Endothelial METRNL determines circulating METRNL level and maintains endothelial function against atherosclerosis

METRNL is a recently identified secreted protein with emerging functions. This study is to find major cellular source of circulating METRNL and to determine METRNL novel function. Here, we show METRNL is abundant in human and mouse vascular endothelium and released by endothelial cells using endoplasmic reticulum-Golgi apparatus pathway. By creating endothelial cell-specific Metrnl knockout mice, combined with bone marrow transplantation to produce bone marrow-specific deletion of Metrnl, we demonstrate that most of circulating METRNL (approximately 75%) originates from the endothelial cells. Both endothelial and circulating METRNL decrease in atherosclerosis mice and patients. By generating endothelial cell-specific Metrnl knockout in apolipoprotein E-deficient mice, combined with bone marrow-specific deletion of Metrnl in apolipoprotein E-deficient mice, we further demonstrate that endothelial METRNL deficiency accelerates atherosclerosis. Mechanically, endothelial METRNL deficiency causes vascular endothelial dysfunction including vasodilation impairment via reducing eNOS phosphorylation at Ser1177 and inflammation activation via enhancing NFκB pathway, which promotes the susceptibility of atherosclerosis. Exogenous METRNL rescues METRNL deficiency induced endothelial dysfunction. These findings reveal that METRNL is a new endothelial substance not only determining the circulating METRNL level but also regulating endothelial function for vascular health and disease. METRNL is a therapeutic target against endothelial dysfunction and atherosclerosis.

One-Step Rapid Quantification of Serum Neutralizing Antibody after COVID-19 Vaccination by a High-Throughput Nanoplasmonic Sensor Platform

The COVID-19 vaccination efficacy depends on serum production level of the neutralizing IgG antibody (NA) specific to the receptor binding domain of SARS-Cov-2 spike protein. Therefore, a high-throughput rapid assay to measure the total SARS-CoV-2 NA level is urgently needed for COVID-19 serodiagnosis, convalescent plasma therapy, vaccine development, and assessment. Here, we developed a nanoplasmonic immunosorbent assay (NanoPISA) platform for one-step rapid quantification of SARS-CoV-2 NAs in clinical serum samples for high-throughput evaluation of COVID-19 vaccine effectiveness. The NanoPISA platform enhanced by the use of nanoporous hollow gold nanoparticle coupling was able to detect SARS-CoV-2 NAs with a limit of detection of 0.1 ng/mL within 15 min. The one-step NanoPISA for SARS-CoV-2 NA detection in clinical specimens yielded good results, comparable to those obtained in the gold standard seroneutralization test and the surrogate virus neutralizing ELISA. Collectively, our findings indicate that the one-step NanoPISA may offer a rapid and high-throughput NA quantification platform for evaluating the effectiveness of COVID-19 vaccines.

One-Step Rapid Quantification of SARS-CoV-2 Virus Particles via Low-Cost Nanoplasmonic Sensors in Generic Microplate Reader and Point-of-Care Device

The spread of SARS-CoV-2 virus in the ongoing global pandemics has led to infections of millions of people and losses of many lives. The rapid, accurate and convenient SARS-CoV-2 virus detection is crucial for controlling and stopping the pandemics. Diagnosis of patients in the early stage infection are so far limited to viral nucleic acid or antigen detection in human nasopharyngeal swab or saliva samples. Here we developed a method for rapid and direct optical measurement of SARS-CoV-2 virus particles in one step nearly without any sample preparation using a spike protein specific nanoplasmonic resonance sensor. We demonstrate that we can detect as few as 30 virus particles in one step within 15 minutes and can quantify the virus concentration linearly in the range of 103 vp/ml to 106 vp/ml. Measurements shown on both generic microplate reader and a handheld smartphone connected device suggest that our low-cost and rapid detection method may be adopted quickly under both regular clinical environment and resource-limited settings.

Establishment of mouse myocardial infarction model and early electrocardio- gram assessment / 药学实践杂志

Objective To establish and optimize a mouse myocardial infarction (MI) model, and to use twice limb lead ECGs immediately after coronary ligation and 4 h after surgery to evaluate the occurrence of myocardial infarction. Methods Twenty-nine male C57BL/6J mice were anesthetized with isoflurane. then a myocardial infarction model was established by ligating the left anterior descending (LAD) coronary artery through the third/fourth intercostal space of left anterior chest. Immediate and 4 h postoperative limb lead ECGs were performed. Twenty-four hours after surgery, the chest was opened and the occurrence of myocardial infarction was evaluated. The heart samples were taken for TTC staining to determine the infarct area and calculate the infarct area. Results During the mice underwent coronary artery ligation the intraoperative mortality was 6.8% (2/29), and the early postoperative (<4 h) mortality was 10.3% (3/29). The 24 h survival rate was 82.8% (24/29). 24 hours after TTC staining confirmed the occurrence of infarction, the myocardial infarction model was established. The success rate of the model was 79.3% (23/29), and the average infarct size (infarcted myocardial weight / whole ventricular weight) was (28 ± 6)%; The mice successfully established by the model showed obvious ST-T changes in the ECG at 4 hours after surgery, suggesting that a myocardial infarction has occurred. Conclusions The mouse myocardial infarction model was successfully established. The combined use of ECG immediately after surgery and 4 h after surgery could be used as a rapid and non-invasive evaluation method for mouse myocardial infarction.

Establishment and evaluation of in vivo and in vitro D-galactose induced cognitive impairment models / 药学实践杂志

Objective To construct and explore the in vivo and in vitro D-galactose induced cognitive impairment models and evaluate the application value of the combined models in the study of cognitive impairments.Methods The cognitive impairment mice model induced by D-gal was prepared by continuous intraperitoneal injection of D-gal saline solution for 8weeks, followed by detection of learning and memory functions with Morris water maze.The related molecular markers in the brain tissue were assayed to evaluate the effect and application value.D-gal cell model was prepared by adding D-gal in different concentrations into the cell cultural medium of neurons harvested from the hippocampus of young mice.The effect and application value were evaluated by detecting the molecular markers related to the level of cell injury.Results The Morris water maze on the D-gal model showed that the learning and memory functions of mice in the model group were significantly lower than those in the control group.Meanwhile, the levels of apoptosis and oxidative stress in the model group were significantly higher than those in the control group.In the hippocampal neuron model of D-gal, the neurons showed a dose-dependent morphologic and functional change with the increase of D-gal dose and the levels of apoptosis and oxidative stress were significantly higher than those in the negative control.Conclusion D-galactose can be successfully used to induce cognitive impairment models both in vivo and in vitro through the decrease of the learning and memory functions of mice and induction of apoptosis and oxidative stress in neurons.Combined application of the two models of D-gal can be one of effective and promising tools for the study of cognitive impairment and pharmacodynamic evaluation.

Clinical efficacy of improved TIPS therapy in portal hypertension with acute upper gastrointestinal bleeding / 实用放射学杂志

Objective To discuss the clinical efficacy of improved TIPS of percutaneous portal vein puncture in treatment of acute upper gastrointestinal bleeding induced by portal hypertension.Methods 28 patients with acute upper gastrointestinal bleeding underwent improved TIPS therapy in our hospital were enrolled.The clinical data,laboratory parameters and hemodynamic changes were collected and analyzed before and after operation.Results The success rate for the first time and hemostatic rate of postoperative 24 hours in all patients was 100%.2(7.14%)patients underwent mild hepatic encephalopathy.After TIPS operation,the concentration of serum albumin increased,whereas,concentration of total bilirubin and alanine aminotransferase decreased(P<0.01).Portal vein pressure (PVP)of pre-and post-operation was(41.48 ± 3.72)mmHg and(28.91 ± 2.59)mmHg,and the hepatic venous pressure gradient (HVPG)was(20.30 ± 2.76)mmHg and(8.81 ± 2.04)mmHg.PVP and HVPG were both decreased significantly after operation(P<0.01).Conclusion Improved TIPS therapy can obtain good clinical efficacy and safety for esophageal and gastric varicose bleeding in acute cirrhosis.

Quality of control of compound chlorhexidine acetate ointment / 药学实践杂志

Objective To improve the standard of the quality control of compound chlorhexidine acetate ointment . Methods TLC was used to control the quality of menthol crystal and camphor .A method to determine chlorhexidine acetate and cocaine hydrochloride simultaneously by HPLC was established .Results The spots of menthol crystal and camphor in TLC were clear .Chlorhexidine acetate and cocaine hydrochloride showed excellent linearity ,which were at the range of 10.01-160.14 μg/ml and 10 .01-160 .14 μg/ml ,respectively .The average recoveries were 101 .5% (RSD=1 .8% ) and 100 .5% (RSD=2 .8% ) .Conclusion The methods were simple ,sensitive and with good reproducibility and could be used to control the quality of compound chlorhexidine acetate ointment .

Improvement of quality control of lidocaine hydrochloride injection / 药学实践杂志

Objective To improve the quality control standard of lidocaine hydrochloride injection .Methods A method for determination of related substances in lidocaine hydrochloride injection was established .Lidocaine hydrochloride was as‐sayed by HPLC .The chromatographic conditions :C18 chromatographic column was used .The mobile phase was phosphate buffer and acetonitrile (50∶50 ,adjusted to pH 8 with phosphoric acid) .The detection wavelength was 254 nm .Results Ac‐cording to the result of method verification ,related substances could be examined by HPLC .Lidocaine hydrochloride was as‐sayed by HPLC ,which showed excellent linearity at the range of 373 .62‐3 736 .19 μg/ml .The average recoveries were 102 .1% (RSD=0 .9% ) .Conclusion The improved standard could be used to control the quality of lidocaine hydrochloride injection .

Determination of Compound Sodium Acetate Injection / 中国药师

Objective:To improve the determination method for compound sodium acetate injection. Methods:Atomic absorption spectrophotometry ( AAS) was used to determine sodium chloride, calcium chloride and potassium chloride in compound sodium acetate injection. Results:The linear range of calcium ion, potassium ion and sodium ion was 9.124 ×10 -7-1.369 ×10 -5 g·ml-1(r =0.999 5),1.501 ×10 -7 ~4.504 ×10 -6 g·ml-1(r=0.999 2) and 7.500 ×10 -8-2.251 ×10 -6 g·ml-1(r=0.999 5), and the av-erage recovery was 100. 4%(RSD=1. 4%,n=9),102. 4%(RSD=1. 6%,n=9) and 100. 3% (RSD=1. 1%,n=9), respectively. Conclusion:The method is simple, accurate and reproducible, and can be used to control the quality of compound sodium acetate in-jection.

The histological changes of diabetic rats' skin and the effects on the percutaneous absorption of glucocorticoid / 药学学报

To examine the histological changes of diabetic rats' skin and the effects on the percutaneous absorption of hydrocortisone (HC, a glucocorticoid), male Wistar rats were randomly divided into five groups: control group, diabetes one-week group (W1), two-week group (W2), three-week group (W3), and four-week group (W4), while each group contained 6 rats. Diabetes mellitus (DM) rat model was prepared with the method of streptozocin (STZ, 40 mg x kg(-1)) intraperitoneal injection. Abdominal skin was cut to carry out an in-vitro penetration experiment on an improved Franz diffusion cells, and phosphate buffer (PBS, pH 7.4) was used as receptor solution. The solution was analyzed with HPLC, and then the penetrating rate can be calculated. Meanwhile, rats' abdominal skins of different DM periods were HE stained and made into tissue slices to find if any histological changes occurred. The penetrating rate of control, W1, W2, W3, and W4 groups were 2.39 +/- 1.25, 3.22 +/- 1.72, 3.02 +/- 1.89, 3.63 +/- 2.02 and 5.00 +/- 3.36 microg x h(-1) x cm(-2), respectively. There was significant difference between the control and the W4 group (P 0.05). The tissue slices showed that compared to the normal rats' skin, little change was observed in one-week DM rats' skin, but the skin of one-month DM rats' skin was observed thinner, and it became much thinner than that of rats with two-month diabetes, especially the epidermis. After making a rat into diabetic, the rats' skin goes through a pathological change, and this change is closely interrelated with the increase of the permeation of HC. Therefore, it is necessary to adjust the dose while some drug was applied on the skin in case of diabetes mellitus.

Effect of mfn2 on mitochondrial function in steatosis hepatocytes / 中国病理生理杂志

AIM: To investigate the effect of mfn2 on mitochondrial function in steatosis hepatocytes. METHODS: Plasmid pEGFP-mfn2 was transfected into hepatocyte strain L02 by Lipofectamine 2000 in vitro, then the steatosis model of hepatocytes was establish by oleic acid induction. RT-PCR was used to evaluate mRNA expression and Western blotting was use to detect the protein expression. ATP level was determined by firefly luciferase bioluminescent. ROS production was measured by fluorescence probe DCFH-DA. Chondrosome transmembrane potential of L02 was observed by labeling of JC-1 and FCM. RESULTS: The stable expression of ectogenesis mitofusin2 in L02 cells was confirmed by RT-PCR and Western blotting. In the model of oleic acids induced lipid formation, Mfn2 obviously inhibited the descent of chondrosome transmembrane potential and ATP level, and increased ROS production in L02 cells. CONCLUSION: Up-regulated expression of mfn2 attenuates mitochondria dysfunction caused by oleic acids induced lipid formation.

Clinical study on Salvia injection (freeze-dried) in the interventiontreatment of the oviduct obstructive infertility / 中国介入影像与治疗学

Objective To study the clinical efficacy of the injection Salvia (freeze-dried, abbr: Salvia powder needle), with fallopian tube recanalization on the treatment of oviduct obstructive infertility. Methods One hundred and ninety-five patients of oviduct obstructive infertility were enrolled in this study and divided into drug group and control group. Salpingography with modified catheter, recanalization and perfusion operation were performed in both two groups. Salvia powder, gentamicin, α-ACT and dexamethasone were injected into test group, while Gentamicin, α-ACT and dexamethasone were injected into control group. Recanalization rate of fallopian tube, rate of intrauterine gestation and rate of ectopic pregancancy were measured and compared between two groups. Results Twelve months after operation, recanalization of fallopian tube of test group and control group was 94.39% and 85.87%, respectively. Twenty-four months after operation, intrauterine gestation rate of test group and control group was 50.00% and 35.71%, respectively. Conclusion Salvia powder combined with modified recanalization operation is helpful to improve the rate of intrauterine gestation.

Comparison of Bioavailability of Fleroxacin Tablets from Three Different Factories / 中国药房

0.05),and T max had signif?icant differences(P