ObjectiveTo investigate the effects of Schisandrae Chinensis Fructus lignans on schizophrenia induced by dizocilpine maleate （MK-801） in mice and to clarify its mechanism. MethodsMale mice of 4-6 weeks old were randomized into blank， model， positive drug， and low-， medium-， and high-dose （40， 80， 160 mg·kg^-1， respectively） Schisandrae Chinensis Fructus lignans groups. The blank group was administrated with distilled water， and the other groups were injected with 0.5 mg·kg^-1 MK-801 to induce schizophrenia symptoms. Meanwhile， risperidone was injected at 0.2 mg·kg^-1 in the positive drug group， and mice in the intervention groups were injected with corresponding drugs for 14 consecutive days. The behavioral changes of mice were observed by autonomous activity test， open field test， forced swimming test， and water maze test. The levels of dopamine （DA） and 5-hydroxytryptamine （5-HT） in the brain and tumor necrosis factor-α （TNF-α） and nuclear factor-κB （NF-κB） in peripheral blood were quantified by enzyme-linked immunosorbent assay （ELISA）. The changes in the prefrontal lobe of mice were observed by hematoxylin-eosin staining， and the changes of the hippocampal tissue were observed by Nissl staining. The protein levels of silencing information regulatory factor 1 （SIRT1） and forkhead box protein O3a （FoxO3a） in the hippocampus of mice were determined by Western blot. ResultsCompared with the model group， low， medium， and high doses of Schisandrae Chinensis Fructus lignans reduced the total number of autonomous activities， total distance in the open field test， immobile time in the forced swimming test， and levels of TNF-α and NF-κB in peripheral blood （P<0.05）， while increasing the number of platform crossings in the water maze test and DA and 5-HT levels in the brain tissue （P<0.05）. Compared with the model group， risperidone and low， medium， and high doses of Schisandrae Chinensis Fructus lignans improve the neural cell morphology in the CA1 region， with full cells in neatly dense arrangement and exhibiting clear membrane boundary. Schisandrae Chinensis Fructus lignans inhibited the expression of SIRT 1 and FoxO3a in the hippocampus （P<0.05）. ConclusionTo sum up， Schisandrae Chinensis Fructus lignans may improve the behavior of schizophrenic mice by activating the SIRT1/FoxO3a signaling pathway to exert neuroprotective effects.

Objective: To identify the structure of the complementarity determining region (CDRs), the V(D)J rearrangement and somatic hypermutational characteristics of the heavy and light chains of a red blood cell blood group-specific monoclonal antibody. Methods: The hybridoma cell line secreting human IgM κ monoclonal anti-P antibody was used as the research object. Total RNA was extracted from cultured monoclonal cell line, and cDNA was obtained by reverse transcription PCR (RT-PCR) using random hexamers primers. It was then amplified and sequenced using primers specific for variable regions of the immunoglobulin heavy and light chains encoding the anti-P antibody. The sequences were aligned against the NCBI database using online Immunoglobulin BLAST (Ig-BLAST) tool. Results: The study determined the structure of the CDRs and framework regions (FRs) of the variable regions of human monoclonal anti-P immunoglobulin, as well as the characteristics of V(D)J rearrangement. Moreover, the closest VH, VD, and VJ germline alleles for the heavy chain and VL and VJ germline alleles for the light chain were also identified. The IgH gene rearrangment pattern of the monoclonal anti-P was IGHV6-1 * 01—IGHD5-18 02—IGHJ4 02 and IgL gene was IGκV1-12 01—IGκJ3 01. Nine base mutations occurred within the germline gene IGHV6-1 01 in variable region of heavy chain, whereas 5 base mutations were found in the germline gene IGκV1-12 01 in variable region of light chain, respectively. Conclusion: This study characterized the CDR structure in monoclonal antibody cell line targeting the high-frequency red blood cell P antigen, and provided a foundation for the construction of recombinant antibody expressing plasmids and transfomation of the immunoglobulin type.

Objective: To construct gene recombinant expression plasmids of human anti-P antibody with IgG1 and kappa chain based on the human hybridoma cell line. Methods: Starting from the specific RT-PCR products encoding the variable regions of the IgH chain and IgL chain of the anti-P monoclonal cell line, appropriate restriction enzyme digestion sites were introduced at both ends of the VH and VL fragments through nested PCR. The plasmids carrying the antibody constant region and the nested PCR products of VH and VL were ligated by the action of T4 ligase and subsequently transferred into competent E. coli DH5ɑ, and positive clones were selected in the antibiotic resistant LB medium. After sequence confirmation, recombinant plasmids DNA were transfected into HEK293T cells, and the recombinant antibody obtained in the culture supernatant. The characteristics of recombinant expression antibodies were determined by using rapid antibody isotying kit, serological agglutination tests and flow cytometry. Results: Recombinant gene expression vectors, pFUSEss-CHIg-hG1+VH, pFUSE2ss-CLIg-hκ+VL, were successfully constructed. Human IgG1 kappa light chain antibodies were detected in the supernatant of HEK293T cells transient transfected with recombinant plasmids. After the supernatant was ultra-filtered and concentrated, it could cause agglutination reactions with P antigen-positive red blood cells. The mean fluorescence intensity (MFI) of the reaction between recombinant antibodies and antigen-positive red blood cells in flow cytometry experiments was higher than that of antigen-negative red blood cells. Conclusion: The experimental study on the conversion of red blood group antibody types by genetic engineering technology represents a beneficial exploration towards establishing a feasible technical route for the development of genetic recombination and modification of antibodies reagent.

Objective To generate and characterize natural killer cell (NK cell)-conditional Cd226 gene knockout mice using Cre-loxP technology, and to explore the role of CD226 on NK cells in alleviating intestinal inflammation in a murine model of ulcerative colitis (UC). Methods NK cell-conditional Cd226 gene knockout mice were generated by crossing loxP-flanked Cd226 mice with Ncr1-Cre mice via the Cre-loxP system. Polymerase chain reaction (PCR) and agarose gel electrophoresis were used for genotyping. A UC model was established by dextran sulfate sodium (DSS) induction. Flow cytometry was performed to analyze CD226 expression levels on NK cells and the infiltration of related immune cells in colon tissues. Hematoxylin-eosin (HE) staining was performed to assess the degree of colonic inflammation. Results DNA gel electrophoresis and flow cytometry confirmed the successful generation of NK cell-specific Cd226 knockout mice. After conditional knockout of Cd226 in NK cells, inflammation in the UC mouse model was alleviated. Flow cytometry results showed a reduced proportion of NK cells in peripheral blood and the colon lamina propria, while HE staining demonstrated attenuated inflammatory responses. Conclusion Specific knockout of Cd226 in NK cells mitigates intestinal inflammation in UC mice by reducing NK cell numbers and inhibiting their pro-inflammatory functions.
Animals ; Colitis, Ulcerative/pathology* ; Killer Cells, Natural/metabolism* ; Mice, Knockout ; T Lineage-Specific Activation Antigen 1 ; Antigens, Differentiation, T-Lymphocyte/genetics* ; Mice ; Disease Models, Animal ; Mice, Inbred C57BL ; Male

Objective To observe the effects of Yixintai on lipid metabolism and the protein expressions of CPT-1 and CD36 in rats with heart failure;To explore the mechanism of its treatment of heart failure.Methods 106 out of 120 SD rats were selected to establish the heart failure model induced by myocardial infarction by ligating the left anterior descending coronary artery,and 14 rats were selected as the sham-operation group.The successful model rats were randomly divided into model group,trimetazidine group and Yixintai low-,medium-and high-dosage groups,with 14 rats in each group.The administration group was given corresponding drugs by gavage once a day for 4 weeks.LVEF,LVFS,LVIDd and LVIDs were measured by color doppler ultrasonography,the contents of ANP,BNP,LA and FFA in serum were detected by ELISA,the contents of TG,TC,LDL and HDL were detected by automatic biochemical analyzer,HE and Masson staining were used to observe the morphology of myocardial tissue,the expressions of CPT-1 and CD36 protein in myocardial tissue were detected by Western blot.Results Compared with the sham-operation group,LVEF and LVFS in the model group decreased(P<0.05),the LVIDs and LVIDd increased(P<0.05);the contents of serum ANP,BNP,LA,FFA,TG,TC and LDL increased(P<0.05),while the content of HDL decreased(P<0.05),with myocardial edema,irregular arrangement of myocardial fibers,increased inflammatory cell infiltration and collagen fiber deposition;the protein expressions of CPT-1 and CD36 in myocardial tissue decreased(P<0.05).Compared with the model group,the LVEF and LVFS in Yixintai each dosage groups and trimetazidine group increased(P<0.05),LVIDs and LVIDd decreased(P<0.05);the contents of ANP,BNP,LA,FFA,TG,TC and LDL in serum of Yixintai medium-and high-dosage groups and trimetazidine group decreased(P<0.05),the content of HDL increased(P<0.05);myocardial edema was improved,inflammatory cell infiltration was reduced,collagen fiber deposition was reduced,and the protein expressions of CPT-1 and CD36 in myocardial tissue increased(P<0.05).Conclusion Yixintai may improve myocardial energy metabolism and treat heart failure by increasing the expression of CPT-1 and CD36 protein in myocardial tissue and promoting fatty acid β oxidation.

BACKGROUND:Gradient artificial bone repair scaffolds can mimic unique anatomical features in musculoskeletal tissues,showing great potential for repairing injured musculoskeletal tissues. OBJECTIVE:To review the latest research advances in gradient artificial bone repair scaffolds for tissue engineering in the musculoskeletal system and describe their advantages and fabrication strategies. METHODS:The first author of the article searched the Web of Science and PubMed databases for articles published from 2000 to 2023 with search terms"gradient,bone regeneration,scaffold".Finally,76 papers were analyzed and summarized after the screening. RESULTS AND CONCLUSION:(1)As an important means of efficient and high-quality repair of skeletal system tissues,gradient artificial bone repair scaffolds are currently designed bionically for the natural gradient characteristics of bone tissue,bone-cartilage,and tendon-bone tissue.These scaffolds can mimic the extracellular matrix of native tissues to a certain extent in terms of structure and composition,thus promoting cell adhesion,migration,proliferation,differentiation,and regenerative recovery of damaged tissues to their native state.(2)Advanced manufacturing technology provides more possibilities for gradient artificial bone repair scaffold preparation:Gradient electrospun fiber scaffolds constructed by spatially differentiated fiber arrangement and loading of biologically active substances have been developed;gradient 3D printed scaffolds fabricated by layered stacking,graded porosity,and bio-3D printing technology;gradient hydrogel scaffolds fabricated by in-situ layered injections,simple layer-by-layer stacking,and freeze-drying method;and in addition,there are also scaffolds made by other modalities or multi-method coupling.These scaffolds have demonstrated good biocompatibility in vitro experiments,were able to accelerate tissue regeneration in small animal tests,and were observed to have significantly improved histological structure.(3)The currently developed gradient artificial bone repair scaffolds have problems such as mismatch of gradient scales,unclear material-tissue interactions,and side effects caused by degradation products,which need to be further optimized by combining the strengths of related disciplines and clinical needs in the future.

BACKGROUND:There are still great controversies in the etiology,clinical manifestations,diagnosis and treatment of pigmented villonodular synovitis.Bibliometric and visualization studies on pigmented villonodular synovitis can clarify the research development context and point out the direction for future research.OBJECTIVE:To analyze the global research status,hotspot,and trend of pigmented villonodular synovitis.METHODS:All publications related to pigmented villonodular synovitis from 1995 to 2023 were retrieved from Web of Science and CNKI.Citespace and bibliometrics were used to analyze the clustering,co-occurrence,and emergent words of all articles.The Web of Science database adopts subject headings plus free words for retrieval,while the CNKI database retrieves through subject headings.Finally,986 English articles and 599 Chinese articles were included.RESULTS AND CONCLUSION:(1)The United States has an absolute leading position in research in this field,ranking first in the number of published papers,H index,and cited times.China ranks the 4th in the total volume of published articles and 12th in the H index.The quality of published articles and international cooperation still need to be improved.(2)Cluster analysis of pigmented villonodular synovitis studies showed that the top five clusters were radiotherapy,soft tissue sarcoma,rheumatoid arthritis,magnetic resonance imaging,and diagnosis.(3)The key words that continued to emerge until 2023 were colony-stimulating factor 1,giant cell tumor of tendon sheath,case report,chromosome translocation,radiotherapy,expression,and kinase.(4)Based on keyword analysis and co-citation analysis,it is found that the research on the clinical characteristics of pigmented villonodular synovitis,the development of new colony-stimulating factor 1 inhibitors,and the application of colony-stimulating factor 1 inhibitors in the treatment process are current research hotspots.(5)Combining thematic evolution with the analysis of current research hotspots,based on clarifying the etiology,pathogenesis,and clinical characteristics of pigmented villonodular synovitis,improving the diagnostic accuracy of pigmented villonodular synovitis,enhancing the precision of treatment,and reducing the recurrence rate after treatment will be key issues that require focus in the future.

Objective To analyze the temporal trends of gout disezse burden in China from 1990 to 2021,and construct an age-period-cohort(APC)model to explore the independent effects of age,period,and birth cohort on epidemiological indicators,and predict the future burden of gout disease in China from 2022 to 2035.Methods Data on gout disease burden in China during 1990-2021 were extracted from the Global Burden of Disease(GBD)2021 database.Joinpoint regression analysis was used to assess temporal trends.The APC model was applied to evaluate the age,period and cohort effects on prevalence risk and disability-adjusted life years(DALYs).A Bayesian age-period-cohort(BAPC)model was employed to project the age-standardized prevalence rate(ASPR)and age-standardized DALY rate(ASDR)of gout in China from 2022 to 2035.Results From 1990 to 2021,the incidence,prevalence and DALYs of gout in China all increased substantially,with overall rising trends in the age-standardized incidence rate(ASIR),ASPR and ASDR.Compared with 1990,the incidence,prevalence and DALYs in 2021 increased by 160.45%,181.12%,and 175.93%,respectively,while their age-standardized rates increased by 23.74%,26.48%and 25.89%.Joinpoint regression analysis revealed that average annual percentage changes(AAPCs)of 0.73%for ASIR,0.82%for ASPR and 0.80%for ASDR during 1990-2021.In 2021,the number of cases and DALYs reached their peaks in males aged 55-59 years and females aged 65-69 years.Both prevalence and DALY rates increased steadily with age,with marked rises starting at age 30 in men and age 40 in women.Overall,males showed higher prevalence,DALYs and corresponding rates than those of females across all age groups.APC model results indicated that the age effect,period effect and cohort effects on prevalence and DALY rates presented an overall upward tread.Decomposition analysis showed that population aging contributed the most to the increase in incidence and DALYs from 1990 to 2021.BAPC projections suggested that by 2035,the ASPR and ASDR of gout in China reached 890.50 per 100,000 and 27.26 per 100,000,respectively.Conclusion The ASPR and ASDR of gout in China are projected to continue increasing from 2022 to 2035.Targeted public health strategies for high-risk populations are urgently needed to reduce the growing burden of gout.

BACKGROUND:There are still great controversies in the etiology,clinical manifestations,diagnosis and treatment of pigmented villonodular synovitis.Bibliometric and visualization studies on pigmented villonodular synovitis can clarify the research development context and point out the direction for future research.OBJECTIVE:To analyze the global research status,hotspot,and trend of pigmented villonodular synovitis.METHODS:All publications related to pigmented villonodular synovitis from 1995 to 2023 were retrieved from Web of Science and CNKI.Citespace and bibliometrics were used to analyze the clustering,co-occurrence,and emergent words of all articles.The Web of Science database adopts subject headings plus free words for retrieval,while the CNKI database retrieves through subject headings.Finally,986 English articles and 599 Chinese articles were included.RESULTS AND CONCLUSION:(1)The United States has an absolute leading position in research in this field,ranking first in the number of published papers,H index,and cited times.China ranks the 4th in the total volume of published articles and 12th in the H index.The quality of published articles and international cooperation still need to be improved.(2)Cluster analysis of pigmented villonodular synovitis studies showed that the top five clusters were radiotherapy,soft tissue sarcoma,rheumatoid arthritis,magnetic resonance imaging,and diagnosis.(3)The key words that continued to emerge until 2023 were colony-stimulating factor 1,giant cell tumor of tendon sheath,case report,chromosome translocation,radiotherapy,expression,and kinase.(4)Based on keyword analysis and co-citation analysis,it is found that the research on the clinical characteristics of pigmented villonodular synovitis,the development of new colony-stimulating factor 1 inhibitors,and the application of colony-stimulating factor 1 inhibitors in the treatment process are current research hotspots.(5)Combining thematic evolution with the analysis of current research hotspots,based on clarifying the etiology,pathogenesis,and clinical characteristics of pigmented villonodular synovitis,improving the diagnostic accuracy of pigmented villonodular synovitis,enhancing the precision of treatment,and reducing the recurrence rate after treatment will be key issues that require focus in the future.

Objective To analyze the temporal trends of gout disezse burden in China from 1990 to 2021,and construct an age-period-cohort(APC)model to explore the independent effects of age,period,and birth cohort on epidemiological indicators,and predict the future burden of gout disease in China from 2022 to 2035.Methods Data on gout disease burden in China during 1990-2021 were extracted from the Global Burden of Disease(GBD)2021 database.Joinpoint regression analysis was used to assess temporal trends.The APC model was applied to evaluate the age,period and cohort effects on prevalence risk and disability-adjusted life years(DALYs).A Bayesian age-period-cohort(BAPC)model was employed to project the age-standardized prevalence rate(ASPR)and age-standardized DALY rate(ASDR)of gout in China from 2022 to 2035.Results From 1990 to 2021,the incidence,prevalence and DALYs of gout in China all increased substantially,with overall rising trends in the age-standardized incidence rate(ASIR),ASPR and ASDR.Compared with 1990,the incidence,prevalence and DALYs in 2021 increased by 160.45%,181.12%,and 175.93%,respectively,while their age-standardized rates increased by 23.74%,26.48%and 25.89%.Joinpoint regression analysis revealed that average annual percentage changes(AAPCs)of 0.73%for ASIR,0.82%for ASPR and 0.80%for ASDR during 1990-2021.In 2021,the number of cases and DALYs reached their peaks in males aged 55-59 years and females aged 65-69 years.Both prevalence and DALY rates increased steadily with age,with marked rises starting at age 30 in men and age 40 in women.Overall,males showed higher prevalence,DALYs and corresponding rates than those of females across all age groups.APC model results indicated that the age effect,period effect and cohort effects on prevalence and DALY rates presented an overall upward tread.Decomposition analysis showed that population aging contributed the most to the increase in incidence and DALYs from 1990 to 2021.BAPC projections suggested that by 2035,the ASPR and ASDR of gout in China reached 890.50 per 100,000 and 27.26 per 100,000,respectively.Conclusion The ASPR and ASDR of gout in China are projected to continue increasing from 2022 to 2035.Targeted public health strategies for high-risk populations are urgently needed to reduce the growing burden of gout.