ObjectiveTo characterize the systemically absorbed components of Zhengqi prescription （ZQP） and investigate its therapeutic effects and mechanisms against cancer-related fatigue （CRF） through an integrated approach of network pharmacology and animal experiments. MethodsThe systemically absorbed components of ZQP were identified by ultra-performance liquid chromatography coupled with quadrupole-Orbitrap mass spectrometry （UPLC-Q-Exactive Orbitrap MS）. Network pharmacology was employed to predict the potential targets and signaling pathways involved in the anti-CRF effects of ZQP. A mouse model of paclitaxel-induced CRF， mimicking breast cancer-related fatigue， was established. Mice were administered different doses of ZQP extract. The therapeutic efficacy was evaluated based on fatigue-related indicators， behavioral test results， and energy metabolism parameters. Western blot analysis was conducted to validate the regulation of core signaling pathways. ResultsA total of 48 systemically absorbed components were identified. Network pharmacology analysis revealed 124 intersecting targets （including Akt1） and implicated signaling pathways such as phosphatidylinositol 3-kinase/protein kinase （PI3K/Akt）. Pharmacodynamic studies with the established CRF model demonstrated that ZQP alleviated fatigue symptoms. Specifically， ZQP-treated mice exhibited increased exhaustive swimming time （P<0.05）， prolonged struggling time in the tail suspension test， and enhanced movement duration and distance in the open field test （P<0.05， P<0.01）. Furthermore， ZQP treatment decreased serum lactate and blood urea nitrogen levels （P<0.05， P<0.01）， increased liver and muscle glycogen content （P<0.05， P<0.01）， and upregulated the expression of phosphorylated （p）-Akt and p-PI3K in the gastrocnemius muscle （P<0.05， P<0.01）. ConclusionZQP exerts significant therapeutic effects on paclitaxel-induced breast cancer-related fatigue by regulating the PI3K/Akt signaling pathway. Furthermore， this study provides a preliminary foundation for subsequent quality control of the formula and further investigation into its mechanism against CRF.

OBJECTIVE: To observe the clinical efficacy of electroacupuncture (EA) at "four points of sacral region" for mild-to-moderate benign prostatic hyperplasia (BPH) with lower urinary tract symptoms (LUTS). METHODS: A total of 58 patients with BPH/LUTS were randomly divided into a sacral four-point EA group (29 cases, 1 case dropped out) and a conventional EA group (29 cases, 1 case dropped out). EA was applied at bilateral points 0.5 cun lateral to the sacrococcygeal joint and Huiyang (BL35) in the sacral four-point EA group; and was applied at Guanyuan (CV4), Zhongji (CV3), Qugu (CV2) and bilateral Shuidao (ST28), Sanyinjiao (SP6), Zusanli (ST36) in the conventional EA group. Both groups received continuous wave, 2 Hz in frequency, 30 min a time, once every other day, 3 times a week for 4 weeks. Before treatment, after 2 and 4 weeks of treatment, and in follow-up of 1 month after treatment completion, the international prostate symptom score (IPSS), the overactive bladder symptom score (OABSS), the quality of life (QOL) score were observed; before and after treatment, the prostate volume (PV) was measured by abdominal ultrasound; and the clinical efficacy was evaluated after treatment in the two groups. RESULTS: Compared before treatment, the scores of IPSS, OABSS and QOL were decreased after 4 weeks of treatment and in follow-up in both groups (P<0.01, P<0.05). In the sacral four-point EA group, the scores of IPSS and QOL after 4 weeks of treatment and in follow-up were lower than those in the conventional EA group (P<0.05, P<0.01), and the OABSS score in follow-up was lower than that in the conventional EA group (P<0.05). After 4 weeks of treatment and in follow-up, the reductions of IPSS and OABSS scores compared before treatment in the sacral four-point EA group were larger than those in the conventional EA group (P<0.01, P<0.05); After 2, 4 weeks of treatment and in follow-up, the reductions of QOL score compared before treatment in the sacral four-point EA group were larger than those in the conventional EA group (P<0.01). No significant difference in PV was observed after treatment between the two groups (P>0.05). The total effective rate was 75.0% (21/28) in the sacral four-point EA group, which was higher than 39.3% (11/28) in the conventional EA group (P<0.01). CONCLUSION EA at "four points of sacral region" can effectively improve the LUTS, overactive bladder symptom and quality of life in patients with mild-to-moderate BPH, and has good short-term and long-term efficacy, although it can not reduce prostate volume.
Humans ; Male ; Electroacupuncture ; Prostatic Hyperplasia/complications* ; Aged ; Middle Aged ; Lower Urinary Tract Symptoms/physiopathology* ; Acupuncture Points ; Treatment Outcome ; Quality of Life

Objective: To explore the correlation between allogeneic red blood cell (RBC) transfusion and healthcare-associated infections (HAIs) in patients undergoing coronary artery bypass grafting (CABG) during the perioperative period. Methods: A single-center retrospective cohort of 1,170 patients undergoing isolated CABG was analyzed. Multivariable logistic regression and restricted cubic splines (RCS) were employed to explore the nonlinear association between perioperative RBC transfusion (from intraoperative period to 72 hours postoperatively) and HAIs. Results: Among the 1,170 CABG patients, 109 patients (9.2%) received RBC transfusion during the operation or within 3 days after the operation. The risk of HAIs in those who received ≥4 units of RBCs during and within 3 days after the operation was 6.89 times higher than that in the non-transfusion group (95% CI: 3.65-17.20). Furthermore, there was a nonlinear threshold effect between the blood transfusion volume and postoperative HAIs (inflection point: 7.8 units). When the transfusion volume was ≤7.8 units, the risk of HAIs increased by 61% for each additional unit transfused (OR=1.61, 95% CI: 1.21-2.15). Beyond this threshold, no statistically significant association was observed (P=0.289). Conclusion: Perioperative RBC transfusion in CABG patients is associated with an increased incidence of HAIs. The perioperative blood transfusion volume has a curvilinear relationship with the risk of postoperative HAIs. When the blood transfusion volume is ≤7.8 units, the blood transfusion volume has a dose-dependent relationship with postoperative infection, with higher blood transfusion volumes correlating with greater postoperative infection risk. When the blood transfusion volume is >7.8 units, the relationship between the two is not statistically significant. The preventive effect of reducing RBC transfusion on HAIs requires further validation in the future.

OBJECTIVE To study the effects of Ro 64-6198,a selective nociceptin/orphanin FQ receptor(NOPR)agonist,on heroin self-administration and drug-seeking behavior in rats.METHODS Rats were trained to self-administer heroin intravenously at a dose of 0.05 mg·kg-1 under a fixed ratio 1(FR1)reinforcement schedule.Heroin motivation was assessed using a progressive ratio(PR)schedule.Firstly,a stable heroin self-administered rat model was established before the effects of Ro 64-6198 on heroin rewarding under the FR1 schedule were observed.After three days of self-administration recovery training,the effects of Ro 64-6198 on heroin reward motivation were observed under the PR3-4 schedule.Following extinction,the reinstatement of heroin seeking induced by either conditioned cues or heroin priming was evaluated in rats withdrawn from self-administration.The expressions of cAMP response element-binding protein(CREB)and brain-derived neurotrophic factor(BDNF)in the ventral tegmental area(VTA)were analyzed using Western blotting,while the expression of the NOPR in neurons in the VTA was examined through immunofluorescence staining.RESULTS Pretreatment with 3 mg·kg-1 Ro 64-6198 significantly reduced active responses and heroin infusions during FR1 testing,as well as decreased breakpoints,indicating reduced motivation under the PR schedule.At a dose of 1 mg·kg-1,Ro 64-6198 markedly attenuated the reinstatement of heroin-seeking behavior induced by conditioned cues or heroin priming.Furthermore,the administration of SB-612111,an NOPR antagonist,blocked the inhibitory effects of Ro 64-6198 on cue-induced heroin-seeking,although SB-612111 alone had no effect on heroin-seeking behavior.Ro 64-6198 treatment also suppressed the reduction of both phos-phorylated CREB(p-CREB)and BDNF levels in the VTA and the decreased expression of NOPR and p-CREB in dopaminergic neurons of the VTA.CONCLUSION These results demonstrate that Ro 64-6198 can mitigate heroin-seeking behavior through NOPR activation and CREB/BDNF pathway in the VTA.This study is expected to offer evidence for its potential as a clinical treatment for heroin addiction and relapse.

OBJECTIVE To study the effects of Ro 64-6198,a selective nociceptin/orphanin FQ receptor(NOPR)agonist,on heroin self-administration and drug-seeking behavior in rats.METHODS Rats were trained to self-administer heroin intravenously at a dose of 0.05 mg·kg-1 under a fixed ratio 1(FR1)reinforcement schedule.Heroin motivation was assessed using a progressive ratio(PR)schedule.Firstly,a stable heroin self-administered rat model was established before the effects of Ro 64-6198 on heroin rewarding under the FR1 schedule were observed.After three days of self-administration recovery training,the effects of Ro 64-6198 on heroin reward motivation were observed under the PR3-4 schedule.Following extinction,the reinstatement of heroin seeking induced by either conditioned cues or heroin priming was evaluated in rats withdrawn from self-administration.The expressions of cAMP response element-binding protein(CREB)and brain-derived neurotrophic factor(BDNF)in the ventral tegmental area(VTA)were analyzed using Western blotting,while the expression of the NOPR in neurons in the VTA was examined through immunofluorescence staining.RESULTS Pretreatment with 3 mg·kg-1 Ro 64-6198 significantly reduced active responses and heroin infusions during FR1 testing,as well as decreased breakpoints,indicating reduced motivation under the PR schedule.At a dose of 1 mg·kg-1,Ro 64-6198 markedly attenuated the reinstatement of heroin-seeking behavior induced by conditioned cues or heroin priming.Furthermore,the administration of SB-612111,an NOPR antagonist,blocked the inhibitory effects of Ro 64-6198 on cue-induced heroin-seeking,although SB-612111 alone had no effect on heroin-seeking behavior.Ro 64-6198 treatment also suppressed the reduction of both phos-phorylated CREB(p-CREB)and BDNF levels in the VTA and the decreased expression of NOPR and p-CREB in dopaminergic neurons of the VTA.CONCLUSION These results demonstrate that Ro 64-6198 can mitigate heroin-seeking behavior through NOPR activation and CREB/BDNF pathway in the VTA.This study is expected to offer evidence for its potential as a clinical treatment for heroin addiction and relapse.

OBJECTIVE To investigate the role of the complement C3/C3aR signaling pathway in the prefrontal cortex and colon neuroglia cell interactions during meth-amphetamine(METH)addiction,to observe the effects of TLR4 inhibitors as well as complement C3 elimination on METH reward and relapse behavior,and to explore the neuroinflammatory mechanisms of complement C3 acti-vation in METH addiction.METHODS ①A 14 d and 28 d rat METH addiction model was established to observe the effects of TLR4 antagonist ibudilast 3 mg·kg-1 and 10 mg·kg-1 on self-administration,reward motivation,relapse,and natural reward behavior in METH-trained 14 d rats and the effects of 0.02 mg·kg-1 complement C3 antago-nist on self-administration behavior in METH-trained 28 d rats.② Differences in the expression of TLR4,NF-κB,GRP94,C3,cathepsin L,CD68,and GFAP in the pre-frontal cortex of each group were examined using West-ern blotting.③ In addition,the expression of ATF6 in the prefrontal cortex of each group and the effects on neuro-nal and microglia/macrophage INOS,CD206 GRP94,and complement C3/C3aR.RESULTS ① Endoplasmic reticulum stress occurred in neurons and microglia after METH exposure depending on GRP94 and unfolded pro-tein responses to the ATF6 pathway.In addition,it acti-vates the TLR4-NF-κB pathway.② Microglia with high complement C3/C3aR expression in the prefrontal cortex were recruited to synaptic pruning and phagocytic responses around neurons with high GRP94,comple-ment C3/C3aR expression and these effects were blocked by complement C3 antagonists.③ In the rec-tum,GRP94 functions as a molecular chaperone for com-plement C3 and cathepsin L.Crosstalk occurs between enteric neurons high in GRP94,complement C3,and macrophages high in C3aR,located in the submucosa,lamina propria,and muscular,respectively,and all of these effects are blocked by complement C3 antago-nists.④ Treatment with the TLR4 antagonist ibudilast inhibits self-administration,reward motivation,and cue-or METH-priming in METH-trained 14 d rats,but fails to affect natural reward behavior.Ibudilast treatment attenu-ates the TLR4-NF-κB inflammatory pathway and comple-ments C3/C3aR pathway in the prefrontal cortex.CON-CLUSION Activation of the complement C3/C3aR signal-ing pathway by TLR4-NF-κB inflammatory signaling in the prefrontal cortex mediates the METH addiction pro-cess,providing an experimental basis for the clinical treatment of METH addiction,and targeting TLR4/NF-κB inflammatory signaling and complement C3/C3aR may be a new way to intervene in METH addiction.

Objective: To investigate association between adolescent depressive symptoms with circadian rhythm and emotion regulation strategies, and to provide the basis for mental health education for depression. Methods: CES-D, Morning and Evening Questionnaire-5 (MEQ-5) and Emotion Regulation scale (ERS) were administered to 2 398 students from 6 middle schools in Chengdu, Langzhong and Leshan of Sichuan Province. SPSS 21.0 was used to data processing and anlysis. Results: About 37.9% (909/2 398) of adolescents reported depressive symptoms. Prevalence of depressive symptom was higher in female students( χ 2= 25.15 , P <0.01), rural adolescents( χ 2=15.45, P <0.01), adolescents aged 15-18 compared to aged 12-14( χ 2=187.24, P < 0.01 ). There was significant difference in rate of depressive symptoms among adolescents with different circadian rhythms( χ 2= 55.19 , P < 0.01 ), with definite evening rhythm preference was the highest(57.1%). Prevalence of depressive symptoms significantly varied by sleep duration( χ 2=141.99, P <0.01), and were highest in adolescents with sleep duration <6 h(69.4%). The scores of suppression dimension in depressed adolescents were significantly higher than that of non depressive group, while the scores of reappraisal dimension were significantly higher in non depressive group than that of depressive group. Multivariate Logistic regression analysis showed that gender( OR =1.60), age( OR=2.29), suppression( OR = 1.13 ), sleep duration <6 h( OR =5.17), sleep duration 6-8 h ( OR =2.88) were positively associated with depressive symptoms in adolescents. Moderate type( OR =0.53), morning type ( OR =0.55) and cognitive reappraisal ( OR =0.90) were associated with lower rate of depressive symptoms( P <0.05). Conclusion Sleep rhythm delay, lack of sleep and emotion suppression in adolescents are associated with higher risk for depression. Regular sleep habits and reasonable emotion regulation might help to prevent adolescent depression.

Objective To observe the effects of adiponectin (APN) on anxiety-and depression-like behaviors occurred in APP/PS1/tau triple transgenic Alzheimer's disease (3xTg-AD) model mice and investigate the possible mechanism.Methods The 9-month-old 3xTg-AD mice and wild type (WT) mice were randomly divided into four groups (n=8 for each group):WT+Saline,WT+APN,3xTg-AD+Saline and 3xTg-AD+APN groups.The mouse was chronically intracerebroventricular injection of APN or saline under free moving condition.Elevated plus maze task was used to evaluate the anxiety-like behaviors.Sucrose preference test and tail suspension test were used to evaluate the depression-like behaviors.Western blot was used to detect the protein expression levels of p-AMPK,AMPK,p-GSK-3β (Ser9),GSK-3β and SIRT1 in the hippocampus of each mouse.Results In the elevated plus maze test,the time percentage spent in open arms of 3xTg-AD+Saline mice was ((4.2±2.7) ％),which significantly lower than that in WT+Saline group ((10.1±4.1)％) (q=4.349,P＜0.01) and in 3xTg-AD+APN group ((9.7±3.3)％) (q=4.065,P＜0.01).In the sucrose preference test,the sucrose preference percentage in 3xTg-AD+Saline group((48.8±5.3) ％) was lower than that in WT+Saline group ((60.9±6.7) ％) (q=4.522,P＜0.01) and in 3xTg-AD+APN group ((59.3±6.2)％) (q=3.911,P＜0.05).As for tail suspension test,the immobility time percentage in 3xTg-AD+Saline group((40.7±9.9)％) was higher than that in WT+Saline group ((27.1±4.8)％) (q=5.257,P＜0.001) and in 3xTg-AD+APN group ((31.4±6.0)％) (q=3.624,P＜0.05).The result of Western blot showed that there was no significant difference in expression level of AMPK and GSK-3β among four groups.However,compared with that in WT+Saline group,the expression levels of p-AMPK (q=3.586,P＜0.05),p-GSK-3β (Ser9) (q=3.125,P＜0.05),and SIRT1 (q=3.044,P＜0.05) in 3xTg-AD+Saline group were significantly decreased.In addition,compared with that in 3xTg-AD+Saline group,the expression levels of p-AMPK (q=3.374,P＜0.05) and p-GSK-3β (Ser9) (q=3.063,P＜0.05) in 3xTg-AD+APN group were obviously up-regulated without affecting the SIRT1.Conclusion Adiponectin can effectively alleviate the anxiety-and depression-like behaviors of 9-month-old 3xTg-AD mice by up-regulating the protein expression of p-AMPK and p-GSK-3β (Ser9) in the hippocampus.

The aim of the present study is to construct the recombinant eukaryotic expression and short hairpin RNA (shRNA) expression vectors of microRNA-21 and phosphatase and tensin homolog deleted on chromosome ten (PTEN). According to the gene sequence of microRNA-21 and PTEN, we designed and synthesized two pairs of single-stranded siRNA oligonucleotides and PCR primers. After annealing, the double-stranded DNA oligonucleotides were cloned into vector Psilencer4. 1-CMV. In addition, the gene sequences encoding pre-miR-21 and PTEN were amplified from colorectal cancer cell HCT-116 by RT-PCR. Then the PCR products were digested with restrictive endonuclease enzyme and cloned into vector pEGFP-N1. The constructed recombinant vectors were identified by restrictive digestion and DNA sequence analysis. The positive clone was confirmed by double enzyme digestion, and the enzyme fragments were consistent with the vector and purpose gene sequence. DNA sequencing confirmed that the purpose oligonucleotide fragments were correctly inserted in to the eukaryotic expression plasmids. It could be concluded that the microRNA-21 and PTEN eukaryotic expression and shRNA expression vectors have been successfully constructed, providing a foundation for further study on the effect of miR-21 on human colorectal cancer.
Base Sequence ; Cell Line, Tumor ; Cloning, Molecular ; Colonic Neoplasms ; genetics ; pathology ; Genetic Vectors ; genetics ; Humans ; MicroRNAs ; biosynthesis ; genetics ; Molecular Sequence Data ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; RNA, Small Interfering ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

We adjusted medical statistics teaching contents and teaching methods according to the characteristics of different students and based on the current status of domestic medical statics teaching.Implementation effect was summarized in order to provide references for the teaching reform of medical statistics.