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.

Objective To investigate the clinical effect of dual incision and dual plate internal fixation for the treatment of tibial plateau fractures in the elderly and its impact on knee joint function.Methods Using retrospective analysis method,103 elderly patients with tibial plateau fractures admitted to Rugao Hospital affiliated to Nantong University from April 2015 to May 2021 were selected for clinical research.According to the surgical method,they were divided into two groups,52 patients in the study group were treated with double incision double plates,and 51 patients in the control group were treated with locking plate internal fixation.Perioperative indicators,knee function,knee stability Knee joint complications.Results The hospitalization time of the study group was(7.8±2.0)days,and that of the control group was(10.0±2.4)days,and the difference was statistically significant between the two groups(P＜0.05).The HSS scores at 3 months,6 months and 12 months were(66.9±5.4),(78.4±6.6)and(83.8±6.1)in the study group,and(64.2±6.1),(74.0±7.3)and(82.0±6.8)in the control group,respectively.There was statistical significance in HSS scores 3 months and 6 months after operation between the two groups(P＜0.05).The tibial migration distance 3 months after operation was(2.54±0.50)mm in the study group and(2.84±0.67)mm in the control group,and the difference was statistically significant(P＜0.05).The knee posterior inclination angles of the study group were(4.12±1.10)°,(5.03±0.96)° and(5.46±1.52)° at 3 months,6 months and 12 months after surgery,respectively,while those of the control group were(6.11±1.43)°,(6.67±1.54)° and(7.50±1.88)°,respectively.The difference was statistically significant(P＜0.05).The activity of the study group at 3 months and 6 months after surgery was(104.3±8.2)° and(117.4±7.6)°,respectively,while that of the control group was(96.8±8.9)° and(111.8±8.2)°,and the difference was statistically significant(P＜0.05).There were 3 cases of postoperative complications in the study group and 10 cases in the control group,and the difference was statistically significant(P＜0.05).Conclusion The clinical effect of dual incision and dual plate internal fixation in the treatment of tibial plateau fractures in the elderly is affirmative,which is more beneficial for promoting early recovery of knee function and maintaining stability of knee function than locking plate internal fixation.

Osteoarthritis(OA),characterized by cartilage degeneration,synovial inflammation,and subchondral bone remodeling,is among the most common musculoskeletal disorders globally in people over 60 years of age.The initiation and progression of OA involves the abnormal metabolism of chondrocytes as an important pathogenic process.Cartilage degeneration features mitochondrial dysfunction as one of the important causative factors of abnormal chondrocyte metabolism.Therefore,maintaining mitochondrial homeostasis is an important strategy to mitigate OA.Mitophagy is a vital process for autophagosomes to target,engulf,and remove damaged and dysfunctional mitochondria,thereby maintaining mitochondrial homeostasis.Cumulative studies have revealed a strong association between mitophagy and OA,suggesting that the regulation of mitophagy may be a novel therapeutic direction for OA.By reviewing the literature on mitophagy and OA published in recent years,this paper elaborates the potential mechanism of mitophagy regulating OA,thus providing a theoretical basis for studies related to mitophagy to develop new treatment options for OA.

The endoplasmic reticulum is a key site for protein production and quality control.More than one-third of proteins are synthesized and folded into the correct three-dimensional conformation in the endoplasmic reticulum.However,during protein folding,unfolded and/or misfolded proteins are prone to occur,which may lead to endoplasmic reticulum stress.Organisms can monitor the quality of the proteins produced by endoplasmic reticulum quality control(ERQC)and endoplasmic reticulum-associated degradation(ERAD),which maintain endoplasmic reticulum protein homeostasis by degrading abnormally folded proteins.The underlying mechanisms of protein folding and ERAD in mammals have not yet been fully explored.Therefore,this paper reviews the process and function of protein folding and ERAD in mammalian cells,in order to help clinicians better understand the mechanism of ERAD and to provide a scientific reference for the treatment of diseases caused by abnormal ERAD.

Aim To investigate the effect of recombi-nant glycoprotein hormone β5/α2(rCGH)on lipolysis in 3T3-L1 adipocytes,and explore the underlying mechanism.Methods 3T3-L1 preadipocytes were cultured and induced to differentiate into mature adipo-cytes,then treated with different concentrations of rCGH for 24 h in vitro.Cell viability of 3T3-L1 adipo-cytes was evaluated by CCK-8 assay,the levels of in-tracellular triglyceride(TG)and glycerol in the culture supernatant were measured by enzymatic method,and the changes of lipid droplets were observed by oil red O staining.The expression levels of HSL and ATGL lipo-lytic proteins in adipocytes were detected by Western blot.To carry out the intervention experiment with dif-ferent concentrations of rCGH with or without the PKA inhibitor,H89,on the mature 3T3-L1 adipocytes,the cultured cells were divided into the control group,H89 pre treatment group,1 μmol·L-1 rCGH group,and(1 μmol·L-1 rCGH+H89)combined intervention group.The contents of intracellular TG and free glycer-ol were measured by enzymatic method,and the ex-pression of CREB and lipolysis-related proteins was de-tected using Western blot.Results Different concen-trations of rCGH(0.25,0.5,1,and 2 μmol·L-1)had no significant effect on the cell viability of adipo-cytes(P＞0.05).Compared with the control group,the treatment with rCGH significantly decreased the size of lipid droplets and intracellular TG content,while significantly elevated glycerol concentration in cell supernatant.rCGH treatment also stimulated the protein expression of p-HSL,ATGL,and p-PKA.In addition,the addition of a PKA inhibitor,H89,atten-uated the effects of rCGH on free glycerol level,intra-cellular TG content,and the expression of p-HSL,p-PLIN1,and p-CREB.Conclusions rCGH enhances the lipolysis of 3T3-L1 adipocytes by up-regulating the activities of HSL,ATGL and PKA,promoting glycerol release,inhibiting TG synthesis and lipid accumula-tion,and its mechanism of action is related to the acti-vation of cAMP/PKA/CREB signaling pathway.

Aim To investigate the effects of Ginkgo biloba extract(GBE)on vascular endothelial dysfunc-tion induced by chronic intermittent hypoxia(CIH)in rats and its related mechanisms.Methods The CIH rat model was established,and 50 and 100 mg·kg-1 GBE was administered by intragastric administration.The systolic blood pressure(SBP)of the tail artery was detected in each group.HE staining was used to detect the morphology of aorta tissue.DAF-FM DA staining and nitric reductase assay were used to detect NO levels.ELISA was used to detect serum ET-1,TNF-α and IL-6 levels.DHE staining was used to de-tect reactive oxygen species(ROS)levels of aortic tis-sue.Kits were used to detect the serum levels of MDA,SOD and GSH-Px.Western blot was used to detect the levels of VCAM-1,ICAM-1,nucleus Nrf2,HO-1 and NQO1 of aortic tissue.Results GBE sig-nificantly decreased the levels of SBP,ET-1,ROS,MDA,VCAM-1,ICAM-1,TNF-α and IL-6,and sig-nificantly increased the levels of NO,SOD,GSH-Px,nuclear Nrf2,HO-1 and NQO1 in CIH rats.GBE sig-nificantly improved the histomorphology of aorta in CIH rats.Conclusions GBE can improve vascular endo-thelial dysfunction and reduce blood pressure in CIH model rats.The mechanism may be related to the acti-vation of Nrf2/ARE pathway and the inhibition of oxi-dative stress and inflammation by GBE.

Aim To investigate the effects of dagliflozin (DAPA) on atrial tachyarrhythmia (AT) in rats with right heart failure (RHF) due to pulmonary arterial hypertension (PAH) and the underlying mechanisms. Methods Sixty male SD rats were randomly divided into four groups: control group (CTL group), model group (MCT group), MCT + low-dose DAPA intervention group (MCT + LD group) and MCT + high-dose DAPA intervention group (MCT + HD group). After 35 days of continuous intervention, the model and cardiac function evaluation, atrial structural remodelling assessment, inflammatory factor detection, and in vivo cardiac electrophysiology experiments were completed. Results DAPA reduced menn pulmonaryarterial pressure (mPAP) and menn right ventricular pressure (mRVP) in the model rats (P <0.05), attenuated the inflammatory response (P < 0.05), reduced right atrial fibrosis (P <0.05), reduced AT induction rate (P < 0.05) and mean atrial tachyarrhythmia duration (MATD) (P < 0.05), the extent of which was more pronounced in the high-dose DAPA intervention group. Conclusions DAPA can reduce AT susceptibility in PAH-induced RHF rats, and the mechanisms may be related to the inhibition of systemic inflammation and anti-atrial fibrosis by DAPA.

Objective:To investigate the effect of miR-503 targeting CBX2 on drug resistance of breast cancer MDA-MB-231 cells and its potential mechanism.Methods:miR-con group, miR-503 group, si-con group, two groups of si-chromosome homologues (CBX), anti-miR-con group, anti-miR-503 group, miR-503+pcDNA group, miR-503+pcDNA-CBx2 group were set up. Real-time quantitative fluorescence polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-503 and CBX2 mRNA. Western blot was used to detect protein expression. Cell activity was detected by MTT assay. The targeted regulatory relationship was detected by double luciferase assay.Results:Compared with normal breast cells HBL-100 (1.02±0.09), the expression level of miR-503 in breast cancer cells MCF-7 (0.41±0.05), MDA-MB-231 (0.25±0.03) and BT474 (0.35±0.04) was significantly decreased. The expression levels of CBX2 mRNA in MCF-7, MDA-MB-231 and BT474 cells were (4.02±0.35), (4.62±0.36) and (3.47±0.33), respectively. The expression levels of CBX2 protein in MCF-7, MDA-MB-231 and BT474 cells were (0.64±0.07), (0.74±0.05) and (0.68±0.06), respectively. The mRNA and protein contents of CBX2 in normal breast cells HPL-100 were (1.01±0.08) and (0.40±0.04), respectively, and the expression of CBX2 in breast cancer cells was significantly higher than that in normal breast cells ( P<0.05). Overexpression of miR-503 (3.64± 0.30) and silting of CBX2 inhibited proliferation, migration and invasion of MDA-MB-231 cells, and inhibited CBX2 (0.26±0.03), cyclin-dependent kinases, CDK) 4 (0.32± 0.03), Cyclin (CCN) D1 (0.58±0.03), matrix metalloproteinases (matrix metalloproteinases), MMP-2 (0.32±0.03) and MMP-9 (0.32±0.04) ( P<0.05). miR-503 targeted the expression of CBX2, and overexpression of CBX2 (0.75±0.03) could reverse the proliferation and drug resistance of miR-503 to MDA-MB-231 breast cancer cells. Conclusion:miR-503 may inhibit the proliferation, migration and invasion of breast cancer cells by down-regulating the expression of CBX2.

Objective This study aims to explore the formation of the dilemma of"giving care"for disabled elderly people in nursing homes from the perspective of active health.Methods Purposive sampling was used to select caregivers and disabled elderly people from a nursing home in Chongqing as research subjects from July 2022 to December 2022.One-on-one in-depth interviews were conducted and Colaizzi's 7-step thematic analysis method was employed to collect and analyze the interview data.Results 4 themes were identified:①the dominance of the"giving"care concept,including the cultural thoughts of filial piety,passive acceptance of care characteristics and consumer psychology regarding paid services;②insufficient"participation"care ability,including lack of knowledge regarding active health and a weakening of skills to promote participation;③the hindrance to"transformation"of care models,including objective limitations in terms of human resources and delays in adapting aging environments;and ④the decline in"utilization"of internal abilities,including excessive avoidance of potential risks,a heavy workload of care and poor quality of individual care.Conclusion The formation of the dilemma of"giving care"for disabled elderly people in nursing homes is affected by multiple factors such as social background,service system,supply resources,and management mode.Transforming disabled elderly individuals from a state of"passive giving care"to"active participation in their health"is an important measure to realize the concept of positive aging and healthy aging.

H 1-antihistamines are widely used in the treatment of various allergic diseases, but there are still many challenges in the safe and rational use of H 1-antihistamines in pediatrics, and there is a lack of guidance on the prescription review of H 1-antihistamines for children.In this paper, suggestions are put forward from the indications, dosage, route of administration, pathophysiological characteristics of children with individual difference and drug interactions, so as to provide reference for clinicians and pharmacists.