Imprinted genes play a key role in regulating mammalian placental and embryonic development. Here, we generated glutaminyl-peptide cyclotransferase-knockout (Qpct^-/-) mice utilizing the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) platform and identified Qpct as a novel anti-angiogenic factor in regulating mouse placentation. Compared with Qpct^+/+ mice, placentae and embryos (Qpct^-/+ and Qpct^-/-) showed significant overgrowth at embryonic Day 12.5 (E12.5), E15.5, and E18.5. Using single-cell transcriptome analysis of 32 309 cells from Qpct^+/+ and Qpct^-/- mouse placentae, we identified 13 cell clusters via single-nucleus RNA sequencing (snRNA-seq) (8880 Qpct^+/+ and 13 577 Qpct^-/- cells) and 20 cell clusters via single-cell RNA sequencing (scRNA-seq) (6567 Qpct^+/+ and 3285 Qpct^-/- cells). Furthermore, we observed a global up-regulation of pro-angiogenic genes in the Qpct^-/- background. Immunohistochemistry assays revealed a notable increase in the number of blood vessels in the decidual and labyrinthine layers of E15.5 Qpct^-/+ and Qpct^-/- mice. Moreover, the elevation of multiple pairs of ligand-receptor interactions was observed in decidual cells, endothelial cells, and macrophages, promoting angiogenesis and inflammatory response. Our findings indicate that loss of maternal Qpct leads to altered phenotypic characteristics of placentae and embryos and promotes angiogenesis in murine placentae.
Animals ; Female ; Pregnancy ; Mice ; Placentation/genetics* ; Single-Cell Analysis ; Gene Expression Profiling ; Mice, Knockout ; Transcriptome ; Placenta/blood supply* ; Neovascularization, Pathologic/genetics* ; Genomic Imprinting ; Single-Cell Gene Expression Analysis ; Angiogenesis

ObjectiveTo investigate the effect of circSLC8A1_005 on the fibrotic phenotype of cardiac fibroblasts and the potential mechanism involved. MethodsThe effect of adenovirus-mediated overexpression of circSLC8A1_005 on the expression of fibrosis-related genes， collagen type I alpha 1 chain （Col1a1）， collagen type Ⅲ alpha 1 chain （Col3a1） and smooth muscle actin alpha 2 （Acta2）， in mouse cardiac fibroblasts （mCFs） were detected. The proliferation and migration activities of mCFs were detected by EdU and wound-healing assay， respectively. Dual luciferase reporter gene assay was performed to detect the activity of potential internal ribozyme entry site （IRES） in circSLC8A1_005. CircSLC8A1_005-translated protein， SLC8A1-605aa， and its intracellular distribution was identified by Western blot assay. The effect of SLC8A1-605aa protein on transcription activity of Sod2 gene was detected by the dual luciferase reporter gene assay. RNA binding protein immunoprecipitation （RIP） was utilized to verify the interaction between SLC8A1-605aa and superoxide dismutase 2 （Sod2） mRNA. Actinomycin D treatment was used to detect the effect of SLC8A1-605aa on Sod2 mRNA stability in mCFs. ResultsAn efficient adenovirus-mediated overexpression of circSLC8A1_005 was achieved in mCFs. The enforced expression of circSLC8A1_005 suppressed proliferation and migration of mCFs， and inhibited the expression of fibrosis-related genes in mCFs. The dual luciferase reporter gene assay revealed the activities of 2 IRES in circSLC8A1_005. Results of Western blot assay showed that circSLC8A1_005 could translate protein SLC8A1-605aa with the prospected molecular weight of 70 ku， which is predominantly distributed in the nucleus. Overexpression of the circSLC8A1_005 and SLC8A1-605aa could consistently inhibit the fibrotic phenotype of mCFs. SLC8A1-605aa could up-regulate superoxide dismutase 2 （Sod2） expression， but not at the transcriptional level. RIP assay indicated that SLC8A1-605aa could specifically interact with Sod2 mRNA， and the results of actinomycin D assay showed that SLC8A1-605aa could enhance the stability of Sod2 mRNA in mCFs. ConclusionCircSLC8A1_005 inhibits the fibrotic phenotype of cardiac fibroblasts via translating SLC8A1-605aa protein， and SLC8A1-605aa may be a potential target for the treatment of myocardial fibrosis.

AIM:To investigate the effect of circular RNA MYO9A-006(circMYO9A_006)on hypertrophic phenotype of cardiomyocytes and the underlying mechanism.METHODS:The effect of adenovirus-mediated overexpres-sion of circMYO9A_006 on the expression of hypertrophy-related proteins,including β-myosin heavy chain(β-MHC),skeletal muscle actin alpha 1(ACTA1)and atrial natriuretic peptide(ANP),was evaluated in neonatal mouse ventricular cardiomyocytes(NMVCs).Moreover,a neonatal rat ventricular cardiomyocyte(NRVC)model of phenylephrine(PE)-in-duced hypertrophy was established.The effect of circMYO9A_006 overexpression on NRVC size was ascertained using Phalloidin-iFluor 647 staining method.Dual-luciferase reporter assay was employed to measure the activity of potential in-ternal ribosome entry sites(IRES)in circMYO9A_006.The translation and intracellular location of the circMYO9A_006-translated protein,MYO9A-208aa,were verified using Western blot.To investigate the role of MYO9A-208aa in the ef-fect of circMYO9A_006 on the cardiomyocyte hypertrophic phenotype,we prepared and used the following adenoviruses:the recombinant circMYO9A_006-ORF adenovirus to express MYO9A-208aa,the recombinant circMYO9A_006-ATG-mut adenovirus that does not express MYO9A-208aa,the recombinant circMYO9A_006 adenovirus,and the adenovirus vector control.These were then employed to infect NRVCs.RESULTS:Successful adenovirus-mediated overexpression of circMYO9A_006 was observed in NMVCs.The increased expression of circMYO9A_006 notably reduced the expres-sion of hypertrophy-related proteins in NMVCs(P<0.01).Concurrently,overexpression of circMYO9A_006 substantially reduced the expression of hypertrophy-associated proteins and diminished the size of PE-induced NRVCs(P<0.05).Dual-luciferase reporter assay identified the activity of 2 IRES in circMYO9A_006.Western blot results indicated that circ-MYO9A_006 could produce the MYO9A-208aa protein with an anticipated molecular weight of 28 kD in NRVCs,primari-ly found in the cytoplasm.Elevated expression of both circMYO9A_006 and MYO9A-208aa consistently reduced the ex-pression of hypertrophy-associated proteins(P<0.01),and counteracted the enlarged size of PE-induced NRVCs(P<0.05).However,increased expression of circMYO9A_006-ATG-mut did not counteract the PE-induced hypertrophic phe-notype of NRVCs.CONCLUSION:circMYO9A_006 attenuates the hypertrophic phenotype of cardiomyocytes by synthe-sizing the MYO9A-208aa protein.

Molecularly defined renal carcinoma is a pathologic subtype of renal cell cancer (RCC) with a definite driver gene, which was first proposed in the 2022 WHO classification of tumors of the urinary system and male reproductive organs. The fumarate hydratase-deficient renal cell carcinoma and SMARCB1-deficient renal medullary carcinoma are highly aggressive and lethal subtypes. Due to the low incidence and lack of research on the mechanism, there is almost no effective treatment for these aggressive RCC subtypes. Cytoreductive nephrectomy or metastasectomy are important methods to improve the survival and quality of life of metastatic RCC patients under effective systemic therapy. However, for the highly aggressive RCC, the clinical value of the above surgical strategies is still unclear. In this review, we will discuss these problems in order to provide reference for the improvement of prognosis for patients with highly aggressive RCC subtypes.

Objective:To observe the inhibitory effect of lentivirus (LV)-mediated miR-191 on the proliferation and angiogenesis of human retinal vascular endothelial cells (hREC) cultured in vitro.Methods:The hREC cell lines were cultured in vitro and divided into control group, hypoxia group, LV-empty vector (LV-vector) group, and LV-miR-191 (LV-191) group. The LV-vector group and LV-191 group were transferred to the corresponding lentiviral vector respectively. Flow cytometry was used to detect cell transfection efficiency. Cell Counting Kit-8 (CCK-8) test was used to detect cell proliferation ability. Scarification test and invasion chamber (Transwell) test were used to detect cell migration ability. Matrigel test was used to detect cell lumen formation ability. Real-time quantitative polymerase chain reaction (qPCR) was used to detect the relative expression of miR-191 and relative mRNA expression of its downstream target genes p21, vascular endothelial growth factor (VEGF), cell division protein kinase (CDK) 6, cyclin-D1 (Cyclin D1). Independent sample t test was used for pairwise comparison. Results:The results of flow cytometry showed that the transfection efficiency of cells in the control group and the LV-191 group were 0.615% and 99.400%, respectively. The results of CCK-8, scarification, Transwell and Matrigel test showed that, compared with the control group, the number of cell proliferation ( t=6.130, 4.606), the cell mobility ( t=4.910, 6.702), the number of stained cells on the microporous membrane ( t=7.244, 6.724) and the lumen formation ability cells ( t=8.345, 9.859) were significantly increased in the hypoxia group and the LV-vector group ( P<0.01), while the LV-191 group showed completely opposite performance ( t=14.710, 6.245, 5.333, 5.892; P≤0.01). The qPCR test results showed that, compared with the control group and the LV-vector group, the relative expression of miR-191 mRNA in the cells of the LV-191 group was significantly up-regulated ( t=44.110, 42.680), the relative expression of Cyclin D1 mRNA ( t=29.940, 14.010) and CDK6 mRNA ( t=15.200, 7.645) decreased significantly, and the difference were statistically significant ( P<0.01); the relative expression of p21 mRNA increased, however, the difference was not statistically significant ( t=2.013, 2.755; P>0.05). There was no significant difference in the relative expression of VEGF mRNA in the 4 groups of cells ( F=0.966, P>0.05). Conclusions:LV-191 can inhibit the proliferation, migration and tubing of hREC by up-regulating p21 and down-regulating CDK6 and Cyclin D1.

Objective To preliminarily investigate the optimum scheme of thyroid CT flat scanning plan,to reduce the artifacts and the radiation dosage.Methods Patients who were involved in this research with thyroid CT flat scanning were divided into three groups:group A (conventional scan group),group B (dosage- reduced group)and group C (a combination of body posture change and dosage reduction).In group A (50 patients),the patients’bilateral arms were naturally drooped on both sides of the body during scanning (scan parameters:120 kV,250 mA).In group B (50 patients),the patients were scanned under the same circumstance with group A,but difference in scanning condition (scan parameters:100 kV,280 mA).In group C (50 patients),the patients kept one arm rise upward,another arm reach firmly toward the contralateral iliac bone during CT scan(scan parameters:100 kV,250 mA).CT attenuations,noise and SNR values around the soft tissue of thyroid gland,which was in the most severe thyroid artifacts (seventh cervical vertebrae to the first thoracic vertebrae level),were measured among the three groups.Meanwhile,the patient’s dose length product (DLP)and effective dosage (ED)were calculated.The images of the three groups were measured by double-blind method,i.e. severe artifacts,moderate artifacts,mild artifacts,and no artifacts.Results The image quality in group C was significantly higher than that of group A and group B (P＜0.05).There was no difference in subj ective quality between group A and group B (P＞0.05).The CT and noise values of group C were better than those of group A and group B (P＜0.05),while the radiation dosage was lower than that of group A and group B (P＜0.05).Conclusion When the thyroid CT scan is performed with the unilateral limb lifting and the other arm extending to the contralateral iliac bone,the thyroid band artifacts can be effectively reduced and decreasing the kV can reduce the radiation dosage.

Objective To observe the inhibitory effect of lentiviral vector miR-191 (LV-191) on retinal neovascularization (RNV) in mice model of oxygen-induced retinopathy (OIR).Methods Eighty healthy 7-day-old C57BL/6J mice were randomly divided into 5 groups including normal group, non-intervention group, normal saline (NS) group, LV-191 group and LV-green fluorescent protein (GFP) group, 16 mice in each group. The OIR model was established in the non-intervention group, NS group, LV-191 group and LV-GFP group. NS group, LV-191 group and LV-GFP group were given an intravitreal injection of 1 μl of NS, LV-191 and LV-GFP at the age of 12 days. No injection was performed in the non-intervention group. In normal group,newborn mouse were maintained in room air form P0 to P17, and no treatment was performed. Mice in all five groups were euthanized at P17. Retinal neovasculation (RNV) was evaluated by counting the number of pre-retinal neovascular cells and analysis of non-perfusion area area by immunofluorescent staining of the mouse retina. Real-time quantitative PCR (RT-PCR) to detect miR -191 and P21 expression of retinal tissue.Results In the LV-191 group, the non-perfusion area were both significantly smaller than those in non-intervention group, NS group and LV-GFP group (F=127.20,P<0.001). The number of pre-retinal neovascular cell nuclei in retinas from LV-191 group were obviously lower than those in the retinas from non-intervention group, NS group and LV-GFP group (F=31.71,P<0.05). RT-PCR showed that the LV-191 and P21 level of LV-191 group increased significantly than other groups (F=10.95, 15.60;P<0.05).Conclusion Intravitreal injection of LV-191 inhibits RNV in mice model of OIR possibly through up-regulating p21.

Objective To evaluate the effect of limb remote ischaemic preconditioning on pul-monary function in patients undergoing cardiac valve replacement surgery with cardiopulmonary by-pass.Methods Seventy patients,32 males and 38 females,aged 18-70 years,weighing 45-90 kg, ASA physical status Ⅱ or Ⅲ,scheduled for elective cardiac valve replacement surgery with cardiopul-monary bypass,were divided into 2 groups using a random number table,35 in each group.Patients in group R received three cycles of right upper-limb 5 min ischemia (blood-pressure cuff inflation to≥ 200 mm Hg)and 5 min reperfusion (blood-pressure cuff deflation to 0 mm Hg)at 10 min after in-tubation.In group C,the cuff was placed around the arm but not inflated.At 10 min after intubation (T0),at 1 h after aortic declamping (T1)and at 6 h (T2),12 h (T3),24 h (T4)after surgery,arte-rial blood was sampled to conduct gas analysis,PaO2/FiO2ratio and alveolar-arterial oxygen gradient (A-aDO2)were calculated,and the dynamic lung compliance (Cd)and static lung compliance (Cs) were also recorded.The occurrence of pulmonary adverse events was recorded until discharge. Results Compared with T0,PaO2/FiO2was decreased in the two groups at T1-T4,A-aDO2was de-creased at T2-T4,Cs and Cd were increased in group C at T3,and were increased in group R at T2, T3(P<0.05).Compared with group C,the Cs and Cd at T2,T3were increased in group R.There were no significant differences between the two groups in the PaO2/FiO2,A-aDO2at T0-T4.The oc-currence of the pulmonary adverse events was decreased significantly in group R than in group C (P<0.05).The occurrence of pulmonary adverse events was declined significantly in group R than in group C (P<0.05).Conclusion Limb remote ischemic preconditioning can improve the lung compli-ance and reduce the occurrence of the pulmonary adverse events in patients undergoing cardiac valve replacement surgery.

Objective: To investigate the application of Symptom Checklist-90 (SCL-90) in Chinese nurses. Methods: We performed a literature search for articles in Chinese on the mental health of nurses investigated using the SCL-90 that were published from January 2007 to December 2016. A total of 129 articles were included, involving 178 groups of valid samples as well as 36316 subjects in 27 provinces or cities in China. The metafor package of R 3.2.1 software and Excel were used for meta-analysis and data statistics. Results: Heterogeneity was identified for the nine factors included in the literature data (Q test: P<0.05; I² test >40%) , so a random effects model was used. For all factors except interpersonal relationship factor, the 95% confidence intervals of the pooled effect sizes did not include 0. For all factors except interpersonal relationship factor (Egger test: P<0.05) , no significant asymmetry or incompleteness appeared in the funnel plots. The fail-safe number of each factor was much larger than 5k+10. The merger scores and mean values of nine factors were higher than the national norm. Conclusion The SCL-90 is widely used in the study of mental status of Chinese nurses, but the SCL-90 norm for nurses is lacking.

Objective To retrospectively design an intensity?modulated radiotherapy ( IMRT) plan with split field and fixed jaw techniques for peripheral lung cancer with mediastinal lymph node metastasis, to compare dosimetric characteristics between the IMRT plans with fixed jaw and dynamic jaw, and to study lung protection by the plan with split field and fixed jaw. Methods Treatment plans were collected from 12 patients with peripheral lung cancer and mediastinal lymph node metastasis who were treated with IMRT in our hospital. All plans used the dynamic jaw technique. The plans with split field and fixed jaw were designed based on the identical computed tomography images and planning target volume ( PTV) . Each jaw position in split field depended on each separate PTV. The prescription dose was 60 Gy in 30 fractions. 95%PTV was planned to receive 100% of the prescription dose. Dosimetric parameters of PTV, conformity index ( CI) , heterogeneity index ( HI) , number of monitor units ( MUs) , and dose?volume values of the lung and heart were obtained from dose?volume histogram. Comparison between the two plans was made by paired t test. Results Both plans met clinical requirements. There were no significant differences in D2 , D98 , CI, or HI of PTV between the two plans ( all P>005) . Compared with the dynamic jaw plan, the fixed jaw plan had MUs increased by 15%?20%( P=0010) . The V5 , V10 , V20 , V30 , and mean dose for the whole lungs were significantly lower in the fixed jaw plan than in the dynamic jaw plan ( P=0000, 0000, 0000, 0002,0000) . The V5 , V20 , and mean dose for the healthy lung were also significantly lower in the fixed jaw plan than in the dynamic jaw plan ( P=0000,0017,0000) . There were no significant differences in dose?volume values for the spinal cord or heart between the two plans ( all P>005) . Conclusions IMRT with split field and fixed jaw is recommended for patients with peripheral lung cancer and mediastinal lymph node metastasis. The therapy to a certain extent reduces low?dose volume for the lung and the incidence of radiation?induced pneumonitis.