explore the effects of sialic acid recep-tors on susceptibility and proliferation characteristics of NDV different strains,the expression lev-els of SAα2,3Gal and SAα2,6Gal receptors on BHK-21 cell membrane were adjusted by overex-pression and RNAi assays,and the TCID50 values were determined after different BHK-21 cells were inoculated with NDV strains Ⅰ and LaSota.The results suggested that NDV strain LaSota preferentially binds to SAα2,6Gal and strain Ⅰ selectively binds to SAα2,3Gal receptor.Further-more,the viral titers of NDV strains LaSota and Ⅰ in cell culture were positively correlated with the expression levels of SAα2,6Gal and SAα2,3Gal receptors on host cell membrane respectively.In conclusion,our studies provide an understanding of the relationship between infectivity of NDV different strains and receptor types of host cell,and provide a method to increase viral titer of NDV for cell-based vaccine production.

Objective To explore the mechanism of GS-9620 in improving imiquimod(IMQ)-induced psoriasis-like inflammation by regulating the Th1/Th17-related immune response,and to investigate its regulatory effect on the gut microbiota in mice.Methods An IMQ-induced psoriasis-like inflammation model was established in BALB/c mice.The severity of the skin lesions was evaluated by psoriasis area and severity index(PASI)score.The proportions of CD4+interleukin(IL)-17+and CD4+interferon(IFN)-γ+cells in spleen tissue were detected by flow cytometry.Levels of the inflammatory factors tumor necrosis factor(TNF)-α,IL-1β,and IL-6 in skin tissues were determined by enzyme-linked immunosorbent assay,and pathological analysis was performed by hematoxylin/eosin staining.The effects of GS-9620 on the structure of the gut microbiota in control,IMQ model,and GS-9620-treated mice were detected by 16S rRNA sequencing.Results GS-9620 significantly reduced the PASI score in IMQ-induced mice and effectively reduced the proportions of CD4+IL-17+and CD4+IFN-γ+cells in the spleen.GS-9620 also significantly down-regulated the expression levels of TNF-α,IL-1β,and IL-6 in skin tissues.16S rRNA sequencing showed that GS-9620 significantly regulated the abundance of gut microbiota related to inflammation,including the relative abundances of bacteria such as Lachnospiraceae_NK4A136_group,Lachnospiraceae_UCG-008,Alloprevotella,Desulfovibrio,Prevotellaceae_UCG-001,and Alistipes.Conclusions GS-9620 effectively alleviates IMQ-induced psoriasis-like skin inflammation in mice by regulating the expression of Th1/Th17-related inflammatory factors.It may also improve IMQ-induced clinical symptoms by regulating the structure of the gut microbiota,thus providing a new theoretical basis for the treatment of psoriasis.The result of this study provide important experimental evidence to support further investigations into the application of GS-9620 for the treatment of psoriasis.

Objective To explore the mechanism of GS-9620 in improving imiquimod(IMQ)-induced psoriasis-like inflammation by regulating the Th1/Th17-related immune response,and to investigate its regulatory effect on the gut microbiota in mice.Methods An IMQ-induced psoriasis-like inflammation model was established in BALB/c mice.The severity of the skin lesions was evaluated by psoriasis area and severity index(PASI)score.The proportions of CD4+interleukin(IL)-17+and CD4+interferon(IFN)-γ+cells in spleen tissue were detected by flow cytometry.Levels of the inflammatory factors tumor necrosis factor(TNF)-α,IL-1β,and IL-6 in skin tissues were determined by enzyme-linked immunosorbent assay,and pathological analysis was performed by hematoxylin/eosin staining.The effects of GS-9620 on the structure of the gut microbiota in control,IMQ model,and GS-9620-treated mice were detected by 16S rRNA sequencing.Results GS-9620 significantly reduced the PASI score in IMQ-induced mice and effectively reduced the proportions of CD4+IL-17+and CD4+IFN-γ+cells in the spleen.GS-9620 also significantly down-regulated the expression levels of TNF-α,IL-1β,and IL-6 in skin tissues.16S rRNA sequencing showed that GS-9620 significantly regulated the abundance of gut microbiota related to inflammation,including the relative abundances of bacteria such as Lachnospiraceae_NK4A136_group,Lachnospiraceae_UCG-008,Alloprevotella,Desulfovibrio,Prevotellaceae_UCG-001,and Alistipes.Conclusions GS-9620 effectively alleviates IMQ-induced psoriasis-like skin inflammation in mice by regulating the expression of Th1/Th17-related inflammatory factors.It may also improve IMQ-induced clinical symptoms by regulating the structure of the gut microbiota,thus providing a new theoretical basis for the treatment of psoriasis.The result of this study provide important experimental evidence to support further investigations into the application of GS-9620 for the treatment of psoriasis.

To verify whether chaperones can promote the soluble expression of PlpE in Escherichia coli and whether the expressed protein is active,prokaryotic expression and Western blot detection were performed.The results showed that:The PlpE prokaryotic expression vector pET-32a(+)-plpE was expressed as inclusion body,and the expression form was not changed by changing the concentration of inducer,induction time and temperature.The companion proteins pG-KJE8,pGro7,pKJE7 and pG-Tf2 were co-expressed with pET-32a(+)-plpE in Eschierichia coli expres-sion system,respectively.When the final concentration of IPTG of 0.5 mmol/L,L-arabinose of 0.5 g/L or tetracycline of 5.0 μg/L were added as inducers and induced at 37 ℃ for 8 h,the results showed that the molecular companion pGro7 could change the expression of rp-PlpE from inclu-sion body to soluble expression.pG-KJE8,pKJE7 and pG-Tf2 had no effect on the expression of rp-PlpE.The soluble rp-PlpE can react specifically with the positive serum of Mannheimia haemolyti-ca.Therefore,the study showed that the co-expression of the chaperone protein pGro7 can make the rp-PlpE protein express in a soluble form,and the purified protein exhibits reactogenicity.These findings lay the foundation for the establishment of a subunit vaccine and serological diagno-sis methods for Mannheimia haemolytica.

The receptors of Newcastle disease virus(NDV)are sialic acid receptors that mainly in-clude neu5ac-α-2,3gal-β-1,4Glc(SAα2,3Gal)and neu5ac-2-s-α-2,6Gal10Me(SAα2,6Gal).The distribution and abundance of the two receptors in host cells have important effects on virus sus-ceptibility and intracellular proliferation.In order to further explore the effects of sialic acid recep-tors on susceptibility and proliferation characteristics of NDV different strains,the expression lev-els of SAα2,3Gal and SAα2,6Gal receptors on BHK-21 cell membrane were adjusted by overex-pression and RNAi assays,and the TCID50 values were determined after different BHK-21 cells were inoculated with NDV strains Ⅰ and LaSota.The results suggested that NDV strain LaSota preferentially binds to SAα2,6Gal and strain Ⅰ selectively binds to SAα2,3Gal receptor.Further-more,the viral titers of NDV strains LaSota and Ⅰ in cell culture were positively correlated with the expression levels of SAα2,6Gal and SAα2,3Gal receptors on host cell membrane respectively.In conclusion,our studies provide an understanding of the relationship between infectivity of NDV different strains and receptor types of host cell,and provide a method to increase viral titer of NDV for cell-based vaccine production.

Objective To analyze the characteristics of hepatitis B infection time in Baqiao Distract of Xian City, predict the incidence trend of hepatitis B in the next years with ARIMA model, and provide theoretical guidance for the adjustment of hepatitis B prevention and control strategies. Methods A total of 7173 cases of hepatitis B in our hospital from 2018 to 2023 were collected as subjects,the annual percentage of change (APC) was used to analyze the characteristics of time of hepatitis B infection in the next years, and the ARIMA optimal model was used to predict the incidence rate in 2024. Results The gross incidence of hepatitis B in Baqiao Distract of Xian City from 2018 to 2023 was 51.30 per 100 000, and the standardized incidence rate was 42.11 per 100 000. The six-year incidence rate showed an upward trend (APC=8.71%,95% CI：3.29% -9.61% , P<0.05). Chi square results showed that the standardized incidence rate of hepatitis B in men is 1.51 times than women, and the incidence rate of hepatitis B in the 30-45 age group was the highest. The prediction results showed that the number of hepatitis B cases in 2024 was 1590, which was 1.91% lower than that in 2023 (1621 cases). Conclusion The infection of hepatitis B is on the rise in Baqiao Distract of Xian City, ARIMA can be used to predict this kind of infectious disease and provide scientific guidance for the prevention of this disease.

Objective Explore differences in the cognitive abilities of socially different Labradors.Methods The dog mentality assessment(DMA)test created by the Swedish Working Dog Association was modified to employ 12 behavioral variables from five subtests of the DMA test,social contact,play Ⅰ,distance-play,ghosts and play Ⅱ,to assess sociability of the dogs.In accordance with the scoring criteria,49 labradors provided by the China Guide Dog Training Centre in Dalian were scored on the social behavioral variables and classified into high(n=15)and low(n=34)sociability groups by cluster analysis.A new system to test canine cognitive ability was developed using the dog cognitive development battery,which tests various domains of cognitive ability such as social cue use,unsolvable task,inhibitory control,cognitive flexibility,working memory,and multistep problem solving task.The dogs'behavioral performance and duration of the test were also recorded.Statistical analysis was performed to determine assess differences in the cognitive abilities of socially diverse dogs.Results Dogs in the high and low social subgroups differed significantly in behavioral variables of the unsolvable task,inhibitory control test,and multistep problem solving task.In the unsolvable task,dogs in the high social group looked at people for significantly longer than dogs in the low social grouping(P=0.008)and looked at people with significantly less latency time than dogs in the low social group(P=0.0001).In the inhibitory control,dogs in the high social group chose significantly more correctly than dogs in the low social group(P=0.034)and chose for significantly less time than dogs in the low social group(P=0.039).In the multistep problem solving task,for dogs in the high social group.successfully completed number of stakes was significantly higher than for dogs in the low social group(P=0.044).The percentage of operation pale time was significantly lower than for dogs in the low social group(P=0.05).The average latency time to solve the bone task was significantly higher than for dogs in the low social group(P=0.037).Moreover,the percentage of operation bone time was significantly lower than for dogs in the low social group(P=0.038).In tests involving a manipulable apparatus,dogs in the high social group spent more time looking at people than dogs in the low social group and less time manipulating the apparatus than dogs in the low subgroup,but no statistically significant differences were observed(P＞0.05).Conclusions Highly sociable labradors have a greater cognitive ability,they are more able to suppress impulses during tests,more able to complete the multistep problem solving task,and more inclined to change strategies to seek new cues from people rather than obsessing over manipulating the apparatus when they are unable to solve a problem.

Objective To construct and identify a patient-derived organoid model,and to investigate the sensitivity of chemotherapy drugs using this model.Methods Pancreatic cancer cells were obtained from the surgical specimens of two female patients with a confirmed diagnosis of pancreatic cancer after tumor tissue digestion,and then the cells were inoculated into a culture dish using matrigel for three-dimensional culture.Paraffin sections were prepared for HE staining and immunohistochemical staining and were compared with the parent tumor tissue to determine whether the histopathological features of the tumor in vivo were preserved.The pancreatic cancer organoids were treated with seven chemotherapy drugs at different concentrations;Cell Titer-Glo?3D reagent was used to measure cell viability,and the results of drug sensitivity were analyzed.Results Two patient-derived pancreatic cancer organoids were successfully constructed,and HE staining and immunohistochemical staining showed that the pancreatic cancer organoids had consistent histopathological features with the tumors of the corresponding patient.Both pancreatic cancer organoids were more sensitive to gemcitabine monotherapy and the combination of oxaliplatin+SN38+fluorouracil,and patient 1 was more sensitive than patient 2.There were individual differences in the response to drugs between the organoids from different patients.Conclusion The pancreatic cancer organoid model successfully constructed in this study can reflect the histological classification of parent pancreatic tumors and can be used for in vitro chemotherapy drug sensitivity test,which is expected to provide a reference for clinical medication.

Objective To explore the establishment of a prognostic model based on machine learning algorithm to predict primary graft dysfunction(PGD)in patients with idiopathic pulmonary fibrosis(IPF)after lung transplantation.Methods Clinical data of 226 IPF patients who underwent lung transplantation were retrospectively analyzed.All patients were randomly divided into the training and test sets at a ratio of 7∶3.Using regularized logistic regression,random forest,support vector machine and artificial neural network,the prognostic model was established through variable screening,model establishment and model optimization.The performance of this prognostic model was assessed by the area under the receiver operating characteristic curve(AUC),positive predictive value,negative predictive value and accuracy.Results Sixteen key features were selected for model establishment.The AUC of the four prognostic models all exceeded 0.7.DeLong and McNemar tests found no significant difference in the performance among different models(both P＞0.05).Conclusions Based on four machine learning algorithms,the prognostic model for grade 3 PGD after lung transplantation is preliminarily established.The overall prediction performance of each model is similar,which may predict the risk of grade 3 PGD in IPF patients after lung transplantation.