Objectives:To evaluate the imaging effects of the novel sympathetic nerve imaging agent 18F-FPMBBG in healthy volunteers and heart failure patients.Methods:Four healthy volunteers and four heart failure patients were selected to undergo 18F-FPMBBG positron emission tomography/computed tomography(PET/CT)dynamic imaging,the radioactivity distribution characteristics of 18F-FPMBBG in the heart and adjacent organs of the two groups were observed,and the uptake of 18F-FPMBBG by the left ventricular myocardium was compared in the two groups.Results:No adverse effects were observed in all subjects after intravenous injection of 18F-FPMBBG.In healthy volunteers,the heart uptake was rapid and stable,lung uptake was very low,and the blood pool and liver clearance were fast.The heart/liver uptake ratios at 30,60,and 90 minutes after injection were 2.33±0.81,3.29±0.90 and 3.80±1.07,respectively.The average standard uptake value(SUVmean)of 18F-FPMBBG in the heart failure group was significantly lower than that in the healthy volunteer group(P=0.003).The washout rate(WR)was significantly higher in the heart failure group([16.53±2.76]%vs.[3.88±4.51]%,P=0.003).Conclusions:18F-FPMBBG showed good imaging and diagnostic effects in the preliminary imaging of healthy subjects and heart failure patients,and it has the potential to become an ideal cardiac sympathetic nerve imaging agent.

This study investigated the specific immune response of BALB/c mice that was induced by a circular RNA (circRNA) vaccine expressing the herpes simplex virus type II (HSV-2) glycoprotein D (gD). The aim was to evaluate the immunological potential of this vaccine and lay a foundation for developing an mRNA vaccine against HSV-2. PCR and homologous recombination were employed to integrate the gD gene obtained from the pT7AMP-gD ectodomain plasmid into pUC57 to generate the recombinant plasmid pUC57-circ-gD, which was then sequenced and characterized. In vitro transcription and cyclization were performed on the template DNA to generate pUC57-circ-gD mRNA. To validate the formation of circular RNA, we cleaved the pUC57-circ-gD mRNA with RNase R and employed RT-PCR to validate the cyclization. The pUC57-circ-gD mRNA was then transfected into 293T cells. After 72 h, the cell supernatant was collected, and Western blotting was employed to measure the protein level of gD. Subsequently, the mRNA was encapsulated in lipid nanoparticles (LNPs) by microfluidic encapsulation. BALB/c mice were administrated with the encapsulated mRNA, and blood was collected from the fundus venous plexus after 21 and 35 days, and from the enucleated eyeballs after 49 days. Enzyme-linked immunosorbent assay was employed to measure the titers of antibodies, including virus-neutralizing antibodies. After 49 days, spleens were harvested and assessed for secretion of interferon-gamma (IFN-γ) by solid-phase enzyme-linked immunospot. The results showed successful construction and sequencing of the recombinant plasmid. RNase R digestion confirmed the presence of circular RNAs. Western blotting of the 293T cells transfected with the mRNA showed clear specific bands. The quality of the vaccine was tested by size exclusion chromatography-high performance liquid chromatography, which showed that the purity of the vaccine was about 90%. The mRNA-LNP showcased the particle size of 82.76 nm and an encapsulation rate of approximately 98%. Following three-dose vaccination, all immunized mice exhibited steady weight gain with 100% survival rate throughout the 28-day observation period, indicating no significant acute toxicity associated with the vaccine formulation. The immunized mice showed dose-dependent increases in serum IgG antibody titer and IFN-γ secretion by splenocytes and they were resistant to virus attacks. These findings indicate good immunogenicity and persistence of the pUC57-circ-gD mRNA vaccine, providing a reference for further studies on circRNA vaccines.
Animals ; Mice, Inbred BALB C ; RNA, Circular ; Mice ; Humans ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Antibodies, Viral/blood* ; HEK293 Cells ; Female ; Nanoparticles ; Plasmids

Objectives:To evaluate the imaging effects of the novel sympathetic nerve imaging agent 18F-FPMBBG in healthy volunteers and heart failure patients.Methods:Four healthy volunteers and four heart failure patients were selected to undergo 18F-FPMBBG positron emission tomography/computed tomography(PET/CT)dynamic imaging,the radioactivity distribution characteristics of 18F-FPMBBG in the heart and adjacent organs of the two groups were observed,and the uptake of 18F-FPMBBG by the left ventricular myocardium was compared in the two groups.Results:No adverse effects were observed in all subjects after intravenous injection of 18F-FPMBBG.In healthy volunteers,the heart uptake was rapid and stable,lung uptake was very low,and the blood pool and liver clearance were fast.The heart/liver uptake ratios at 30,60,and 90 minutes after injection were 2.33±0.81,3.29±0.90 and 3.80±1.07,respectively.The average standard uptake value(SUVmean)of 18F-FPMBBG in the heart failure group was significantly lower than that in the healthy volunteer group(P=0.003).The washout rate(WR)was significantly higher in the heart failure group([16.53±2.76]%vs.[3.88±4.51]%,P=0.003).Conclusions:18F-FPMBBG showed good imaging and diagnostic effects in the preliminary imaging of healthy subjects and heart failure patients,and it has the potential to become an ideal cardiac sympathetic nerve imaging agent.

Objective Changes in relevant indexes in the mouse model of early-onset ovarian insufficiency caused by Tripterygium wilfordii polyglycoside were analyzed,and the optimal time point for intervention was determined.Methods Forty female ICR mice were randomly divided into control and A,B,C,and D model groups with eight mice in each group.The control group was gavaged with purified water for 14 days(0.01 mL/10 g),and the remaining groups were administered a Tripterygium wilfordii polyglycoside suspension(80 mg/kg,0.01 mL/10 g)for 1 day(A model group),3 days(B model group),7 days(C model group),or 14 days(D model group),and samples were collected.Body weight and wet weights of the uterus and bilateral ovaries of mice were determined in each group.Serum FSH,LH,E2,P,AMH,INH-B,and T contents were measured using enzyme-linked immunoassays.HE staining was used to observe the number and developmental status of follicles and corpus luteum at all levels in mice of each group.TUNEL staining was used to detect the apoptosis in the ovaries of mice in each group.IHC detected expression of VEGFA,CD34,and EPO proteins in the ovaries of mice in each group.mRNA expression of HIF-1α,SDF-1,and CXCR4 in each group of mice was detected by PCR.Results Compared with the control group,changes in indicators in model A mice did not meet the POI modeling standard.The ovarian index,uterine index,and body weight of mice in the B model group were decreased significantly(P＜0.01),the weight of the C model group was decreased significantly(P＜0.01),and the ovarian index of the D model group was decreased significantly(P＜0.05).Serum contents of FSH and LH in B,C,and D model groups were increased(P＜0.05,P＜0.01),the E2,PROG,AMH,INH-B,and T contents were decreased(P＜0.01).The numbers of basal follicles,pre-sinus follicles,sinusoidal follicles,antral follicles,preovulatory follicles,and corpus luteum were decreased significantly(P＜0.05,P＜0.01)and the number of atresia follicles was increased significantly(P＜0.01)in B,C,and D model groups.The apoptotic area of TUNEL staining in A,B,C,and D model groups was increased significantly(P＜0.05,P＜0.01).Expression of CD34,VEGFA,and EPO in B,C,and D model groups was decreased significantly(P＜0.05,P＜0.01).mRNA expression of HIF-1α,SDF-1,and CXCR4 in A and B model groups was significantly increased(P＜0.05,P＜0.01).Compared with the B model group,the relevant indexes of C and D model groups were changed significantly,indicating that C and D models were more serious and tended to develop POF.Conclusions The B model group is the turning point of ovarian function from impaired POI to irreversible POF,suggesting that 3 days of administrating Tripterygium wilfordii polyglycoside is optimal to induce a POI disease model for effective drug intervention.

Hypoxia inducible factor-1α (HIF-1α), as a hypoxia inducible factor, affects women's reproductive function by regulating the development and excretion of follicles. HIF-1α induces glycolysis and autophagy in the granule cells by promoting oocyte development, regulating the secretion of related angiogenic factors, and improving follicle maturity. In addition, HIF-1α promotes the process of luteinization of follicular vesicles, maintains luteal function, and finally completes physiological luteal atrophy through cumulative oxidative stress. Dysfunction of HIF-1α will cause a series of pathological consequences, such as angiogenesis defect, energy metabolism abnormality, excessive oxidative stress and dysregulated autophagy and apoptosis, resulting in ovulation problem and infertility. This article summarizes the previous studies on the regulation of follicle development and excretion and maintenance of luteal function and structural atrophy by HIF-1α. We also describe the effective intervention mechanism of related drugs or bioactive ingredients on follicular dysplasia and ovulation disorders through HIF-1α, in order to provide a systematic and in-depth insights for solving ovulation disorder infertility.
Female ; Humans ; Atrophy/metabolism* ; Hypoxia ; Hypoxia-Inducible Factor 1, alpha Subunit/metabolism* ; Infertility/metabolism* ; Ovarian Follicle ; Ovulation

Objective:To observe the effect of acupuncture in the treatment of accommodative myopia in children.Methods:A total of 76 children with accommodative myopia who met the inclusion criteria were divided into a control group or a test group according to the random number table method,with 38 cases in each group.The control group was given education on eye hygiene,and the test group was treated with acupuncture twice a week for 2 months in addition to the intervention used in the control group.The patient's uncorrected visual acuity(UCVA),refraction,and axial length(AL)were measured before treatment and 1 month and 2 months after treatment.Results:After 1 month of treatment,there was no significant difference in the UCVA between the two groups(P＞0.05);after 2 months of treatment,the UCVA of the test group was better than that of the control group(P＜0.05).After 1 and 2 months of treatment,the refraction of the two groups was significantly different from that before treatment(P＜0.01),but there was no significant difference between the two groups(P＞0.05).After 1 and 2 months of treatment,the AL in the control group was increased compared with that before treatment(P＜0.05),while there was no significant change in the test group(P＞0.05),and there was no significant difference between the two groups(P＞0.05).Conclusion:Acupuncture treatment can improve UCVA in children with accommodative myopia.

Objective:To evaluate the effects of propofol on inflammatory responses in substantia nigra in mice with Parkinson′s disease (PD) and its relationship with α-synuclein (α-syn) expression.Methods:Thirty-three SPF healthy male C57BL/6 mice, aged 12 weeks, weighing 24-26 g, were divided into 3 groups ( n=11 each) using a random number table method: control group (group Con), group PD and propofol group (group Pro). In PD and Pro groups, 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) 30 mg/kg was intraperitoneally injected once a day for 5 consecutive days to induce PD.Propofol 50 mg/kg was intraperitoneally injected at 2 h after the last injection of MPTP in group Pro, while the equal volume of normal saline was given instead in Con and PD groups.The rotarod test was performed at 24 h after administration.The animals were then sacrificed and substantia nigra was removed for determination of contents of interleukin-1β (IL-1β) and tumor necrosis factor (TNF)-α (by enzyme-linked immunosorbent assay), the expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) and p-caspase-1 (by Western blot) and the expression of α-syn (by immunofluorescence staining). Results:Compared with Con group, the first fall-off time was significantly shortened, the number of falling off was increased, the contents of IL-1β and TNF-α were increased, and the expression NLRP3, p-caspase-1and α-syn was up-regulated in substantia nigra in group PD ( P<0.05). Compared with PD group, the first fall-off time was significantly prolonged, the number of falling off was decreased, the contents of IL-1β and TNF-α were decreased, and the expression NLRP3, p-caspase-1and α-syn was down-regulated in substantia nigra in group Pro ( P<0.05). Conclusion:Propofol can improve behaviors of the mice through inhibiting inflammatory responses in substantia nigra, and the mechanism is related to down-regulating the expression of α-syn.

The quality of public cardiopulmonary resuscitation training plays an important role in improving the survival rate of patients with cardiac arrest. Various forms of training have been carried out all over China, which plays a great role in promoting the work of cardiopulmonary resuscitation. However, there is still a lot of room for improvement in the quality management and effect sustainability of training. This paper reviews the current situation and deficiencies of quality management of public cardiopulmonary resuscitation training, and the role of training quality in enhancing people's self-confidence in learning and rescue, training contents, training methods, quality evaluation indicators, evaluation methods, and quality influencing factors and retraining time requirements, and so on. And it puts forward some practical suggestions on the quality management of public cardiopulmonary resuscitation training in China. Such as it will more emphasize standardized training, deliberate practice, proficient training, National Training, long-term maintenance of knowledge and skills, and using useful tools to improve the quality of cardiopulmonary resuscitation training, etc. In order to improve the training quality management level of the public, so that the trainees can really master cardiopulmonary resuscitation skills, so as to improve the rescue rate and survival rate of patients with cardiac arrest. To promote the sustainable development of people's health.

Objective:To evaluate the influence of the sleeve lengths and implant lengths on accuracy of static computer-assisted implant surgery (sCAIS).Methods:Twenty-eight models of bilateral mandibular single tooth loss were included. Fifty-five implants were placed under the guidance of sCAIS (Straumann Bone Level 4.1 mm×10 mm). According to the height of metal sleeve of static guide plate, 55 implants were divided into 11 groups (free hand group, 1 mm group, 2 mm group, 3 mm group, 4 mm group, 5 mm group, 6 mm group, 7 mm group, 8 mm group, 9 mm group, 10 mm group), with 5 implants in each group. Eight research models were included. Group with 5 mm sleeve guides were used to place implants of different length, (Straumann Bone Level width 4.1 mm, height was 8 mm, 10 mm and 14 mm), 5 implants in each group. Eighteen patients with mandibular single tooth loss were included in the Department of Oral Implantology, Tianjin Stomatological Hospital from October 2018 to June 2019. There were 10 males and 8 females, 18-46(33.7±7.9) years old. A total of 18 implants were implanted and divided into 3 groups (free hand group, 3 mm group and 5 mm group) with 6 implants in each group. Digital software was used to compare the implant positions before and after implantation. Non-parametric Kruskal-Wallis test or one-way ANOVA were used to analyze the results.Results:There was no significant difference in implant vertical deviation between different sleeve height groups (1-10 mm) and free hand group, but the neck deviation in free hand group[(1.04±0.13) mm] was significantly higher than that in different sleeve height groups (1-10 mm) ( P<0.05). The tip deviations of free hand group, 1 mm group and 2 mm group [(1.32±0.43), (0.83±0.10) and (0.78±0.11) mm, respectively] was significantly higher than that of 10 mm group [(0.31±0.14) mm]( P<0.05). The angle deviation of free hand group and 1 mm group (3.99°±0.85° and 2.59°±0.69°), respectively] was significantly higher than that of 10 mm group (0.61°±0.03°) ( P<0.05). The tip deviations of implants in the 14 mm group [(0.83±0.22) mm] was significantly higher than that in the 8 mm and 10 mm groups [(0.44±0.07) and (0.49±0.06) mm, respectively]. Clinical studies showed that there was no significant difference in neck deviation, tip deviation and angle deviation between 3 mm group and 5 mm group ( P>0.05), but deviations were significantly lower than those in free hand group ( P<0.05). Conclusions:The length of the sleeves has significant influence on the accuracy of the surgical guide. There was no significant difference in accuracy of the implant guide with 3 mm or 5 mm metal sleeves. The vitro study has some limitations and needs further systematic research.

G protein-coupled receptors (GPCRs) are membrane receptors whose agonist-induced dynamic conformational changes trigger heterotrimeric G protein activation, followed by GRK-mediated phosphorylation and arrestin-mediated desensitization. Cytosolic regions of GPCRs have been studied extensively because they are direct contact sites with G proteins, GRKs, and arrestins. Among various cytosolic regions, the role of helix 8 is least understood, although a few studies have suggested that it is involved in G protein activation, receptor localization, and/or internalization. In the present study, we investigated the role of helix 8 in dopamine receptor signaling focusing on dopamine D1 receptor (D1R) and dopamine D2 receptor (D2R). D1R couples exclusively to Gs, whereas D2R couples exclusively to Gi. Bioinformatic analysis implied that the sequences of helix 8 may affect GPCR-G protein coupling selectivity; therefore, we evaluated if swapping helix 8 between D1R and D2R changed G protein selectivity. Our results suggest that helix 8 is not involved in D1R-Gs or D2R-Gi coupling selectivity. Instead, we observed that D1R with D2R helix 8 or D1R with an increased number of hydrophobic residues in helix 8 relative to wild-type showed diminished β-arrestin-mediated desensitization, resulting in increased Gs signaling.
Arrestin ; Arrestins ; Computational Biology ; Cytosol ; Dopamine ; Family Characteristics ; GTP-Binding Proteins ; Membranes ; Phosphorylation ; Receptors, Dopamine D1 ; Receptors, Dopamine D2 ; Receptors, Dopamine