ObjectiveTo investigate the mechanism through which miR‑21 improves chronic renal fibrosis in mice via targeted modulation of the phosphatase and tensin homolog （PTEN）/protein kinase B （AKT）/mammalian target of rapamycin （mTOR） pathway. MethodsThirty‑two chronic kidney disease model mice were randomly divided into four groups （n=8 each group）： model group， miR‑21 overexpression group， miR‑21 inhibition group， and miR‑21 inhibition + MK‑2206 group. Eight healthy mice were included as the control group. The miR‑21 overexpression， miR‑21 inhibition， and miR‑21 inhibition + MK‑2206 groups received tail‑vein injections of lentivirus （50 μL， 1×10⁸ TU per mouse） once weekly for three weeks. The control and model groups were injected with an equal volume of empty vector （LV‑NC）. The miR‑21 inhibition + MK‑2206 group additionally received gavage of the AKT/mTOR pathway inhibitor MK‑2206 （480 mg/kg） once weekly for three weeks. The expressions of miR‑21， 24 h urinary protein， serum creatinine （Scr）， blood urea nitrogen （BUN）， and renal tissue levels of collagen Ⅰ， collagen Ⅲ， α‑smooth muscle actin （α‑SMA）， and PTEN protein， as well as p‑AKT/AKT and p‑mTOR/mTOR ratios， were compared among groups. HE staining was used to observe pathological changes in renal tissue， and Masson staining was used to observe the degree of renal fibrosis. A dual‑luciferase assay was performed to verify the targeting relationship between miR‑21 and PTEN. ResultsCompared with the model group， miR‑21 expression in renal tissue increased in the miR‑21 overexpression group （P<0.05） and decreased in the miR‑21 inhibition group （P<0.05）. Compared with the model group， the miR‑21 overexpression group showed increased 24 h urinary protein， Scr， BUN， and renal tissue expression of collagen Ⅰ， collagen Ⅲ， and α‑SMA （all P<0.05）， while these indicators decreased in the miR‑21 inhibition group （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group exhibited lower 24‑h urinary protein， Scr， BUN， and renal tissue expression of Collagen Ⅰ， Collagen Ⅲ， and α‑SMA （all P<0.05）. Compared with the model group， the miR‑21 overexpression group showed decreased PTEN protein expression （P<0.05） and increased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， while the miR‑21 inhibition group showed increased PTEN expression （P<0.05） and decreased p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）. Compared with the miR‑21 inhibition group， the miR‑21 inhibition + MK‑2206 group had lower p‑AKT/AKT and p‑mTOR/mTOR ratios （P<0.05）， with no significant difference in PTEN protein expression. HE and Masson staining showed normal kidney structure and almost no fibrosis in the control group. The model group exhibited glomerular enlargement， capillary loop adhesion， and focal fibrosis. The miR-21 overexpression group showed severe destruction of glomerular structure， accompanied by extensive fibrosis and renal tubular atrophy. The pathological changes and degree of fibrosis were alleviated in the miR-21 inhibition group. The miR-21 inhibition + MK-2206 group showed only mild pathological changes and mild fibrosis， with the interstitium being largely normal. Compared with PTEN-WT + NC mimics 1， the relative luciferase activity in the PTEN-WT + miR-21 mimics group decreased （P<0.001）. There was no statistically significant difference in relative luciferase activity between PTEN-WT + NC mimics group and PTEN-MUT + miR-21 mimics group. ConclusionmiR‑21 may improve renal function indicators and alleviate renal fibrosis in chronic kidney disease mice via targeted modulation of PTEN and subsequently inhibiting the AKT/mTOR pathway.

Objective:To explore the dynamic relationship between college students' online psychological needs satisfaction dominance(OPNSD)and phubbing.OPNSD refers to the tendency of individuals to fulfill their basic psychological needs primarily through online environments.Methods:A sample of 340 college students from a university in Beijing participated in two data collections(T1 and T2)at 3-month intervals.OPNSD was calculated as the difference between scores on the Need Satisfaction Perceived Online Scale(NSPOS)and the Basic Need Sat-isfaction in General Scale(BNSGS).Phubbing and its 4 dimensions(self-isolation,nomophobia,interpersonal con-flict,and problem acknowledgment)were measured with the Generic Scale of Phubbing(GSP).A cross-lagged panel model was used to explore the relationship between the online psychological needs satisfaction dominance and phubbing.Results:Significant simultaneous(r=0.60,P＜0.001)and successive temporal(r=0.56,P＜0.001)correlations were observed between the OPNSD score and GSP score.T1 OPNSD score was positively associated with T2 GSP score in the dimensions of self-isolation,nomophobia,and interpersonal conflict(β=0.14,0.10,0.11;P＜0.001 or P＜0.05),but showed no association with the problem acknowledgement dimension score(β=0.07,P＞0.05).However,the four dimensions of T1 GSP score were not associated with T2 OPNSD score(Ps＞0.05).Conclusion:This study suggests that college students' online psychological needs satisfaction dominance positively predicts phubbing,highlighting the critical influence of online environments on mobile phone behaviors.

AIM:The aim of this study was to explore the effects of Yiqi-Yangyin-Quyu prescription(YP)on skeletal muscle injury and related metabolic disorders in mice with Sj?gren syndrome(SS),and to clarify the role of hy-droxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit beta(Hadhb)in mediating the effect of YP on skeletal muscle in SS.METHODS:The SS mice underwent YP treatment for 8 weeks.The morphological changes of the submandibular gland and muscle tissue were examined using hematoxylin-eosin staining.The mitochondrial status in mus-cle tissue was assessed through transmission electron microscopy.Additionally,combined transcriptome and proteome se-quencing was conducted on skeletal muscle samples.The omics sequencing results were validated by RT-qPCR.Immuno-fluorescence was used to confirm the levels of key proteins involved in the P53/peroxisome proliferator-activated receptor gamma(PPARG)signaling pathway.Immunohistochemistry and Western blot were employed to determine the levels of Hadhb key targets.RESULTS:Combined transcriptome and proteome analysis identified 1 523 differentially expressed genes(DEGs)and 182 differentially expressed proteins(DEPs)between the muscle tissue of SS mice(model group)and that of control animals(ICR group),12 of which showed co-differential expression at both transcriptomic and proteomic levels.Compared with model group,1 232 genes and 432 proteins were found to be differentially expressed in the muscle tissue of the mice in YP group.Among these,23 exhibited co-differential expression at both mRNA and protein levels.Gene Ontology(GO)analysis showed that the DEGs and DEPs between ICR and model groups were mainly involved in ener-gy metabolism and fatty acid oxidation,while the DEGs and DEPs between YP and model groups were primarily associated with sarcomere tissue and actin structure.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis indi-cated that the DEGs and DEPs between ICR and model groups and between model and YP groups were enriched in the com-plement and coagulation cascade and lipid and pyruvate metabolism.The RT-qPCR validation results were consistent with those of the transcriptome analysis.Furthermore,the protein expression of the tumor suppressor P53 was significantly de-creased in YP group compared with model group,whereas that of PPARG was significantly increased.Western blot analy-sis showed that compared with ICR group,Hadhb protein expression was significantly decreased in model group,whereas the opposite trend was detected in YP group.CONCLUSION:The SS-related skeletal muscle damage is closely related to amino acid metabolism disorder and fatty acid degradation.Treatment with YP modulates innate immune defenses,lipid metabolism and energy metabolism in SS,and Hadhb is the key target of YP in SS-related skeletal muscle.

Objective:To explore the dynamic relationship between college students' online psychological needs satisfaction dominance(OPNSD)and phubbing.OPNSD refers to the tendency of individuals to fulfill their basic psychological needs primarily through online environments.Methods:A sample of 340 college students from a university in Beijing participated in two data collections(T1 and T2)at 3-month intervals.OPNSD was calculated as the difference between scores on the Need Satisfaction Perceived Online Scale(NSPOS)and the Basic Need Sat-isfaction in General Scale(BNSGS).Phubbing and its 4 dimensions(self-isolation,nomophobia,interpersonal con-flict,and problem acknowledgment)were measured with the Generic Scale of Phubbing(GSP).A cross-lagged panel model was used to explore the relationship between the online psychological needs satisfaction dominance and phubbing.Results:Significant simultaneous(r=0.60,P＜0.001)and successive temporal(r=0.56,P＜0.001)correlations were observed between the OPNSD score and GSP score.T1 OPNSD score was positively associated with T2 GSP score in the dimensions of self-isolation,nomophobia,and interpersonal conflict(β=0.14,0.10,0.11;P＜0.001 or P＜0.05),but showed no association with the problem acknowledgement dimension score(β=0.07,P＞0.05).However,the four dimensions of T1 GSP score were not associated with T2 OPNSD score(Ps＞0.05).Conclusion:This study suggests that college students' online psychological needs satisfaction dominance positively predicts phubbing,highlighting the critical influence of online environments on mobile phone behaviors.

AIM:The aim of this study was to explore the effects of Yiqi-Yangyin-Quyu prescription(YP)on skeletal muscle injury and related metabolic disorders in mice with Sj?gren syndrome(SS),and to clarify the role of hy-droxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit beta(Hadhb)in mediating the effect of YP on skeletal muscle in SS.METHODS:The SS mice underwent YP treatment for 8 weeks.The morphological changes of the submandibular gland and muscle tissue were examined using hematoxylin-eosin staining.The mitochondrial status in mus-cle tissue was assessed through transmission electron microscopy.Additionally,combined transcriptome and proteome se-quencing was conducted on skeletal muscle samples.The omics sequencing results were validated by RT-qPCR.Immuno-fluorescence was used to confirm the levels of key proteins involved in the P53/peroxisome proliferator-activated receptor gamma(PPARG)signaling pathway.Immunohistochemistry and Western blot were employed to determine the levels of Hadhb key targets.RESULTS:Combined transcriptome and proteome analysis identified 1 523 differentially expressed genes(DEGs)and 182 differentially expressed proteins(DEPs)between the muscle tissue of SS mice(model group)and that of control animals(ICR group),12 of which showed co-differential expression at both transcriptomic and proteomic levels.Compared with model group,1 232 genes and 432 proteins were found to be differentially expressed in the muscle tissue of the mice in YP group.Among these,23 exhibited co-differential expression at both mRNA and protein levels.Gene Ontology(GO)analysis showed that the DEGs and DEPs between ICR and model groups were mainly involved in ener-gy metabolism and fatty acid oxidation,while the DEGs and DEPs between YP and model groups were primarily associated with sarcomere tissue and actin structure.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis indi-cated that the DEGs and DEPs between ICR and model groups and between model and YP groups were enriched in the com-plement and coagulation cascade and lipid and pyruvate metabolism.The RT-qPCR validation results were consistent with those of the transcriptome analysis.Furthermore,the protein expression of the tumor suppressor P53 was significantly de-creased in YP group compared with model group,whereas that of PPARG was significantly increased.Western blot analy-sis showed that compared with ICR group,Hadhb protein expression was significantly decreased in model group,whereas the opposite trend was detected in YP group.CONCLUSION:The SS-related skeletal muscle damage is closely related to amino acid metabolism disorder and fatty acid degradation.Treatment with YP modulates innate immune defenses,lipid metabolism and energy metabolism in SS,and Hadhb is the key target of YP in SS-related skeletal muscle.

BACKGROUND:Knee osteoarthritis is a common clinical degenerative joint disease characterized by chronic inflammation and oxidative stress.Resveratrol has anti-inflammatory and anti-oxidative stress biological effects,and therefore it can be used symptomatically and expected to provide a new strategy for the treatment of knee osteoarthritis. OBJECTIVE:To investigate the therapeutic effect and mechanism of resveratrol on knee osteoarthritis in rats through the silence information regulator 1(SIRT1)/forkhead transcription factor O1(FOXO1)pathway. METHODS:Forty Sprague-Dawley rats were randomly divided into control group,model group,low-dose resveratrol group,and high-dose resveratrol group,with 10 rats in each group.Knee osteoarthritis models were established in the model group,low-dose resveratrol group,and high-dose resveratrol group.A mixture of 4%papain solution and 0.3 mol/L cysteine solution(1:1 for 0.5 hours;20 μL)was injected at 1,4,and 7 days after modeling.Rats in the low-dose and high-dose resveratrol groups were injected with 25 and 100 mg/kg resveratrol through the articular cavity at 1 day after successful modeling,while those in the control and model groups were injected with equivalent volume of physiological saline through the articular cavity.After 28 days of treatment,the maximum knee joint activity was measured;the levels of oxidative stress indicators and inflammatory factors in the synovial fluid of the knee joint were analyzed by radioimmunoassay and ELISA;the content of collagen fibers in the knee joint was analyzed by safranin O-fast green staining;the degree of arthritic lesions was analyzed using the Mankin histological score;and the levels of SIRT1 and FOXO1 in the knee joint were detected by western blot assay. RESULTS AND CONCLUSION:Compared with the model group,the maximum knee flexion and extension angles of rats significantly increased in the low-dose and high-dose resveratrol groups,and were significantly higher in the high-dose group than the low-dose group(P<0.05).Compared with the model group,the levels of superoxide dismutase and glutathione peroxidase in the knee joint fluid of rats significantly increased in the low-dose and high-dose resveratrol groups.The level of malondialdehyde significantly decreased in both resveratrol groups,and the level in the high-dose resveratrol group was significantly better than that in the low-dose resveratrol group(P<0.05).Compared with the model group,the low-dose and high-dose resveratrol groups showed a significant decrease in the levels of interleukin 1β,interleukin 6 and tumor necrosis factor α in the knee joint fluid of rats,and the levels of these inflammatory factors were significantly lower in the high-dose resveratrol group than the low-dose resveratrol group(P<0.05).Compared with the model group,the content of collagen fibers in the knee joint was significantly increased in both resveratrol groups,and the high-dose resveratrol group showed a higher content of collagen fibers than the low-dose resveratrol group(P<0.05).Compared with the model group,the expression level of SIRT1 in the knee joints of rats significantly increased in both resveratrol groups,while the level of acetylated FOXO1 significantly decreased(P<0.05).The magnitude of changes was significantly better in the high-dose group than the low-dose group.To conclude,resveratrol significantly improves the levels of oxidative stress and inflammatory factors in the joint fluid of rats with knee osteoarthritis and alleviates arthritic symptoms in a dose-dependent manner,possibly through the SIRT1/FOXO1 pathway.

Objective:To explore the clinical effect and safety of intrinsic-nourishing exercise and oral Chinese medicine combined with conventional western medicine therapy in the treatment of perimenopause with insomnia.Methods:Prospective cohort study. A total of 60 perimenopause with insomnia visiting the Hebei Medical Qigong Hospital were enrolled as the research objects between June 2019 and June 2021. According to random number table method, they were divided into the control group and the observation group, 30 in each group. The control group was treated with oral estazolam tablets, while the observation group was treated with intrinsic-nourishing exercise combined with oral Chinese medicine on basis of the control group. All the patients were treated for 4 weeks as a course, and totally 2 courses. The levels of serum estradiol (E 2), FSH, and LH were detected by automatic chemiluminescence immunoassay analyzer. Pittsburgh sleep quality Index (PSQI) was used to evaluate sleep quality, and the quality of life was evaluated by the MOS 36-item Short Form Health Survey (SF-36). And the responsive rates, sleep quality, scores of TCM symptoms, and adverse reactions were compared before and after treatment. Results:The total response rate of observation group was significantly higher than that of the control group (90.0% vs. 66.7%; χ2=4.81, P<0.05). After treatment, PSQI scores of sleep quality, time to fall asleep, sleep duration, sleep efficiency, sleep disturbance, use of hypnotics, and daytime function in the observation group were significantly lower than those in the control group ( t=14.11, 12.49, 9.88, 13.54, 9.47, 14.11, 17.91, P<0.01). After treatment, the TCM symptom scores of insomnia with more dreams, waist and knee soreness, five upsets, fatigue and forgetfulness in the observation group were significantly lower than those in the control group ( t=9.51, 13.08, 16.17, 12.81, P<0.01). After treatment, the E 2 [(35.16±3.61) mmol/L vs. (31.06±3.12) mmol/L, t=4.71] in the observation group was significantly higher than that of the control group ( P<0.01), while the FSH [(69.61±6.04) U/L vs. (73.26±7.41) U/L, t=2.09], and LH [(32.21±3.35) U/L vs. (36.04±3.49) U/L, t=4.34] in the observation group were significantly lower than those in the control group ( P<0.05 or P<0.01). At 4 and 8 week after treatment, the SF-36 scores in the observation group were significantly higher than those in the control group ( t=6.30, 4.36, P<0.01). During treatment, 16.7% (5/30) adverse reaction happened in the observation group, while 10.0% (3/30) in the control group, but there was no statistical significant difference between two groups ( χ2=0.56， P=0.448). Conclusion:The intrinsic-nourishing exercise and oral Chinese medicine combined with conventional western medicine therapy can significantly improve clinical curative effect, improve sleep quality and TCM symptoms, regulate hormones and quality of life in perimenopause with insomnia.

Objective:To observe any effect of repeated magnetic stimulation of the spine on lower urinary tract function and the life quality of patients with neurogenic bladder after suprasacral spinal cord injury (SCI).Methods:Fifteen suprasacral SCI patients whose lower urinary tract function was not improving were enrolled. In the first 2 weeks, all received water drinking management and intermittent catheterization, while in the following 4 weeks they were additionally provided with daily transspinal magnetic stimulation at the level of the spinous process of the first lumbar vertebra five times a week. The stimulation frequency was 1Hz. The patients kept voiding diaries. Their urodynamics were quantified using neurogenic bladder symptom scoring (NBSS) and a life quality scale.Results:The frequency of catheterization and the average voided volume, as well as the maximum detrusor pressure during the storage phase, maximum bladder capacity, maximum urethral pressure during the voiding phase and voiding efficiency at the end of the sixth week were significantly different from those at the end of the second week and before the intervention. The average NBSS and life quality scores then showed significant differences from the earlier time points.Conclusion:Repeteitive transspinal magnetic stimulation of the spine can improve lower urinary tract functioning and the life quality of persons with neurogenic bladder after a suprasacral SCI.

Bromodomain containing protein 4 (BRD4), as an epigenetic reader, can specifically bind to the acetyl lysine residues of histones and has emerged as an attractive therapeutic target for various diseases, including cancer, cardiac remodeling and heart failure. Herein, we described the discovery of hit 5 bearing 4-phenylquinazoline skeleton through a high-throughput virtual screen using 2,003,400 compound library (enamine). Then, structure-activity relationship (SAR) study was performed and 47 new 4-phenylquinazoline derivatives toward BRD4 were further designed, synthesized and evaluated, using HTRF assay set up in our lab. Eventually, we identified compound C-34, which possessed better pharmacokinetic and physicochemical properties as well as lower cytotoxicity against NRCF and NRCM cells, compared to the positive control JQ1. Using computer-based molecular docking and cellular thermal shift assay, we further verified that C-34 could target BRD4 at molecular and cellular levels. Furthermore, treatment with C-34 effectively alleviated fibroblast activation in vitro and cardiac fibrosis in vivo, which was correlated with the decreased expression of BRD4 downstream target c-MYC as well as the depressed TGF-β1/Smad2/3 signaling pathway. Taken together, our findings indicate that novel BRD4 inhibitor C-34 tethering a 4-phenylquinazoline scaffold can serve as a lead compound for further development to treat fibrotic cardiovascular disease.

Objective:To establish an in vitro growth competition assay using HIV-1 CRF07_BC infectious clone containing the mouse Thy1.1 gene or Thy1.2 gene. Methods:The mouse Thy1.1 gene and Thy1.2 gene were used to replace the 218 bases of Nef region of the CRF07_BC infectious clone (pXJDC6291-13) and construct the CRF07_BC infectious clones BCEA1 and BCEA2 plasmid. BCEA1 and BCEA2 plasmids were transfected into 293T to obtain the virus and the viral titration was measured. PM1 cells were co-infected with an equal virus. On the 3 rd to 6 th day of infection the cells were collected and labeled with specific antibodies Thy1.1 and Thy1.2. The viral fitness was detected by flow cytometry. Through "TFitness" (http: //bis.urmc.rochester.edu/vFitness) the relative viral fitness was calculated. The influence K103 N and K166R on the relative fitness of HIV-1 CRF07_BC were observed. Results:The relative fitness (1+ s) of CRF07_BC subtypes BCEA1 and BCEA2 viruses were 1.06±0.23693, which was not statistically significant. An in vitro growth competition assay for CRF07_BC was established. The in vitro growth competition assay was used to verify the relative fitness of the K103 N and K166R mutant strains to the wild strain. The relative fitness of BCEA2-K103 N/BCEA1 was 0.728282±0.16608, BCEA2-K166R/BCEA1 was 0.883385±0.19023, BCEA2-K103 N+ K166R/BCEA1 was 0.804604±0.06164. The relative fitness of K103 N resistant mutant strain and the wild strain was significantly different.Conclusions:An in vitro growth competition assay for the fitness of HIV-1 CRF07_BC virus was established. Based on the in vitro growth competition assay, the HIV-1 K103 N resistant mutant strain has reduced viral fitness.