Aim To express and purify recombinant hCGH-CTP fusion protein in high-density suspension culture of Chinese hamster ovary cells (CHO-S), and to verify the lipid accumulation effect of rhCGH-CTP on 3T3-L1 mature adipocytes. Methods The recombinant protein expression vector (pcDNA3. 1-rhCGH-CTP) was constructed, achieved by fusing the human glycoprotein hormone beta 5/alpha 2 cDNA with CTP Linker. The expression plasmid was transiently transfected into the suspended CHO-S to express rhCGH-CTP protein and then purified, and the protein biological activity was verified. Intervention with 3T3-L1 mature adipocyte cells for 24 h was performed to detect the changes of intracellular triglyceride (TG) level. Results Western blot results showed that rhCGH-CTP protein was successfully expressed in CHO-S cells, and the yield was up to 715. 4 mg • L~ . The secreted protein was purified by AKTA pure system with higher purity that was up to 90% as identified by SDS-PAGE. In addition, the intracellular cAMP content of mature adipocytes with high expression of TSHR gene significantly increased after intervention with different concentrations of rhCGH-CTP protein by ELISA kit, indicating that rhCGH-CTP protein had biological activity. Oil red 0 staining showed that compared with the control group, the lipid content of mature adipocytes in the intervention groups with different concentrations of rhCGH-CTP protein significantly decreased (P < 0. 05) . Conclusions The rhCGH-CTP protein has been successfully expressed and purified with biological activity, and effectively reduce TG. This research provides an important theoretical basis for further revealing the physiological role of CGH protein and its potential application in clinical practice.

Humans ; Consensus ; Immunoglobulin Light-chain Amyloidosis/therapy* ; Heart ; Immunoglobulin Light Chains ; China

ObjectiveProtein arginine methyltransferases (PRMTs) play pivotal roles in numerous cellular biological processes. However, the precise regulatory effects of PRMTs on the fate determination of mesenchymal stromal/stem cells (MSCs) remain elusive. Our previous studies have shed light on the regulatory role and molecular mechanism of PRMT5 in MSC osteogenic differentiation. This study aims to clarify the role and corresponding regulatory mechanism of PRMT7 during the adipogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Methods(1) Human bone marrow-derived mesenchymal stem cells (hBMSCs) were cultured in a medium that induces adipogenesis. We used qRT-PCR and Western blot to monitor changes in PRMT7 expression during adipogenic differentiation. (2) We created a cell line with PRMT7 knocked down and assessed changes in PRMT7 expression and adipogenic capacity using Oil Red O staining, qRT-PCR and Western blot. (3) We implanted hBMSCs cell lines mixed with a collagen membrane subcutaneously into nude mice and performed Oil Red O staining to observe ectopic lipogenesis in vivo. (4) A cell line overexpressing PRMT7 was generated, and we examined changes in PRMT7 expression using qRT-PCR and Western blot. We also performed Oil Red O staining and quantitative analysis after inducing the cells in lipogenic medium. Additionally, we assessed changes in PPARγ expression. (5) We investigated changes in insulin-like growth factor 1 (IGF-1) expression in both PRMT7 knockdown and overexpressing cell lines using qRT-PCR and Western blot, to understand PRMT7’s regulatory effect on IGF-1 expression. siIGF-1 was transfected into the PRMT7 knockdown cell line to inhibit IGF-1 expression, and knockdown efficiency was confirmed. Then, we induced cells from the control and knockdown groups transfected with siIGF-1 in lipogenic medium and performed Oil Red O staining and quantitative analysis. Finally, we assessed PPARγ expression to explore IGF-1’s involvement in PRMT7’s regulation of adipogenic differentiation in hBMSCs. Results(1) During the adipogenesis process of hBMSCs, the expression level of PRMT7 was significantly reduced (P<0.01). (2) The adipogenic differentiation ability of PRMT7 knockdown group was significantly stronger than that of control group (P<0.001). (3) The ectopic adipogenic differentiation ability of PRMT7 knockdown group was significantly stronger than that of control group. (4) The adipogenic differentiation ability of the PRMT7 overexpression group was significantly weaker than that of the control group (P<0.01). (5) The expression level of IGF-1 increased after PRMT7 knockdown (P<0.000 1). The expression level of IGF-1 decreased after PRMT7 overexpression (P<0.000 1), indicating that PRMT7 regulates the expression of IGF-1. After siIGF-1 transfection, the expression level of IGF-1 in all cell lines decreased significantly (P<0.001). The ability of adipogenic differentiation of knockdown group transfected with siIGF-1 was significantly reduced (P<0.01), indicating that IGF-1 affects the regulation of PRMT7 on adipogenic differentiation of hBMSCs. ConclusionIn this investigation, our findings elucidate the inhibitory role of PRMT7 in the adipogenic differentiation of hBMSCs, as demonstrated through both in vitro cell-level experiments and in vivo subcutaneous transplantation experiments conducted in nude mice. Mechanistic exploration revealed that PRMT7’s regulatory effect on the adipogenic differentiation of hBMSCs operates via modulation of IGF-1 signaling pathway. These collective findings underscore PRMT7 as a potential therapeutic target for fatty metabolic disorders, thereby offering a novel avenue for leveraging PRMT7 and hBMSCs in the therapeutic landscape of relevant diseases.

Outer membrane vesicles (OMVs) are nanoscale vesicles secreted by Gram-negative bacteria. As a unique bacterial secretion, OMV secretion can help bacteria maintain the outer membrane stability or remove harmful substances. Studies have shown that local separation of outer membrane and peptidoglycan layers led by abnormalities in outer membrane protein function, abnormal structure or excessive accumulation of LPS, and erroneous accumulation of phospholipids in the outer leaflet, which can all lead to bacterial outer membrane protrusion and eventually bud formation of OMVs. Since OMVs are mainly composed of bacterial outer membrane and periplasmic components, the pathogen associated molecular patterns (PAMPs) on their surface can trigger strong immune responses. For example, OMVs can recruit and activate neutrophils, polarize macrophages to secrete large amounts of inflammatory factors. More importantly, OMVs can act as adjuvants to induce dendritic cell (DC) maturation to enhance adaptive immune response in the body. At the same time, OMVs are derived from bacteria, which make it easy to modify. The methods by genetic engineering and others can improve their tumor targeting, give them new functions, or reduce their immunotoxicity, which is conducive to their application in tumor therapy. OMVs not only induce apoptosis or pyroptosis of tumor cells, but also regulate the host immune system, which makes OMVs themselves have a certain killing effect on tumors. In addition, the tendency of neutrophils to inflammatory tumor sites and the formation of neutrophil extracellular traps enable OMVs to target tumor sites, and the suitable size and the characteristic that they are easily taken up by DCs give OMVs a certain lymphatic targeting ability. Therefore, OMVs are often employed as excellent drug or vaccine carriers in tumor therapy. This review mainly discusses the biological mechanism of OMVs, the regulatory effects of OMVs on immune cells, the functional modification strategies of OMVs, and their research progress in tumor therapy.

AIM To evaluate the quality of Beidougen Formula Granules.METHODS Fifteen batches of standard decoctions and three batches of formula granules were prepared,after which paste rate and contents,transfer rates of magnoflorine,daurisoline,dauricine were determined.HPLC specific chromatograms were established,and cluster analysis was adopted in chemical pattern recognition.RESULTS For three batches of formula granules,the paste rates were 15.1%-16.6%,the contents of magnoflorine,daurisoline,dauricine were 18.93-19.39,9.42-9.60,6.79-6.85 mg/g with the transfer rates of 34.42%-35.25%,43.81%-44.65%,27.27%-27.51%from decoction pieces to formula granules,respectively,and there were seven characteristic peaks in the specific chromatograms with the similarities of more than 0.95,which demonstrated good consistence with those of standard decoctions and accorded with related limit requirements.Fifteen batches of standard decoctions were clustered into two types,and the medicinal materials produced from Jilin,Hebei,Shangdong could be used for the preparation of formula granules.CONCLUSION This reasonable and reliable method can provide references for the quality control and clinical application of Beidougen Formula Granules.

Objective To investigate the effects of long non-coding RNA(lncRNA)SNHG15 on lipopolysaccharide(LPS)-induced injury of human alveolar epithelial cells and the underlying molecular mechanism.Methods A549 cells were treated with LPS to construct a neonatal acute respiratory distress syndrome(NRDS)cell model.A549 cells were divided into Control group,LPS group,LPS+si-NC group,LPS+si-lncRNA SNHG15 group,LPS+miR-NC group,LPS+miR-942-5p group,LPS+si-lncRNA SNHG15+anti-miR-NC group and LPS+si-lncRNA SNHG15+anti-miR-942-5p group.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression levels of lncRNA SNHG15 and miR-942-5p.Flow cytometry and Western blot were conducted to detect cell apoptosis.ELISA was used to detect the expression levels of interleukin-6(IL-6),in-terleukin-1β(IL-1β),and tumor necrosis factor-α(TNF-α).Dual-luciferase reporter assay was used to identify the targeting rela-tionship between lncRNA SNHG15 and miR-942-5p.Results Compared with the Control group,lncRNA SNHG15 expression,apoptosis rate,and levels of Bax,IL-6,IL-1β and TNF-α were increased in the LPS group,while miR-942-5p expression and Bcl-2 protein level were decreased(all P＜0.05).After knockdown of lncRNA SNHG15 or overexpression of miR-942-5p,cell apop-tosis rate and levels of Bax,IL-6,IL-1β and TNF-α were decreased,while Bcl-2 level was increased(all P＜0.05).lncRNA SNHG15 targeted miR-942-5p,and downregulation of miR-942-5p reversed the effect of lncRNA SNHG15 knockdown on LPS-induced A549 cell injury(all P＜0.05).Conclusion Silencing of lncRNA SNHG15 alleviates LPS-induced A549 cell injury by upregulation of miR-942-5p.

Objective To investigate the efficacy and safety of exchange transfusion in neonates with severe pertussis. Methods A retrospective analysis was performed for the medical data of five neonates with severe pertussis who underwent exchange transfusion in the Department of Neonatology,Hunan Children's Hospital,from August 2019 to March 2024. The clinical characteristics of the patients were summarized,and the efficacy and adverse reactions of exchange transfusion were analyzed. Results All five neonates had the symptoms of hypoxemia,recurrent apnea,and heart failure and required invasive mechanical ventilation. Two cases of pulmonary hypertension were observed,one of which was complicated by decompensated shock. Before exchange transfusion,the five children had a median leukocyte count of 82.60×109/L,a median absolute lymphocyte count of 28.20×109/L,and a median absolute neutrophil count of 43.10×109/L,and reexamination at 4 hours after exchange transfusion showed that these values decreased to 28.40×109/L,7.60×109/L,and 15.40×109/L,respectively. The four children who underwent exchange transfusion in the early stage of cardiopulmonary failure showed varying degrees of improvement in oxygenation and a reduction in the partial pressure of carbon dioxide,and they were discharged after improvement;the one child who underwent exchange transfusion in the late stage of cardiopulmonary failure ultimately died. No child experienced severe adverse reactions related to exchange transfusion. Conclusions For neonates with severe pertussis,initiating exchange transfusion in the early stages of cardiopulmonary failure can effectively reduce leukocyte levels,potentially improve survival rates,and is relatively safe.

Objective To investigate the clinical effect of orthopedic robot combined with Starr pelvic reduction frame in the treatment of Tile type C pelvic ring fracture.Methods From October 2019 to May 2021,14 patients with type C pelvic ring fracture were treated with robotic combined with Starr pelvic reduction frame,including 9 males and 5 females.The age ranged from 33 to 69 years.All the 14 patients had fresh closed fractures without femur,tibia and fibula fracture.Surgery was complet-ed from 4 to 7 d after hospital admission.During the operation,the X-ray carbon bed was used,the pelvic ring was reduced by Starr pelvis reduction frame,and pelvic ring fracture was treated by orthopedic robot.Operation time,bleeding volume,fluo-roscopy times of single screw placement,fracture reduction quality,affected limb function and complications were observed.Radiological reduction was evaluated using Matta scoring standard,and clinical efficacy was evaluated by Majeed pelvic func-tion scoring system at the final follow-up.Results All of 14 patients successfully completed the operation,the operation time was 84 to 141 min,the bleeding volume was 20 to 50 ml,and the fluoroscopy times of single screw insertion was 4 to 9 times.All of 14 patients were followed up for 12 to 24 months.The healing time was 3 to 7 months.No complications such as fracture of internal fixation,screw loosening,infection and nerve injury were found.According to the evaluation criteria of Matta imag-ing reduction,9 cases were excellent,4 cases were good,and 1 case was fair.At the final follow-up,Majeed pelvic function scoring system was used:10 cases were excellent,4 cases were good.Conclusion The treatment of type C pelvic ring fracture with robotic combined Starr pelvis reduction frame is simple,time-saving,less trauma,less complications and effective.

A 55-year-old male patient was admitted to the hospital due to "recurrent chest pain for 8 months, with worsening symptoms for 2 weeks". After admission, comprehensive relevant examinations led to the consideration of a giant chronic left ventricular pseudoaneurysm caused by myocardial infarction with non-obstructive coronary arteries. Surgical treatment was performed at our hospital. We discuss the diagnosis and treatment of this patient.

Objective To observe the effect of Buyang Huanwu Decoction on hippocampal tissue structure and blood perfusion in rats with focal cerebral ischemia using multi-modal MRI;To analyze the mechanism of Buyang Huanwu Decoction on hippocampal neuroplasticity combined with the expression changes of neuroplasticity proteins and glucose metabolism related transporters.Methods Focal cerebral ischemia rat model induced by right middle cerebral artery occlusion were established.The rats were divided into sham-operation group,sham-operation+Buyang Huanwu Decoction group,model group and model+Buyang Huanwu Decoction group.The rats in the sham-operation group and model group were given normal saline for 30 days,and those in the sham-operation+Buyang Huanwu Decoction group and model+Buyang Huanwu Decoction group were given Buyang Huanwu Decoction for 30 days.T2 mapping imaging was used to detect the changes in hippocampal tissue structure,the changes of cerebral blood perfusion in hippocampus were detected by arterial spin labeling imaging,Western blot was used to detect the protein expressions of SYN,GAP-43,glucose transporters and MCT.Results Compared with the sham-operation group,the T2 relaxation time of the right and left hippocampus in the model group rats significantly increased(P<0.01,P<0.001),the blood flow in the right hippocampus was significantly reduced(P<0.05),the protein expressions of SYN,GAP-43,MCT4 and MCT2 in the right hippocampus were significantly decreased(P<0.01,P<0.001),the protein expressions of GLUT1 and GLUT3 in bilateral hippocappal tissue significantly decreased(P<0.05,P<0.01,P<0.001).Compared with the model group,the T2 relaxation time in the right hippocampus of rats in model+Buyang Huanwu Decoction group was significantly reduced(P<0.05),the blood flow in the right hippocampus significantly increased(P<0.05),the protein expressions of SYN,GAP-43,GLUT1 and GLUT3 in bilateral hippocampal tissues significantly increased(P<0.01,P<0.001),and the protein expressions of MCT4 and MCT2 in right hippocampal tissue significantly increased(P<0.01,P<0.001).Conclusion Buyang Huanwu Decoction can alleviate hippocampal injury in rats with focal cerebral ischemia,which may be related to improving blood perfusion,up-regulating neuroplasticity-related proteins,promoting hippocampal axon regeneration and synaptic remodeling,regulating energy metabolism of nerve cells.