Salusins: advance in cardiovascular disease research.

Salusins are discovered in 2003 and divided into salusin-α and salusin-ß, which are bioactive peptides with hemodynamic and mitotic activity and mainly distributed in plasma, urine, endocrine glands and kidneys. A large number of studies have shown that salusins can regulate lipid metabolism, inflammatory response and vascular proliferation. Despite the profound and diverse physiological properties of salusins, the exact mechanism of their cardiovascular effects remains to be determined. The potential mechanisms of action of salusins in cardiovascular-related diseases such as atherosclerosis, hypertension, heart failure, myocardial infarction and myocarditis, and their use as biomarkers of cardiovascular disease are discussed. This review aims to provide a new strategy for the diagnosis and prevention of clinical cardiovascular diseases.

Effect of Controlling Nutritional Status Score (CONUT) and Prognostic Nutritional Index (PNI) on patients after spinal tuberculosis surgery.

The controlling nutritional status (CONUT) score and prognostic nutrition index (PNI) are immune-nutritional biomarkers that are related to clinical prognosis. Previous studies have reported using them to predict the prognosis of traumatic brain injury, tumours and other diseases. The purpose of this study was to evaluate the relationship between the PNI and CONUT score and the one-year prognosis of patients with spinal tuberculosis (STB). In this study, the clinical characteristics of 97 patients with STB who underwent debridement and internal fixation at our institution between 2015 and 2020 were retrospectively analysed. According to the receiver operating characteristic (ROC) curve, patients were divided into two groups: a high CONUT group and a low CONUT group. Patients were also divided into a high PNI group and a low PNI group. One-year postoperative prognosis was evaluated by the clinical cure standard. Patients in the favourable group were younger and had a lower rate of pneumonia and urinary tract infection, higher PNI and lower CONUT score than those in the favourable group (P < 0.05). There was an obvious correlation between the PNI and CONUT score (r = - 0.884, P < 0.05). The areas under the curve (AUCs) of the CONUT score and PNI for predicting unfavourable prognosis were 0.888 (95% CI 0.808-0.943, P < 0.001) and 0.896 (95% CI 0.818-0.949, P < 0.001), respectively. The adjusted odds ratios (ORs) of the CONUT score and PNI for predicting unfavourable outcomes were 2.447 (95% CI 1.518-4.043, P < 0.001) and 0.689 (95% CI 0.563-0.843, P < 0.001), respectively. Higher CONUT scores and a lower PNI were associated with adverse outcomes in patients with spinal tuberculosis, and the CONUT score and PNI might be independent predictors of adverse outcomes of spinal tuberculosis postoperatively.

An orally available Mpro inhibitor is effective against wild-type SARS-CoV-2 and variants including Omicron.

Emerging SARS-CoV-2 variants continue to cause waves of new infections globally. Developing effective antivirals against SARS-CoV-2 and its variants is an urgent task. The main protease (Mpro) of SARS-CoV-2 is an attractive drug target because of its central role in viral replication and its conservation among variants. We herein report a series of potent α-ketoamide-containing Mpro inhibitors obtained using the Ugi four-component reaction. The prioritized compound, Y180, showed an IC50 of 8.1 nM against SARS-CoV-2 Mpro and had oral bioavailability of 92.9%, 31.9% and 85.7% in mice, rats and dogs, respectively. Y180 protected against wild-type SARS-CoV-2, B.1.1.7 (Alpha), B.1.617.1 (Kappa) and P.3 (Theta), with EC50 of 11.4, 20.3, 34.4 and 23.7 nM, respectively. Oral treatment with Y180 displayed a remarkable antiviral potency and substantially ameliorated the virus-induced tissue damage in both nasal turbinate and lung of B.1.1.7-infected K18-human ACE2 (K18-hACE2) transgenic mice. Therapeutic treatment with Y180 improved the survival of mice from 0 to 44.4% (P = 0.0086) upon B.1.617.1 infection in the lethal infection model. Importantly, Y180 was also highly effective against the B.1.1.529 (Omicron) variant both in vitro and in vivo. Overall, our study provides a promising lead compound for oral drug development against SARS-CoV-2.

Viral resistance to VRC01-like antibodies with mutations in loop D and V5 from an HIV-1 B' subtype infected individual with broadly neutralization activity.

Recently we identified the VRC01-like antibody DRVIA7(A7) from an HIV-1 B' subtype-infected individual (DRVI01) with broad neutralization activity, and almost all viruses from the individual were resistant to both VRC01 and A7 lineage antibodies. Here, we identified and characterized a panel of HIV-1 variants with resistance to VRC01 and A7 using site-directed mutagenesis and swapping amino acid fragments of gp120. Site-directed mutagenesis revealed that E279D/R282K/N460A/T464N of gp120 from DRVI01 produced VRC01-susceptible variants. Multiple mutations significantly increased the neutralization sensitivity to VRC01. Residues N464 located at the tip of the V5 loop were considered irrelevant to the neutralization of VRC01. For DRVI01-derived viruses, the single N464T change fully produced VRC01-resistant variants; conversely, a single T464N mutation generated VRC01-susceptible variants. Alanine scanning revealed that the N464 residue plays a vital role in binding with VRC01. Neutralizing assays against A7 lineage antibodies showed that DRVI01-derived viruses with multiple mutations could be neutralized by A7 lineage antibodies with different neutralizing breadths. Combining the changes in loops D and V5 produced variants that were totally sensitive variants to A7 lineage antibodies.

Potential Biomarkers in Early Pregnancy for Predicting Gestational Diabetes Mellitus and Adverse Pregnancy Outcomes.

BACKGROUND: Gestational diabetes mellitus (GDM) is typically diagnosed based on a 75-g oral glucose tolerance test conducted at 24 - 28 weeks of pregnancy. A method for earlier diagnosis is needed. The present study aimed to identify one or more blood biomarkers detected within the first trimester that can predict the occurrence of GDM and pregnancy outcome. METHODS: This retrospective study included 2,116 pregnant women who underwent examination and delivery in our hospital between January 2018 and December 2019. The predictive value of various clinical measurements in early pregnancy for predicting GDM and pregnancy outcome was analyzed. RESULTS: The fasting plasma glucose (FPG), vitamin A, vitamin E, glycosylated hemoglobin (HbA1c), total cholesterol (TC), triglyceride (TG), uric acid, free thyroxine (FT3), anti-peroxidase antibody (TPOAb), and ferritin levels differed significantly between the GDM and non-GDM groups (all p < 0.05). The area under the receiver operating characteristic curve for FPG in GDM diagnosis was 0.766 (95% confidence interval [CI] 0.717 - 0.814, p < 0.001). The odds ratios (ORs) for FPG and TG for GDM prediction were 1.318 (95% CI 1.228 - 1.416) and 2.050 (95% CI 1.203 - 3.493), respectively. The ORs for FPG, vitamin A, and vitamin E for pregnancy outcome prediction were 1.214 (95% CI 1.123 - 1.268), 0.717 (95% CI 0.601 - 0.886), and 0.852 (95% CI 0.761 - 0.954), respectively. CONCLUSIONS: Screening of blood biomarkers in early pregnancy may be useful for predicting, and thus preventing, GDM and adverse pregnancy outcomes. Immediate intervention is recommended if an elevated FPG (> 4.7 mmol/L) or TG (> 1.83 mmol/L) level is detected in early pregnancy, and vitamin A, vitamin E, and FT3 levels need to be maintained within normal ranges throughout pregnancy.

ACOX1, regulated by C/EBPα and miR-25-3p, promotes bovine preadipocyte adipogenesis.

Acyl-coenzyme A oxidase 1 (ACOX1) is the first and rate-limiting enzyme in peroxisomal fatty acid ß-oxidation of fatty acids. Previous studies have reported that ACOX1 was correlated with the meat quality of livestock, while the role of ACOX1 in intramuscular adipogenesis of beef cattle and its transcriptional and post-transcriptional regulatory mechanisms remain unclear. In the present study, gain-of-function and loss-of-function assays demonstrated that ACOX1 positively regulated the adipogenesis of bovine intramuscular preadipocytes. The C/EBPα-binding sites in the bovine ACOX1 promoter region at -1142 to -1129 bp, -831 to -826 bp, and -303 to -298 bp were identified by promoter deletion analysis and site-directed mutagenesis. Electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) further showed that these three regions are C/EBPα-binding sites, both in vitro and in vivo, indicating that C/EBPα directly interacts with the bovine ACOX1 promoter and inhibits its transcription. Furthermore, the results from bioinformatics analysis, dual luciferase assay, site-directed mutagenesis, qRT-PCR, and Western blotting demonstrated that miR-25-3p directly targeted the ACOX1 3'UTR (3'UTR). Taken together, our findings suggest that ACOX1, regulated by transcription factor C/EBPα and miR-25-3p, promotes adipogenesis of bovine intramuscular preadipocytes via regulating peroxisomal fatty acid ß-oxidation.

Artesunate inhibits atherosclerosis by upregulating vascular smooth muscle cells-derived LPL expression via the KLF2/NRF2/TCF7L2 pathway.

Lipoprotein lipase (LPL) plays a central role in hydrolyzing triglyceride and its deficiency leads to atherosclerosis. Artesunate (ART), a derivative of artemisinin, has been demonstrated that ART reduces the formation of atherosclerotic plaques. However, it remains unclear whether ART-alleviated atherosclerotic lesion is involved in regulating lipid metabolism. ApoE-/- mice were fed a high-fat diet to form atherosclerotic plaques and then injected with artesunate or not. Oil Red O, HE and Masson staining were performed to assess atherosclerotic plaques. Both Western blot and qRT-PCR were applied to detect protein expression. The Luciferase reporter gene and Chromatin immunoprecipitation assays were used to assess the interaction between proteins. Immunofluorescence assay was performed to show the localization of target proteins. In vitro, our data shown that ART increased LPL expression and inhibition of NRF2 blocked the binding of TCF7L2 to LPL promoter region in VSMCs. Downregulated Klf2 could decrease the nuclear enrichment of NRF2, TCF7L2 and LPL expression. In vivo, ART decreased atherosclerotic plaque formation and increased VSMC counts and LPL expression within atherosclerotic plaques. We observed the reduced tendency of serum lipids, and increased in serum LPL activity in mice. In support of vitro data, the markedly increased KLF2, TCF7L2 and LPL expression have been detected in aorta. Our study suggests that ART may be a novel therapeutic drug for inhibition of atherosclerotic plaque formation. The molecular mechanism may involve in upregulation of LPL expression via the KLF2/NRF2/TCF7L2 pathway in VSMCs.

Value of Procalcitonin in Neonatal Infection within 24 Hours after Birth: a Retrospective Cohort Study.

BACKGROUND: Neonatal infections, especially neonatal pneumonia, are clinically common and have a high mortality rate. Early diagnosis and the duration of appropriate antibiotic treatment are critical. PCT is an indication of infection and may be valuable. METHODS: This is a retrospective cohort of 269 neonates within 24 hours after birth, analyzing the value of procalcitonin, C-reactive protein, and white blood cell count in neonatal infections, especially neonatal pneumonia, and antibiotic therapy. RESULTS: The median of PCT, CRP, and WBC in the severely infected group, neonatal pneumonia group, neonatal infection group, and non-infectious disease group were (1.76, 5.25, 15.8), (0.20, 0.53, 13.8), (0.22, 3.64, 10.4), and (0.15, 0.39, 10.6), respectively. In ROC curves, PCT had an area under the curve (AUC) of 0.64 (95% CI, 0.49 - 0.0.79); CRP had an AUC of 0.61 (95% CI, 0.49 - 0.74); WBC had an AUC of 0.78 (95% CI, 0.67 - 0.88). There was a significant difference between the neonatal pneumonia with PCT results group and the neonatal pneumonia without PCT results group, p < 0.001. The median of antibiotic treatment was 4.0 d (95% CI 3.7 - 4.8) in the neonatal pneumonia with PCT results group vs. 4.9 d (95% CI 4.5 - 5.6) in the standard group; p < 0.001. CONCLUSIONS: PCT helps identify neonate infections and grades of infections and assists pediatricians in deciding when to stop antibiotic treatment; PCT and WBC help improve the accuracy of neonatal pneumonia diagnosis.

Amaryllidaceae alkaloids from Crinum latifolium with cytotoxic, antimicrobial, antioxidant, and anti-inflammatory activities.

Four novel and potently bioactive Amaryllidaceae alkaloids, 4,8-dimethoxy-cripowellin C (1), 4,8-dimethoxy-cripowellin D (2), 9-methoxy-cripowellin B (3), and 4-methoxy-8-hydroxy-cripowellin B (4), together with one known alkaloid, cripowellin C (5) were isolated from the 95% EtOH extract of the bulbs of Crinum latifolium. Structural elucidation of all the compounds were performed by spectral methods such as 1D and 2D (1H-1H COSY, HMQC, and HMBC) NMR as well as spectroscopy high resolution mass spectrometry. All isolates were in vitro evaluated for their cytotoxic activity against seven lung cancer cell lines, in addition to antimicrobial activity for eight bacteria, scavenging potential using ABTS·+ and DPPH test, and anti-inflammatory activity for Cox-1 and Cox-2 which had not previously been tested for crinane-type alkaloids with the cleavage between C-1 and C-13. Consequently, alkaloids 1-5 exhibited potent cytotoxicity against all of seven tested tumor cell lines with (IC50 < 30 nM). Alkaloids 3 and 4 displayed the significant antimicrobial activity with IC50 values <0.50 mM and antioxidant activity in the ABTS·+ and DPPH test. Additionally, Alkaloids 1-5 exhibited comparable inhibition of Cox-1 (>64%) and Cox-2 (>90%) with positive control.

[Construction of murine CXCR3 gene transfected cell line, identifiction and study of its biological function].

AIM: To construct recombinant murine CXCR3 gene retroviral vector and obtain L929-mCXCR3 gene transfected cell line for stably expressing murine CXCR3. We further study on L929-mCXCR3 migration effect resulted from interaction by CXCR3 and its ligand IP-10. METHODS: One female BALB/c mouse (7 weeks) was injected with 0.5 mg Con A intravenously (i.v.) via a tail vein. Twelve hours later the mouse was sacrificed and the spleen was removed.The spleen was pressed through a 150 microm stainless steel mesh. The isolated splenocytes were cultured in RPMI1640 supplemented with 50 U/mL human IL-2 for 3 days.Total RNA was extracted with TRIzol. Murine CXCR3 gene of full length was amplified by RT-PCR, then, it was inserted into retrovirus vector pEGZ-term. The recombinant vector together with its helper virus vector were co-transfected into package cell 293T with Lipofectamine(TM); 2000.The supernatant of 293T was collected for infecting L929 cells (repeated three times), and cell clones stably expressing murine CXCR3 molecule were screened by zeocin(500 mg/L). We used FCM and RT-PCR to verify expression of CXCR3 from protein level and gene level, respectively. Studied migration ability of L929-mCXCR3 interacted with its ligand IP-10 by transwell system. RESULTS: We have constructed recombinant murine CXCR3 gene retroviral vector and obtained L929-mCXCR3 gene transfected cell line which can stably expressing murine CXCR3 molecule. Positive expression rate of membrane is 97.0%, and it can directly migrate induced by IP-10, the chemotatic index is 4.356%. CONCLUSION: Construction of L929-mCXCR3 cell line has laid a good foundation on research of biologic characteristics of CXCR3 signal path , establishment of tumor metastasis model and preparation of anti-murineCXCR3 monoclonal antibody.

[Inhibitory and lethal effects of human-mouse chimeric antibody against CD80 on the growth of B lymphoma cell lines Raji and Daudi].

AIM: To study the inhibitory and lethal effects of human-mouse chimeric antibody against CD80 (named ch-4E5) on the growth of B lymphoma cell lines Daudi and Raji. METHODS: Immunofluorescence and flow cytometry were used to analyze the detection of membrane CD80 in Raji and Daudi by ch-4E5. After the co-culture of ch-4E5 with Raji and Daudi, respectively, at the final concentration of 10 mg/L, the expression of Fas and FasL was observated to be at the 0 h, 4 h, 10 h, 16 h, 24 h and 48 h by direct immunofluorescence and flow cytometry, and the blocking effect on cell growth of ch-4E5 was determined at 72 h by MTT assay. MTT assay was also used to study the ADCC effect with PBMC as effector cells and Raji and Daudi as target cells. The efficiency target ratio was 20:1. RESULTS: The combination rate between ch-4E5 and Raji and Daudi was 98.6% and 96.4%, respectively. After the co-culture of ch-4E5 with Raji and Daudi cells for 4 hs, the expression of FasL in Raji began to up-regulate. It reached the peak at 16 h and its positive rate was 16.8%. Compared with human IgG1 control group, it was increased obviously (P<0.01). The expression of Fas increased at 10 h, and then reached the top at 24 h. The combination rate was 15.6%. There was significant deviation compared with human IgG1 control group (P<0.01). Moreover, the expression of FasL and Fas on Daudi was also altered. The trend was similar to these on Raji, and the highest expression was 15.9% and 13.7%, respectively. The inhibitory rate was 34.60% and 32.64% respectively when ch-4E5 with Raji and Daudi had been co-cultured for 72 h (P<0.01). Furthermore, ch-4E5 could mediate the ADCC effect and the maximum killing rate was 55.61% and 54.42%, respectively(P<0.01). CONCLUSION: The human-mouse chimeric antibody against CD80 can inhibit the proliferation of B lymphoma cell lines Daudi and Raji cells through Fab and Fc sections in vitro.

[Expressions of urotensin II and its receptor in pulmonary arteries in rats with chronic thromboembolic pulmonary hypertension].

OBJECTIVE: To investigate the expressions of Urotensin II (UII) protein and mRNA and its receptor (UT) mRNA of medium and small pulmonary arteries of rats with chronic thromboembolic pulmonary hypertension. METHODS: The Wistar rats were injected thrombi through the jugular vein 2 times in 2 weeks and tranexamic acid was injected peritoneally once daily during the experiment to prevent thrombolysis. The mean pulmonary artery pressure (mPAP) was measured using right cardiac atheterzation. The expressions of UII protein in pulmonary arteries were studied by immunohischemistry with a polycolonal antibody. The expressions of UII mRNA and UT mRNA were detected by in situ hybridization using UII and UT oligonuclear probes. The changes of structures in pulmonary vessle were observed, including relative medial thickness of pulmonary artery (PAMT) and vessle wall area/total vessle area (WA/TA). RESULTS: The mPAP of the 4 weeks to the 12 weeks groups were (19.9 +/- 6.2) mm Hg (1 mm Hg = 0.133 kPa), (23.8 +/- 4.1) mm Hg and (27.4 +/- 5.4) mm Hg, higher than that of the control group (F = 13.75, P < 0.01, respectively). The PAMT of the 4 weeks to the 12 weeks groups were (42.6 +/- 11.16)%, (47.82 +/- 10.02)% and (53.79 +/- 10.41)%, and WA/TA of the 4 weeks to the 12 weeks groups were (22.75 +/- 6.79)%, (25.32 +/- 4.90)% and (27.05 +/- 7.71)%, both changed significantly as compared to the control group (F = 5.52 and 6.61, P < 0.01, respectively; P < 0.05 in 4 weeks group; P < 0.01 in 8 weeks and 12 weeks groups, respectively). The expressions of UIIprotein, UII mRNA and UT mRNA in the 4 weeks to the 12 weeks groups were obviously higher than the control group (F = 30.39, 30.78 and 14.49, P < 0.01, respectively), and their expressions were more marked in the small pulmonary arteries than in medium pulmonary arteries. The expressions of UIIprotein, UII mRNA and UT mRNA were positively correlated with mPAP and PAMT. The pulmonary vascular remodeling was time-dependently aggravated after embolism (r: 0.822, 0.866 and 0.846; 0.675, 0.712 and 0.756, P < 0.01, respectively). CONCLUSIONS: The expressions of UII protein, UII mRNA and UT mRNA of pulmonary arteries in the animal models were higher than those in the control group. These dynamic changes of UII mRNA, UIIprotein and UT mRNA may contribute to the development of pulmonary hypertension and vascular remodeling after pulmonary thromboembolism.