[摘 要] 目的：探讨乙醛脱氢酶1A1（ALDH1A1）mRNA负载树突状细胞（DC）疫苗体外诱导抗鼻咽癌干细胞免疫反应的能力。方法：体外转录ALDH1A1 mRNA并电转染至人外周血DC，构建ALDH1A1-DC疫苗，转染EGFP mRNA为对照。流式细胞术检测DC表型、淋巴细胞亚群、细胞毒性T 淋巴细胞（CTL）增殖和细胞因子分泌。分选ALDH+和ALDH- C666-1细胞，检测干细胞标志物（CD24、CD133）和干性基因（OCT4、SOX2、Nanog）的表达水平，采用CCK-8法评估ALDH1A1-DC疫苗诱导的特异性免疫杀伤能力。结果：EGFP-DC荧光表达率显著高于未转染DC（P < 0.000 1），ALDH1A1-DC抗原提呈分子HLA-ABC、共刺激分子CD86、CD40和CCR7表达与未转染DC相比显著增强（P = 0.001 3、P = 0.005 9、P = 0.000 4及P = 0.001 9）。与单独淋巴细胞组（T）及空载DC诱导的淋巴细胞组（DC-T）相比，ALDH1A1-DC诱导的CTL比例最高（均P < 0.000 1），分裂指数最高（P = 0.000 2、P = 0.000 3），且CD8+CD69+（P = 0.000 5、P < 0.000 1）和CD8+ IFN-γ+（P = 0.012 6、P = 0.001 9）细胞亚群比例均显著提高。ALDH1A1-DC-T细胞对ALDH+C666-1悬浮细胞球的杀伤效果显著优于ALDH-细胞球（P = 0.001 0），且与其他各组淋巴细胞相比表现出较强的抗ALDH+C666-1悬浮细胞球的免疫效应（P = 0.001 0，P < 0.000 1）。然而，HLA-ABC阻断后其对ALDH+C666-1细胞的特异性杀伤效应显著降低（P < 0.000 1）。结论：mRNA转染DC后可翻译蛋白并促进DC成熟，增强其诱导特异性CTL免疫反应的能力。

Objective To investigate the effects of electroacupuncture for the restoration of muscle regeneration and the secretion of exosomes around acupoints in a model of erector spinae muscle injury.Methods Forty SPF-grade male SD rats were randomly divided into blank group,model group,electroacupuncture group,and electroacupuncture+exosome inhibitor group,with 10 rats per group.Except for the blank group,the erector spinae muscle injury models were established in other groups by intramuscular injection of 0.5%bupivacaine.The blank control group received no treatment,whereas the rats in the electroacupuncture and electroacupuncture+exosome inhibitor groups were treated with electroacupuncture at"Weizhong"(BL40)and"Shenshu"(BL23)acupoints,respectively,stimulation was applied daily for 7 consecutive days,with each session lasting 20 minutes.The parameters used were a sparse-dense wave waveform,a frequency of 2/10 Hz,and a current intensity of 1 mA.The exosome inhibitor GW4869(3 g/L,50 μL per acupoint)was injected 1 h before each electroacupuncture in the electroacupuncture+exosome inhibitor group.After intervention,the erector spinae muscles were collected and observed by HE and Masson staining for morphological changes.The expression of paired box gene 7(Pax7)and recombinant myogenic differentiation(MyoD)was detected by immunohistochemistry,while the expression of myogenin(MyoG)and myosin heavy chain(MyHC)proteins was detected by western blotting.The serum exosomes of rats in each group were extracted and identified by transmission electron microscopy and nanoparticle tracking analysis,and the expression of Alix,differentiation cluster 63(CD63),and tumor susceptibility gene 101(TSG101)proteins were detected by Western blotting.Results Compared with the blank group,the model group,the electroacupuncture group,and the electroacupuncture+exosome inhibitor group exhibited spinae muscle fiber fragmentation,degeneration,necrosis,and inflammatory cell infiltration in HE staining.The result of Masson staining showed that collagen fiber hyperplasia was increased.The model group showed increased expression of MyoD,Pax7,MyoG,MyHC,and CD63,while TSG101 expression was downregulated(P<0.05).In the electroacupuncture group,the expression of MyoD,Pax7,Alix,and TSG101 was elevated(P<0.05),and the expression of MyHC and CD63 was decreased(P<0.05).The electroacupuncture+exosome inhibitor group displayed increased expression of MyHC,Alix,and TSG101(P<0.05),and the expression of CD63 was decreased(P<0.05).Compared with the model group,the electroacupuncture group and the electroacupuncture+exosome inhibitor group showed reduced muscle fiber degeneration,necrotic areas,and inflammatory cell infiltration as observed in HE staining,along with decreased collagen fiber hyperplasia in Masson staining.Specifically,the electroacupuncture group demonstrated increased expression of MyoD,Pax7,MyoG,Alix,and TSG101(P<0.05),and the expression of CD63 was decreased(P<0.05).The electroacupuncture+exosome inhibitor group displayed downregulated expression of Pax7,MyoG,MyHC,and CD63(P<0.05),and the expression of Alix and TSG101 was regulated(P<0.05).Compared with the electroacupuncture+exosome inhibitor group,the electroacupuncture group exhibited less muscle fiber degeneration and necrosis,reduced inflammatory cell infiltration in HE staining,and decreased stained collagen fibers in Masson staining.The electroacupuncture group showed increased expression of MyoD,Pax7,MyoG,MyHC,Alix,and CD63(P<0.05).Conclusion Electroacupuncture can up-regulate the expression of Pax7 and MyoD,and promote the regeneration of erector spinae muscles,which may be related to stimulating the secretion of exosomes around the acupoint.Exosomes may be an important mediator for the efficacy of acupuncture.

Objective:To analyze the incidence rate and disability-adjusted life year (DALY) rate of Chlamydia trachomatis ( C. trachomatis) infection in women aged 15-49 years in China from 1990 to 2021, identify the vulnerable populations, and provide evidence for the prevention and control of C. trachomatis infection. Methods:The data from Global Burden of Disease Study 2021 were used to analyze the incidence rate and DALY rate of C. trachomatis infection in women aged 15-49 years in China from 1990 to 2021 by using Joinpoint regression model. The age-period-cohort analysis method was used to decompose the age effect, period effect and cohort effect of incidence rate and DALY rate of C. trachomatis infection. Results:From 1990 to 2021, the incidence rate of C. trachomatis infection increased from 5 892.02/100 000 in 1990 to 7 402.23/100 000 in 2021 in women aged 15-49 years in China. The DALY rate increased from 1.92/100 000 person-years in 1990 to 2.00/100 000 person-years in 2021. Joinpoint regression showed that, from 1990 to 2021, the annual incidence of C. trachomatis infection in Chinese women age aged 15-49 years increased by 0.74% [average annual percent change (AAPC) =0.74%, 95% CI: 0.67%-0.81%, t=21.94, P<0.001] and the annual average DALY rate increased by 0.11% (AAPC=0.11%, 95% CI: -0.09%-0.30%, t=1.06, P=0.287). Age-period-cohort model analyses showed that women aged 30-34 years were at high risk to be infected with C. trachomatis and women aged 40-44 years were at high risk to have high DALY. The period effect showed that the risk for C. trachomatis infection began to increase in 1990, peaked during 2000-2004, then gradually decreased. During 2015-2019, the infection risk increased again, while the risk for high DALY showed a decreasing trend. The cohort effect showed that the risks for C. trachomatis infection and DALY were lower in later birth cohorts. Conclusions:From 1990 to 2021, the incidence rate and DALY rate of C. trachomatis infection showed fluctuations in women aged 15-49 years in China. It is necessary to pay close attention to the prevention and control of C. trachomatis infection in women aged 30-34 years.

Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tarii(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.

Objective:To analyze the incidence rate and disability-adjusted life year (DALY) rate of Chlamydia trachomatis ( C. trachomatis) infection in women aged 15-49 years in China from 1990 to 2021, identify the vulnerable populations, and provide evidence for the prevention and control of C. trachomatis infection. Methods:The data from Global Burden of Disease Study 2021 were used to analyze the incidence rate and DALY rate of C. trachomatis infection in women aged 15-49 years in China from 1990 to 2021 by using Joinpoint regression model. The age-period-cohort analysis method was used to decompose the age effect, period effect and cohort effect of incidence rate and DALY rate of C. trachomatis infection. Results:From 1990 to 2021, the incidence rate of C. trachomatis infection increased from 5 892.02/100 000 in 1990 to 7 402.23/100 000 in 2021 in women aged 15-49 years in China. The DALY rate increased from 1.92/100 000 person-years in 1990 to 2.00/100 000 person-years in 2021. Joinpoint regression showed that, from 1990 to 2021, the annual incidence of C. trachomatis infection in Chinese women age aged 15-49 years increased by 0.74% [average annual percent change (AAPC) =0.74%, 95% CI: 0.67%-0.81%, t=21.94, P<0.001] and the annual average DALY rate increased by 0.11% (AAPC=0.11%, 95% CI: -0.09%-0.30%, t=1.06, P=0.287). Age-period-cohort model analyses showed that women aged 30-34 years were at high risk to be infected with C. trachomatis and women aged 40-44 years were at high risk to have high DALY. The period effect showed that the risk for C. trachomatis infection began to increase in 1990, peaked during 2000-2004, then gradually decreased. During 2015-2019, the infection risk increased again, while the risk for high DALY showed a decreasing trend. The cohort effect showed that the risks for C. trachomatis infection and DALY were lower in later birth cohorts. Conclusions:From 1990 to 2021, the incidence rate and DALY rate of C. trachomatis infection showed fluctuations in women aged 15-49 years in China. It is necessary to pay close attention to the prevention and control of C. trachomatis infection in women aged 30-34 years.

Objective: To investigate the effects of C1q tumor necrosis factor⁃related protein 3 (CTRP3) ⁃enhanced Sendai virus (SeV) vector⁃overexpressing Gata4 , Mef2c , and Tbx5 (SeVGMT) in the treatment of myocardial in⁃farction (MI) mice and to analyze whether ubiquitin carboxyl⁃terminal hydrolase L1 (UCHL1) mediates this thera⁃peutic pathway. Methods: The mice were divided into 7 groups ( n = 12) : Sham group , MI group , SeVGMT group , CTRP3⁃Lv group , UCHL1 ⁃sh group , SeVGMT + CTRP3⁃Lv group , and SeVGMT + CTRP3⁃Lv + UCHL1 ⁃sh group. In the Sham group , only the skin was incised without ligation , while the coronary artery was ligated 2 - 3 mm below the left atrial appendage in mice in other groups. PBS was injected at three points in the myocardial infarction boundary in the Sham and MI groups 30 minutes after ligation. Mice in other groups were injected with SeVGMT , CTRP3-Lv , or UCHL1-sh according to their grouping. Four weeks after treatment , fractional shortening (FS) , ejection fraction (EF) , left ventricular end-diastolic diameter (LVIDd) , left ventricular end-systolic diameter (LVIDs) , heart rate (HR) , mean arterial pressure (MAP) , serum creatine kinase isoenzyme MB (CK-MB) , myocardial troponin I ( cTnI) and lactate dehydrogenase ( LDH) levels , and myocardial tumor necrosis factor-α (TNF-α ) , interleukin-1β (IL-1β) and interleukin-6 (IL-6) levels in mice were detected. The pathological changes of myocardial tissue were detected by 2 , 3 , 5-triphenyltetrazolium chloride ( TTC) , hematoxylin-eosin ( HE) , Masson trichrome and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. The mRNA expressions of CTRP3 and UCHL1 were detected by qRT-PCR. The protein expressions of CTRP3 , UCHL1 , collagen Ⅰ , collagen Ⅲ , Bcl-2-associated X (Bax) and B-cell lymphoma/leukemia-2 (Bcl-2) in myocardial tissue were detected by Western blot or immunohistochemical staining. Results: Compared with SeVGMT group and CTRP3-Lv group , the levels of EF , FS , HR and MAP in SeVGMT + CTRP3-Lv group increased (P < 0. 05) . The levels of LVIDd , LVIDs , CK-MB , cTnI , LDH , TNF-α , IL-1β and IL-6 decreased (P < 0. 05) . MI area , fibrosis area and TUNEL positive rate decreased (P < 0. 05) , the protein levels collagen Ⅰ , collagen Ⅲ and Bax decreased (P < 0. 05) , and Bcl-2 protein levels increased (P < 0. 05) . The mRNA and protein levels and relative staining intensity of CTRP3 and UCHL1 increased (P < 0. 05) . Compared with SeVGMT + CTRP3-Lv group , the addition of UCHL1-sh treatment ( SeVGMT + CTRP3-Lv + UCHL1-sh group) significantly weakened the influence of SeVGMT + CTRP3-Lv on the above indexes (P < 0. 05) . Conclusion CTRP3 mediated UCHL1 enhances the therapeutic effect of SeVGMT reprogrammed CFs on MI in mice.

Objective To systematically evaluate the risk factors for new-onset atrial fibrillation (NOAF) after off-pump coronary bypass grafting (OPCABG). Methods PubMed, EMbase, Web of Science, The Cochrane Library, Wanfang data, CBM, VIP, and CNKI databases were systematically searched by computer to collect studies related to the risk factors for NOAF after OPCABG from the establishment of the database to July 2023. Literature screening and quality evaluation were conducted independently by two researchers. The Newcastle-Ottawa Scale (NOS) was used to evaluate the quality of the literature. RevMan 5.3 and Stata15.0 were used for meta-analysis. Results Finally, 19 case-control studies related to the risk factors for NOAF after OPCABG were included, all of which were high-quality literature with NOS score≥6 points, with a total of 7019 subjects. The results of meta-analysis showed that the following factors were associated with NOAF after OPCABG: (1) the patient’s own factors: age (MD=3.51, 95%CI 2.39 to 4.63, P<0.01); (2) preoperative factors: history of hypertension (OR=1.17, 95%CI 1.04 to 1.32, P=0.01), history of myocardial infarction (OR=1.21, 95%CI 1.06 to 1.38, P<0.01), history of percutaneous coronary intervention (OR=2.22, 95%CI 1.03 to 4.77, P=0.04), EuroSCOREⅡ score (MD=0.59, 95%CI 0.25 to 0.94, P<0.01), low-density lipoprotein (MD=0.11, 95%CI 0.02 to 0.20, P=0.02), left atrial diameter (MD=1.64, 95%CI 0.24 to 3.04, P=0.02); (3) postoperative and treatment factors: left ventricular end-diastolic diameter (MD=1.16, 95%CI 0.33 to 1.99, P<0.01), left ventricular ejection fraction (MD=0.90, 95%CI 0.07 to 1.73, P=0.03), mechanical ventilation time (MD=2.78, 95%CI 1.65 to 3.90, P<0.01), B-type natriuretic peptide (MD=219.67, 95%CI 27.46 to 411.88, P=0.03), ICU retention time (MD=7.07, 95%CI 5.64 to 8.50, P<0.01). Conclusion The existing evidence shows that age, history of hypertension, history of myocardial infarction, history of percutaneous coronary intervention, preoperative EuroSCOREⅡscore, preoperative low-density lipoprotein, preoperative left atrial diameter, postoperative left ventricular end-diastolic diameter, postoperative left ventricular ejection fraction, postoperative mechanical ventilation time, postoperative B-type natriuretic peptide, and postoperative ICU retention time are risk factors for NOAF after OPCABG. Clinical attention should be paid to the above factors to achieve early identification, thereby reducing the incidence of NOAF after OPCABG and improving the clinical prognosis of patients.

Objective To investigate the efficiency and mechanism of C1q tumor necrosis factor-related protein 3(CTRP3)on reprogramming of cardiac fibroblasts(CFs)into induced cardiomyocyte-like cells(iCMs)by Sendai virus(SeV)vector overexpressing Gata4,Mef2c and Tbx5(SeVGMT).Methods CFs were divided into Control,NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh groups.NC-Lv,CTRP3-Lv,NC-sh,and CTRP3-sh were transfected into CFs using Lipofectamine 3000 reagent for 48 hours.Lipofectamine 3000 reagent was then mixed with SeVGMT and incubated at room temperature for 48 hours,the culture medium was then replaced,and cells were cultured for 21 days.Cell morphology was observed under a microscope at 0,3,7,14,and 21 days.Expression levels of the myocardial-specific proteins α-myosin heavy chain(α-MHC),α-actin,cardiac troponin T(cTnT),connexin 43(Cx43),cardiac muscle α-actin(Actc1),and myosin heavy chain 6(Myh6)were detected at different time points by immunofluorescence,quantitative reverse transcription-polymerase chain reaction,and Western blot,and the proportions of beating cells at different time points were calculated.Results The relative fluorescence intensity and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs in each group increased with increasing culture time(P＜0.05),with significantly higher expression levels of myocardial-specific proteins at 14 days of culture than at 7 days(P＜0.05).The relative fluorescence intensities and mRNA and protein levels of α-MHC,α-actin,cTnT,Cx43,Actc1,and Myh6 in CFs at 3,7,14,and 21 days of culture were significantly increased in the CTRP3-Lv group compared with the NC-Lv group(P＜0.05),but were significantly decreased in CFs in the CTRP3-sh group compared with the NC-sh group(P＜0.05).Beating cells appeared in CFs in each group at 7 days of culture.The proportion of beating cells in each group increased with increasing culture time(P＜0.05),and the proportion was significantly higher at 14 days than at 7 days(P＜0.05).The proportion of beating cells among CFs was increased in the CTRP3-Lv group at 7,14,and 21 days of culture compared with the NC-Lv group(P＜0.05),while the proportion of beating cells in the CTRP3-sh group was decreased compared with the NC-sh group(P＜0.05).Conclusions CTRP3 can enhance SeVGMT reprogramming of CFs into iCMs.

Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tarii(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.