Chronic remote ischemic conditioning treatment in patients with chronic stable angina (EARLY-MYO-CSA): a randomized, controlled proof-of-concept trial.

BACKGROUND: Chronic remote ischemic conditioning (CRIC) has been shown to improve myocardial ischemia in experimental animal studies; however, its effectiveness in patients with chronic stable angina (CSA) has not been investigated. We conducted a proof-of-concept study to investigate the efficacy and safety of a six-month CRIC treatment in patients with CSA. METHODS: The EARLY-MYO-CSA trial was a prospective, randomized, controlled trial evaluating the CRIC treatment in patients with CSA with persistent angina pectoris despite receiving ≥ 3-month guideline-recommended optimal medical therapy. The CRIC and control groups received CRIC (at 200 mmHg) or sham CRIC (at 60 mmHg) intervention for 6 months, respectively. The primary endpoint was the 6-month change of myocardial flow reserve (MFR) on single-photon emission computed tomography. The secondary endpoints were changes in rest and stress myocardial blood flow (MBF), angina severity according to the Canadian Cardiovascular Society (CCS) classification, the Seattle Angina Questionnaire (SAQ), and a 6-min walk test (6-MWT). RESULTS: Among 220 randomized CSA patients, 208 (105 in the CRIC group, and 103 in the control group) completed the treatment and endpoint assessments. The mean change in MFR was significantly greater in the CRIC group than in the control group (0.27 ± 0.38 vs. - 0.04 ± 0.25; P < 0.001). MFR increased from 1.33 ± 0.48 at baseline to 1.61 ± 0.53 (P < 0.001) in the CRIC group; however, a similar increase was not seen in the control group (1.35 ± 0.45 at baseline and 1.31 ± 0.44 at follow-up, P = 0.757). CRIC treatment, when compared with controls, demonstrated improvements in angina symptoms assessed by CCS classification (60.0% vs. 14.6%, P < 0.001), all SAQ dimensions scores (P < 0.001), and 6-MWT distances (440 [400-523] vs. 420 [330-475] m, P = 0.016). The incidence of major adverse cardiovascular events was similar between the groups. CONCLUSIONS: CSA patients benefit from 6-month CRIC treatment with improvements in MFR, angina symptoms, and exercise performance. This treatment is well-tolerated and can be recommended for symptom relief in this clinical population. TRIAL REGISTRATION: [chictr.org.cn], identifier [ChiCTR2000038649].

Tspan5 promotes the EMT process to regulate the syncytialization of trophoblast cells by activating Notch signalling.

Placental trophoblastic cells play important roles in placental development and fetal health. However, the mechanism of trophoblastic cell fusion is still not entirely clear. The level of Tspan5 in the embryo culture medium was detected using enzyme-linked immunosorbent assay (ELISA). Fusion of BeWo cells was observed by immunofluorescence. Cell fusion-related factors and EMT-related factors were identified by qRT-PCR and western blotting. Notch protein repressor DAPT was used to verify the role of Tspan5 in BeWo cells. The expression of Tspan5 was significantly increased in embryo culture medium. The fusion of BeWo cells was observed after treatment with forskolin (FSK). Cell fusion-related factors (i.e. ß-hCG and syncytin 1/2) and Tspan5 were significantly increased after FSK treatment. In addition, FSK treatment promoted EMT-related protein expression in BeWo cells. Knockdown of Tspan5 inhibited cell fusion and EMT-related protein levels. Notch-1 and Jagged-1 protein levels were significantly upregulated, and the EMT process was activated by overexpression of Tspan5 in FSK-treated BeWo cells. Interestingly, blocking the Notch pathway by the repressor DAPT had the opposite results. These results indicated that Tspan5 could promote the EMT process by activating the Notch pathway, thereby causing cell fusion. These findings contribute to a better understanding of trophoblast cell syncytialization and embryonic development. Tspan5 may be used as a therapeutic target for normal placental development.

Pluronic F127 coating performance on PLGA nanoparticles: Enhanced flocculation and instability.

Nanoparticles (NPs) can be incorporated into hydrogels to obtain multifunctional hybrid systems to meet the delivery needs of different drugs. However, the stability of NPs in hydrogels is rarely revealed. In this article, we tried to explore the underlying mechanism of an interesting phenomenon that poly(lactic-co-glycolic acid) (PLGA) nanoparticles (PNPs) could flocculate and deposit in Pluronic F127 (F127) hydrogels at 4 °C. The results showed that this flocculation was relevant to the type of emulsifier formulated in PNPs, the particle materials and the F127 concentration, but independent of PLGA polymer end groups. Exactly, PNPs containing polyvinyl alcohol (PVA) as the emulsifier flocculated in F127 solution with a concentration above 15 %. The flocculated PNPs possessed increased particle size, decreased zeta potential, reduced hydrophobicity and an obvious coating layer, and these characteristics could be restored almost to the original state after two washes of flocculated PNPs with water. Moreover, the flocculation had no impact on the long-term size stability and drug-loading capacity of PNPs, and F127-treated PNPs showed improved cellular uptake than untreated PNPs. These results provide the evidence that adsorption of high concentrations of F127 on the surface of PNPs/PVA may lead to flocculation, and the flocculation is reversible by simply washing the flocs with water. To the best of our knowledge, this is the first study to scientifically explore the stability of PNPs in F127 hydrogels, providing theoretical and experimental support for the rational design and further development of nanoparticle-hydrogel composite.

The "L-Sandwich" Strategy for True Coronary Bifurcation Lesions: A Randomized Clinical Trial.

Background: This study explored the efficacy of the "L-sandwich" strategy, which involves the implantation of stents in the main vessel (MV) and shaft of the side branch (SB) with a drug-coated balloon (DCB) applied to the SB ostium, for coronary true bifurcation lesions. Methods and Results: Of 99 patients with true bifurcation lesions, 38 patients underwent the "L-sandwich" strategy (group A), 32 patients underwent a two-stent strategy (group B), and 29 patients underwent a single-stent + DCB strategy (group C). Angiography outcomes (late lumen loss [LLL], minimum lumen diameter [MLD]), and clinical outcomes (major adverse cardiac events [MACEs]) were analyzed. At 6 months, the MLD of the SB ostium in groups A and B were similar (P > 0.05) and group A larger than group C (P < 0.05). The LLL of group B was the largest among the three groups (P < 0.05). The MLD of the SB shaft in groups A and B were larger than in group C (P < 0.05). The LLL of the SB shaft in group C was the lowest (P < 0.05). Two patients in group B received target vessel revascularization at the 6-month followup (P > 0.05), and patients in the other groups had no MACEs. Conclusions: The "L-sandwich" strategy was feasible for the treatment of true coronary bifurcation lesions. It is a simpler procedure with similar acute lumen gain than the two-stent strategy, results in a larger SB lumen than the single-stent + DCB strategy, and it can also be used as a remedy for dissection following the single-stent + DCB strategy.

Targeted DNA demethylation of the ZNF334 promoter inhibits colorectal cancer growth.

Colorectal cancer (CRC) is a leading cause of cancer deaths worldwide. Aberrant regulation of DNA methylation in promoters of tumor suppressor genes or proto-oncogenes is one of the fundamental processes driving the initiation and progression of CRC. Zinc-finger proteins (ZNFs) are one of the most abundant groups of proteins and function in many important biological processes related to tumorigenesis. Herein, we detected abnormal hypermethylation of the ZNF334 gene in CRC tissues compared with normal tissues, and this modification downregulated the expression of ZNF334. Furthermore, ten-eleven translocation 1 (TET1) was identified to be involved in regulating the methylation level of ZNF334. Next, a dCas9-multiGCN4/scFv-TET1CD-sgZNF334-targeted demethylation system was constructed to reverse the expression of ZNF334 through sgRNA targeting the ZNF334 promoter. Both in vitro and in vivo experiments demonstrated the targeted demethylation system upregulated ZNF334 expression and inhibited CRC growth. Collectively, targeted DNA demethylation of the ZNF334 promoter sheds light on the precise treatment of CRC.

Risk Factors of Preterm Birth and Low Birth Weight in Singletons Conceived Through Frozen Embryo Transfer: A Retrospective Study.

Background: The risks of adverse perinatal outcomes in offspring conceived following frozen-thawed embryo transfer (FET) assisted reproductive technology (ART) are inconsistent. The aim of this study was to analyze the risk factors for preterm birth and low birth weight in singletons after FET. Methods: 386 FET cycles was conducted at the Reproductive Medicine Center of Meizhou People's Hospital. The relationship between clinical characteristics and outcomes (term birth and preterm birth, normal birth weight and low birth weight) was analyzed. Results: The rate of primary infertility, basal FSH and T levels, gestational age, birth weight, and proportion of male fetuses were significantly different in the preterm and full-term groups. Logistic regression analysis showed that high maternal age (≥35 years) (OR 3.652, 95% CI: 1.683-7.925, P=0.001), primary infertility (OR 2.869, 95% CI: 1.461-5.632, P=0.002), low FSH level (<6.215 mIU/mL) (OR 3.272, 95% CI: 1.743-6.144, P<0.001), and hormone replacement therapy (HRT) method (OR 2.780, 95% CI: 1.088-7.100, P=0.033) may increase risk of preterm birth after FET. Gestational age and birth weight were significantly different in fetuses with low birth weight (<2500g, n=38) and normal birth weight (≥2500g and <4000g, n=333). Logistic regression analysis showed that low basal FSH level (<6.215 mIU/mL) (OR 0.425, 95% CI: 0.209-0.865, P=0.018), and HRT method of endometrial preparation for FET (OR 0.272, 95% CI: 0.079-0.934, P=0.039) may reduce the risk of low birth weight after FET. Conclusion: High maternal age, primary infertility, low FSH level, HRT method of endometrial preparation for FET, and male fetus may increase risk of preterm birth after FET. In addition, primary infertility, low basal FSH level, and HRT method of endometrial preparation may reduce the risk of low birth weight after FET.

Risk Factors of Pregnancy Failure in Infertile Patients Undergoing Assisted Reproductive Technology.

Background: Infertile couples need to use assisted reproductive technology (ART) to give birth. However, pregnancy failure after ART is not uncommon. At present, the results of studies on the causes of pregnancy failure after ART are inconsistent. Methods: A retrospective cohort study involving 715 embryo transfer cycles was conducted at the Reproductive Medicine Center of Meizhou People's Hospital, from December 2015 to June 2022. According to the pregnancy, they were divided into clinical pregnancy group and pregnancy failure group. The relationship between demographic characteristics and pregnancy status between the two groups was analyzed. Results: The pregnancy failure rate after ART was 49.7% (355/715). There were statistically significant distribution differences of maternal age, paternal age, COH protocols, and number of embryos transferred between clinical pregnancy and pregnancy failure groups (all P<0.01). Multiple logistic regression analysis shows that high maternal age (>35 years old vs ≤35 years old: OR 2.173, 95% CI: 1.386-3.407, P=0.001), and GnRH-a short protocol (GnRH-a short protocol vs GnRH-a long protocol: OR 2.139, 95% CI: 1.127-4.058, P=0.020) may increase risk of pregnancy failure in ART pregnancies, while two embryos transferred (two embryos transferred vs one embryo transferred: OR 0.563, 95% CI: 0.377-0.839, P=0.005) may reduce risk of pregnancy failure. In addition, high maternal age, GnRH antagonist protocol, and GnRH-a short protocol may increase risk of implantation failure, while two embryos transferred may reduce risk of implantation failure. And high maternal age may increase risk of biochemical pregnancy. Conclusion: The risk of pregnancy failure increased in ART cycles with maternal age >35 years old and GnRH-a short protocol, while reduced with two embryos transferred.

Evaluation of an Injectable Hydrogel Based on Hyaluronic Acid-Chitosan/ß-Glycerophosphate-Loaded Mesenchymal Stem Cells in Enhancing the Therapeutic Efficacy of Myocardial Infarction.

Myocardial infarction (MI), which is due to cardiac dysfunction, results in morbidity and mortality. Moreover, the cellular activity of transplanted mesenchymal stem cells (MSCs) generally limits their therapeutic efficacy in the treatment of MI. Here, inject able hyaluronic acid-chitosan/ß-glycerophosphate (HA-CS/ß-GP) hydrogel-loaded MSCs are prepared, after which their effects on the treatment of MI are investigated. The synthesized HA-CS/ß-GP hydrogels exhibit swelling ratio, an in vitro degradation value, and a gelatin time of 82.19 ± 4.1, 88.18% ± 2.4%, and 9 s, respectively. Further, rheological studies revealed that the elastic modulus of the HA-CS/ß-GP hydrogels is ≥230 Pa, exhibiting large elastic to viscous modulus ratio, which indicates their mechanical strength. Furthermore, the in vitro 3T3 cell and MSC culture studies confirm the good biocompatibility of the HA-CS and HA-CS/ß-GP hydrogels. The implantation of the synthesized hydrogels in the mouse MI model considerably improves the therapeutic effect of the MSCs (enhanced cardiac function, reduced cardiomyocyte apoptosis, and increased vascularization) for the first time. The innovative synergistic strategy of combining injectable HA-CS and HA-CS/ß-GP hydro gels with MSCs may be suitable for the effective treatment of cardiac morbidity due to MIs.

The increased level of Tspan5 in villi suggests more proliferation and invasiveness of trophoblasts in tubal pregnancy.

OBJECTIVE: This study was to determinate the expression of Tspan5 in tubal ectopic implantation sites and to explore the correlation of the expressive level of Tspan5 at maternal-fetal interface and the occurrence of tubal ectopic pregnancy. STUDY DESIGN: This is a retrospective study. Trophoblastic and endometrial tissues were collected from tubal ectopic pregnancy(Total of 40), and intrauterine pregnancy(Total of 41), who had voluntary abortion, non-pregnancy women(Total of 12), who recieved an diagnostic uterine curettage before IVF-ET for male infertility. All samples were collected from women aged 23-40 years, from February 2012 to January 2014. Results 1. In human villi Tspan5 was primarily located in cytoplasm and on the surfaces of cytotroblasts(CTs) and extravillous trophoblast(EVCTs). The intensity of Tspan5 in tubal pregnancy was significantly higher than that in normal intrauterine pregnancy, showing significant differences (Mean of IOD:109.39 ±â€¯61.84 Vs. 89.04 ±â€¯36.44;t = 2.33, P = 0.023). 2. In human deciduas of intrauterine pregnancy or endometrium of tubal pregnancy and non-pregnancy Tspan5 expressed in cytoplasm and membrane of glandular epithelial cells. The expressive level of this protein was increased in tubal pregnancy than that in intrauterine pregnancy and non-pregnancy(Mean of IOD:144.18 ±â€¯106.22 Vs. 93.43 ±â€¯67.10, P = 0.037; 144.18 ±â€¯106.22 Vs. 88.56 ±â€¯33.24, P = 0.018). CONCLUSION: Our study indicated that the trophoblasts in tubal pregnancy showed more proliferative and invasive characteristics. Dysregulation of Tspan5 in decidual microenvironment may relate to the retention of embryo in fallopian tube. SUPPORT: This study was Supported by Science and Technology Planning Project of Guangdong Province.

Identification of a peripheral blood long non-coding RNA (Upperhand) as a potential diagnostic marker of coronary artery disease.

BACKGROUND: Long non-coding RNAs (lncRNAs) have been confirmed to be involved in the pathologi-cal processes of multiple diseases. However, the characteristic expression of lncRNAs in peripheral blood of coronary artery disease (CAD) patients and whether some of these lncRNAs can be used as diagnostic biomarkers for CAD requires further investigation. METHODS: Six healthy and CAD individuals were selected for microarray analysis, and 5 differentially expressed lncRNAs were selected and confirmed in the second cohort consisting of 30 control individu-als and 30 CAD patients with different SYNTAX scores. Upperhand were verified in the third cohort consisting of 115 controls and 137 CAD patients. RESULTS: Thirty one lncRNAs were differentially expressed between the two groups, among whom, 25 were upregulated in the CAD group and 6 were downregulated. Four of the selected five lncRNAs were significantly upregulated in the CAD group, and Upperhand had the largest area under the curve (AUC). The diagnostic value of Upperhand was tested further, and it remained having a high diagnostic value. CONCLUSIONS: The expression level of Upperhand in peripheral blood of CAD patients is significantly higher than in control individuals, and is correlated with severity of CAD. Upperhand is a potential diagnostic biomarker of CAD, and when combined with TCONS_00029157, diagnostic value slightly increased.

Hsa-circRNA11783-2 in peripheral blood is correlated with coronary artery disease and type 2 diabetes mellitus.

OBJECTIVE: The purpose of this study was to identify the expression characteristics of circular RNAs in the peripheral blood of coronary artery disease patients and type 2 diabetes mellitus patients. METHODS: Circular RNA in the peripheral blood from 6 control individuals, 6 coronary artery disease patients, 6 type 2 diabetes mellitus patients and 6 coronary artery disease combined with type 2 diabetes mellitus patients was collected for microarray analysis, and a further independent cohort consisting of 20 normal individuals, 20 type 2 diabetes mellitus subjects and 20 coronary artery disease subjects was used to verify the expression of five circular RNAs chosen for further analysis. The findings were then tested in a third cohort using quantitative real-time polymerase chain reaction. RESULTS: In total, 40 circular RNAs differentially expressed between the three experimental groups and the control group were identified by microarray analysis: 13 were upregulated in the experimental groups, while 27 were downregulated. Of the five circular RNAs chosen for further analysis, three were significantly downregulated in the experimental groups. The crude odds ratios and adjusted odds ratios of hsa-circRNA11783-2 showed significant differences in both the coronary artery disease group and type 2 diabetes mellitus group. We then verified hsa-circRNA11783-2 in the third cohort, and it remained closely related to both coronary artery disease and type 2 diabetes mellitus. CONCLUSION: Hsa-circRNA11783-2 is closely related to both coronary artery disease and type 2 diabetes mellitus.

The Diagnostic Value of Whole Blood lncRNA ENST00000550337.1 for Pre-Diabetes and Type 2 Diabetes Mellitus.

This study aims to investigate long noncoding RNA (lncRNA) as biomarker for pre-diabetes and T2DM. LncRNAs in the peripheral blood of 6 healthy individuals and 6 T2DM patients were collected for microarray analysis. Then 5 candidate biomarkers from the differentially expressed lncRNAs were chosen and verified in a larger independent cohort (control group=20; pre-diabetes group=20; and T2DM group=20). The diagnostic capacity of ENST00000550337.1 was further tested in the third cohort (control group, n=60; pre-diabetes group, n=63; and T2DM group, n=64). A total of 17 lncRNAs were found to be differentially expressed between the 2 groups. 14 lncRNAs of these were upregulated in T2DM patients and 3 were downregulated. 5 upregulated lncRNAs were selected as potential biomarkers and verified in the second cohort, and the expression levels of 3 lncRNAs increased gradually from the control group to the pre-diabetes group to the T2DM group. The diagnostic value of ENST00000550337.1 was then tested in the third cohort, and its high diagnostic value for pre-diabetes and T2DM was confirmed. LncRNA ENST00000550337.1 is a potential diagnostic biomarker for pre-diabetes and T2DM.