BCS1Neonatal growth retardation and lactic acidosis initiated by novel mutation sites in L gene / 中华预防医学杂志

This study aims to analyze the clinical characteristics and genetic variations of two cases with developmental delay and lactic acidosis in a family, and to explore the relationship between genetic variations and clinical features. A retrospective analysis was conducted on the clinical characteristics of two siblings with developmental delay and lactic acidosis who were treated at the Neonatal Department of Children's Hospital of Chongqing Medical University in May 2019 and December 2021, respectively. Whole-exome sequencing was used to detect genetic variations in the affected children. Homology modeling of the BCS1L protein was performed to analyze the structural and functional changes of the protein. The correlation between genetic variations and clinical phenotypes was analyzed. The results showed that the main clinical features of the two affected children in this family were manifestations of mitochondrial respiratory chain complex Ⅲ deficiency, including prematurity, developmental delay, respiratory failure, lactic acidosis, cholestasis, liver dysfunction, renal tubular lesions, coagulation dysfunction, anemia, hypoglycemia, hypotonia, and early death. Whole-exome sequencing revealed a novel deletion mutation c.486_488delGGA (p.E163del) and a novel missense mutation c.992C>T (p.T331I) in the BCS1L gene. Structural analysis of the homology modeling showed that the compound heterozygous mutation had a significant impact on protein function. In conclusion, the novel mutation site c.992C>T (p.T331I) in the BCS1L gene is a "likely pathogenic" mutation, and the compound heterozygous mutation is closely related to the phenotype of mitochondrial respiratory chain complex Ⅲ deficiency.

Mechanism of Dahuang Zhechongwan in Inhibiting Silicosis in Mice via p38 MAPK/NF-κB/TGF-β1 Pathway / 中国实验方剂学杂志

Objective:To explore the effects and mechanism of Chinese classical prescription Dahuang Zhechongwan on silicosis in mice. Method:Thirty-six male Kunming mice of SPF grade were randomized into the normal control group， model control group， tetrandrine （Tet， 0.039 mg·kg^-1） group， as well as high- （1.560 g·kg^-1）， medium- （0.780 g·kg^-1）， and low-dose （0.390 g·kg^-1） Dahuang Zhechongwan groups， with six mice in each group. Mice in all groups except for the normal control group underwent static inhalation of silica （SiO₂） dust for 40 consecutive days to induce fibrosis. After 28 days of intervention with corresponding drugs， the mice were sacrificed to collect the serum and lung tissues， with the former used for detecting tumor necrosis factor-<italic>α</italic> （TNF-<italic>α</italic>）， interleukin-1<italic>β </italic>（IL-1<italic>β</italic>）， IL-6， and hydroxyproline （HYP） levels by enzyme-linked immunosorbent assay （ELISA） and the latter for observing the pathological changes. Meanwhile， the protein and mRNA expression levels of p38 mitogen-activated protein kinase （p38 MAPK）， nuclear transcription factor-<italic>κ</italic>B （NF-<italic>κ</italic>B）， transforming growth factor-<italic>β</italic>₁ （TGF-<italic>β</italic>₁）， <italic>α</italic>-smooth muscle actin （<italic>α</italic>-SMA）， Smad2， Smad3， and Smad7 in the lung tissues were determined by Western blot and real-time polymerase chain reaction （Real-time PCR）. Result:Compared with the normal group， the contents of TNF-<italic>α</italic>， IL-1<italic>β</italic>， IL-6 and HYP in the model group were significantly increased， the difference was statistically significant（<italic>P</italic><0.05，<italic>P</italic><0.01）； compared with the model group， the high-dose group of Dahuang Zhechongwan could significantly reduce the contents of TNF-<italic>α</italic>， IL-6 and HYP in the serum of mice（<italic>P</italic><0.05， <italic>P</italic><0.01）， indicating that Dahuang Zhechongwan could reduce the lung inflammation of silicosis mice. At the same time， compared with the normal group， the protein and mRNA expression levels of p38 MAPK， NF-<italic>κ</italic>B p65， TGF-<italic>β</italic>₁， <italic>α</italic>-SMA， Smad2 and Smad3 in the model group were significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）， while the protein and mRNA expression levels of Smad7 were significantly decreased（<italic>P</italic><0.01）； compared with the model group， the protein and mRNA expression levels of p38 MAPK， NF-<italic>κ</italic>B p65， TGF-<italic>β</italic>₁， <italic>α</italic>-SMA， Smad2 and Smad3 in the high-dose Dahuang Zhechongwan group were significantly increased the protein and mRNA expression levels were significantly decreased（<italic>P</italic><0.05，<italic>P</italic><0.01）， while Smad7 protein and mRNA expression levels were significantly increased（<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:Dahuang Zhechongwan ameliorates the alveolar inflammation， extracellular matrix （ECM） deposition， and fibrosis in mice with silicosis possibly by regulating the p38 MAPK/NF-<italic>κ</italic>B/TGF-<italic>β</italic>₁ pathway.

Mechanism of Dahuang Zhechongwan Against Multiple Organ Fibrosis： A Review / 中国实验方剂学杂志

Dahuang Zhechongwan （DHZCW） is a classic prescription from the Jingui Yaolue（《金匮要略》） by ZHANG Zhong-jing，with the effects of tonifying deficiency， relaxing the middle， promoting regeneration， and resolving stasis. It has been widely used in the clinical treatment of various diseases with definite efficacy achieved. The research on multiple organ fibrosis has shown that DHZCW can slow down the development of organ fibrosis in the heart， liver， kidney， lung， etc.， and good results in both clinical practice and experimental research have been obtained. The present study reviewed the previous investigations on the experimental mechanism of DHZCW in the treatment of multiple organ fibrosis and revealed that the pathogenesis was closely related despite different disease sites. From the perspective of traditional Chinese medicine （TCM），these diseases shared a common pathogenesis，which was manifested by deficiency. Long-term diseases led to the formation of "dried blood". From the perspective of modern medicine， the diseases all showed pathological changes in the deposition of extracellular matrix （ECM）， and their occurrence and development were all based on certain effector cells ［such as hepatic stellate cell （HSC） and pancreatic stellate cell （PSC）］， with same cytokines ［such as tumor necrosis factor-α （TNF-α），interleukin-6 （IL-6），IL-1β，and α-smooth muscle actin （α-SMA）］ and some key pathways ［transforming growth factor-β1 （TGF-β1）/Smad， phosphatidylinositol-3-kinase （PI3K）/protein kinase B （Akt）， and lipopolysaccharide （LPS） paracrine and autocrine mechanisms］ involved. As a classic prescription for "deficiency-induced dry blood"， DHZCW was effective in treating fibrosis， which was presumedly related to the inhibition of ECM deposition by intervening in the above-mentioned mechanisms， thereby delaying the disease progression. This study is expected to provide literature support to clarify the scientific connotation of DHZCW in the treatment of multiple organ fibrosis and lay a foundation for further experimental and clinical research.

An exploration into mechanism of leukocyte elevation activity of Lvjiao Buxue granules based on network pharmacology / 药学学报

The mechanism of leukocyte elevation activity of Lvjiao Buxue granules was studied by establishing the active components-targets network and protein interactions network and analyzing the functions and pathways of targets. The main active ingredients of Lvjiao Buxue granules were obtained by TCMSP and literature excavation. Based on the DRAR-CPI, GeneCards and CoolGeN, the active components of Lvjiao Buxue granules were predicted and screened. Cytoscape software was used to construct the drug-active components-target network, and a protein database was constructed by using String database and Cytoscape software. The relation of the main active ingredients and targets were validated by Systems Dock Web Site. The GO and KEGG pathways involved in the targets were analyzed by DAVID databases. Using DisGeNET database to attribute the type of targets. The results showed that 49 active components and 89 targets of Lvjiao Buxue granules were involved. The network results showed that the composition of purine ribonucleosides, the regulation of cell death, especially the biological processes such as neutrophil and oxidative stress were mainly involved in the regulation of metabolic, cancer, tuberculosis, PI3K-Akt signaling and many other pathways to play its elevating leukocytes effect. This study reflects the characteristics of multi-components-multi-targets and multi-pathways of Lvjiao Buxue granules, which laid the foundation for further research into the mechanism of leukocyte elevation activity of Lvjiao Buxue granules.

Research progress in pathogenesis of hemolytic anemia and its treatment with traditional Chinese medicine / 中国中药杂志

Hemolytic anemia is a common clinical disease with diverse pathogenesis. In recent years, the incidence of hemolytic anemia is increasing dramatically. The present clinical treatment of immunosuppressive agents or splenectomy is effective to some extent; however, the accompanied clinical adverse reactions are also significant. Traditional Chinese medicine (TCM) has beneficial therapeutic effect on hemolytic anemia, with the obvious advantages including curative effect, less adverse reactions, and low price. The pathogenesis of hemolytic anemia as well as the pharmacological effects and mechanisms of the compound, single herb, and monomer composition of TCM in the treatment of hemolytic anemia were reviewed, aiming to provide the basis for the clinical treatment of hemolytic anemia and the modern research on the mechanism of TCM.

Investigation on positive effect and possible mechanisms of Lvjiao Buxue Granules treatment on cyclophosphamide-induced leucopenia mice through 1H-NMR based metabolomics approach / 中草药

Objective To investigate the positive effect and possible mechanism of Lvjiao Buxue Granules on the leukopenia induced by cyclophosphamide in mice using 1H-NMR metabolomics approach. Methods Cyclophosphamide was used to establish the leucopenia mice, and the content of white blood cells, red blood cells, hemoglobin, platelets and other routine blood indexes were measured. The changes of endogenous metabolites in serum were analyzed by 1H-NMR metabolomics to investigate the regulation effect of Lvjiao Buxue Granules on serum endogenous metabolites in mice with leukopenia and the regulation mechanism of metabolic pathway. Results Compared with the control group, the contents of white blood cells, red blood cells, hemoglobin, and platelets in the model group were decreased significantly (P < 0.01), while the level of white blood cells and platelets in the treatment group were increased significantly (P < 0.05), and Lvjiao Buxue Granules can call back the level of potential differences metabolites. Conclusion Lvjiao Buxue Granules can improve leukopenia by regulating the immune function, energy metabolism, and amino acid metabolism.

Comparative study of the corticosterone and glutamate induced PC12 cells depression model by 1H NMR metabolomics / 药学学报

This study was designed to analyze the change of metabolites in the PC12 cells and its medium induced by corticosterone (CORT) and glutamate (Glu) by proton nuclear magnetic resonance (1H NMR) metabolomics. The multivariate statistical analysis was employed to identify the difference between control groups and induced groups, respectively. In addition, metabolite pathway analysis was performed to explore the characteristic of CORT-induced and or Glu-induced PC12 cells depression model, and to provide the references for the selection of in vitro depression models as well as the further understanding of the mechanism on depressive disorders. We found 36 differential metabolites in CORT-induced PC12 cells and medium and 42 in Glu-induced PC12 cells. Furthermore, correlation analysis results show that serine and 2-oxoisoleucine were associated with most differential metabolites in CORT-induced PC12 cells. Lactate and glutathione were significantly correlated to the vast majority of differential metabolites in Glu-induced PC12 cells. We speculated that CORT-induced PC12 cell models may affect the fatty acid metabolism and cell membrane structure, and Glu-induced PC12 cell models may have a difference in the glycolysis and antioxidants.

A study on the influence of medical insurance paying proportion to hierarchical diagnosis and treatment / 预防医学

Objective To analyze the influence of medical insurance paying proportion to hierarchical diagnosis and treatment, and to provide basis for the improvement of hierarchical diagnosis and treatment. Methods Data about average number of visits of insured residents in 2015 were obtained from Hangzhou medical insurance service database. Differences between visits to Community Health Service Centers (CHSC) of residents having Medical Insurance of Urban and Rural Residents (MIURR) and residents having Urban Employees' Basic Medical Insurance (UEBMI) were compared. And differences stratified by signed contracted services offered by General Practitioners (GPs) or not were further analyzed. Results Proportion of visits to CHSCs in all visits to medical institutions in residents with MIURR was 77.22%, which was significantly higher than the proportion in residents with UEBMI was 51.26%. In the residents with UEBMI, and the proportion of visits to CHSCs in all visits to medical institutions of the group sighed contracted services offered by GPs was significantly higher than the group non-sighed. Conclusion Increasing the gap between reimbursement ratio of CHSCs and higher medical institutions plays an important role in guiding health-seeking behavior of residents. Reformation of medical insurance payment and contracted services will guide residents to seek medical treatment and effective referral.

Efficacy and safety of Willis covered stent for treatment of blood blister-like aneurysm / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the safety and short-term efficacy of Willis covered stent for treatment of blood blister-like aneurysms (BBA).</p><p><b>METHODS</b>Eight patients with BBA were treated with Willis covered stent system during the period from December 2014 to February 2016. The guiding catheter was placed as high as possible to facilitate the delivery of the covered stent system.</p><p><b>RESULTS</b>s Nine covered stents were implanted in the aneurysms of 8 patients (8 aneurysms), and 8 stents were released successfully in the parent arteries. In 6 patients, angiography immediately after stent release showed complete disappearance of the aneurysm and the parent arteries remained patent. One patient experienced a minor endoleak after stent implantation, and another stent was implanted to eliminate the endoleak. Iatrogeniccarotid-cavernous fistula occurred in 1 patient due to tortuosity of the parent artery, for which superficial temporal artery-to-middle cerebral artery bypass combined with parent artery occlusion was performed instead; the patient recovered smoothly and the bypass remained patent at 6 months after the operation. No other periprocedural complications occurred in these patients. Follow-up study showed no new-onset neurological deficits in these 8 patients, who had mRS score of 0 in 6 patients and of 1 in 2 patients. Digital subtractive angiography at 6 months after the operation demonstrated no aneurysm in these patients, and only one patient showed mild stenosis in the parent artery.</p><p><b>CONCLUSION</b>Willis covered stents are effective for treatment of BBA with good safety and short-term outcomes.</p>

Onyx embolization for treatment of dural arteriovenous fistula: comparison of long- distance versus routine injection method / 南方医科大学学报

<p><b>OBJECTIVE</b>To compare the efficacy, clinical characteristics, safety, injection time and radiation exposure of Onyx embolization using a long-distance injection method and routine injection method for management of dural arteriovenous fistula (DAVF).</p><p><b>METHODS</b>The clinical data were retrospectively analyzed in 59 patients with DAVF treated with Onyx embolization using long-distance injection method (28 patients) and routine injection method (31 patients). The efficacy, safety, injection time and radiation exposure during Onyx embolization were compared between the two injections methods.</p><p><b>RESULTS</b>The average radiation dose exposure to the surgeon per procedure was significantly lower in the long-distance injection group than in the routine group. The injection time (P=0.53), injection volume (P=0.78), number of supply arteries (P=0.80), Cognard types (P=0.67), and effect of embolization (P=0.88) were all similar between the two groups.</p><p><b>CONCLUSION</b>Endovaseular treatment of intracranial DAVF with Onyx embolization using the long-distance injection method is feasible, safe and effective and can reduce the radiation exposure to the surgeon.</p>

RITA combined with temozolomide inhibits the proliferation of human glioblastoma U87 cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of RITA, a small molecule that targets p53, combined with temozolomide (TMZ) on proliferation, colony formation and apoptosis of human glioblastoma U87 cells and explore the underlying mechanism.</p><p><b>METHODS</b>Cultured U87 cells were treated with RITA (1, 5, 10, 20 µmol/L), TMZ, or RITA+TMZ (half dose) for 24, 48 or 72 h. MTS assay were used to detect the cell proliferation, and the cell proliferation rate and inhibitory rate were calculated. The effect of combined treatments was evaluated by the q value. The expressions of p53, p21 and other apoptosis-associated genes were detected by qRT-PCR and Western blotting; cell apoptosis was assayed using flow cytometry with Annexin V/PI double staining; colony formation of the cells was detected with crystal violet staining.</p><p><b>RESULTS</b>MTS assay showed that RITA at the 4 doses more potently inhibited U87 cell viability than TMZ at 72 h (P=0.000) with inhibitory rates of 25.94%-41.38% and 3.84%-8.20%, respectively. RITA combined with TMZ caused a more significant inhibition of U87 cells (29.21%-52.11%) than RITA (P<0.01) and TMZ (P=0.000) alone. At the doses above 5 µmol/L, the combined treatments with RITA+TMZ for 48 h resulted in q values exceeding 1.2 and showed an obvious synergistic effect of the drugs. Both RITA and TMZ, especially the latter, significantly increased the expressions of p53, p21, puma, and other apoptosis-associated genes to accelerate apoptosis and inhibit the growth and colony formation of U87 cells, and the effect was more obvious with a combined treatment.</p><p><b>CONCLUSION</b>RITA inhibits the growth of human glioblastoma cells and enhance their sensitivity to TMZ by up-regulating p53 expression, and when combined, RITA and TMZ show a synergistic effect to cause a stronger cell inhibition.</p>

Advances in study on baicalein for regulation of signaling pathways in myocardial ischemia/hypoxia / 中草药

Myocardial ischemia/hypoxia often results in damaging the structure and function of heart. Baicalein, a widely distributed flavonoid, has been shown to play an important role in the cardioprotection of ischemia/hypoxia in recent years. The cardioprotection of baicalein was mediated by regulating the cell signaling pathways and controlling the phosphorylation of key signaling molecules. These signaling pathways involved in phosphatidylinositol-3-kinase-serine/threonine kinase (PI3K/Akt), mitogen-activated protein kinase (MAPKs), and nuclear factor-κB (NF-κB), and their down signaling proteins HMGB1, MMP-2/-9, and eNOS, and apoptosis proteins as well. The interaction mechanism of baicalein on signaling pathway in myocardial ischemia/hypoxia has been reviewed.

Function and mechanism of neurotensin (NTS) and its receptor 1 (NTSR1) in occurrence and development of tumors / 中国中药杂志

As a neuropeptide, neurotensin (NTS) is widely expressed in central and peripheral nervous system, which is mainly mediated byneurotensin receptor1 (NTSR1) to activate the related downstream signaling pathways. After summarized the function and mechanism of NTS/NTSR1 in various malignant tumors, we found that NTS/NTSR1 played essential roles during tumor initiation and development. NTS/NTSR1 regulates tumor initiation, proliferation, apoptosis, metastasis and differentiation mainly through three pathways, including IP3/Ca2+ /PKC/MAPKs pathway, MMPs/EGFR/MAPKs (PI3K/Akt) pathway, or Rho-GTPsaes and non-receptor tyrosine kinase pathway. Besides, NTS/NTSR1 is also regulated by some upstream pathways and some traditional Chinese medicine preparations and traditional Chinese medicine therapies. In this article, we summarized the function of NTS/NTSR1 and its mechanisms, and discussed the prospective in its application to clinical diagnosis and drugs targeting.

Diagnosis and treatment of multiple intracranial aneurysms / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the diagnosis and treatment strategy of multiple intracranial aneurysms (MIA).</p><p><b>METHODS</b>We retrospectively analyzed 96 patients with MIA (234 aneurysms). The rupture site was determined on the basis of computed tomographic and angiographic findings, and the supposed ruptured aneurysm was treated with coiling OR clipping. All the patients' records were reviewed including all computed tomographic scans and angiograms.</p><p><b>RESULTS</b>Twelve patients received conservative treatment, 56 patients were treated by endovascular embolization, and 28 patients received clipping; 44 patients received one-stage treatment, and 4 patients needed a second therapy. In 36 patients, only the ruptured aneurysm was eliminated. The clinical outcomes of these 84 patients evaluated by Glasgow Outcome Scale grades were: absence of deficits in 62 patients, minor deficits in 12 patients, major deficit in 8 patients; death occurred in 2 cases. Thirty patients were available for a 6-month follow-up with DSA, which revealed stable occlusion of the aneurysms in 29 patients and the need of a retreatment due to recanalization in only one patient.</p><p><b>CONCLUSION</b>Correct localization of the rupture aneurysm based on a comprehensive diagnosis is key to MIA treatment. All the aneurysms should be treated in one session whenever possible to protect the patient from rebleeding.</p>

A study on the relationship between mental health status and medication adherence in tuberculosis patients / 预防医学

Objective To evaluate the prevalence of depression,anxiety and suicide behavior in patients suffering from tuberculosis in Hangzhou and to explore their relationship with medication adherence. Methods Demographic characteristics,self-rating anxiety scale (SAS),the center for epidemiological studies -depression (CESD),social support rating scale (SSRS),suicide behavior information and the morisky medication adherence scale (MMAS)were investigated in 973 tuberculosis patients who were selected by systematic random sampling.Results The means of SAS and CESD were 39.71 ±8.30 and 14.16 ±10.77 respectively,which were both higher than the norms(P<0.01).Totally 102 (10.48%)patients had anxiety and 333 (34.22%)were depressed.Out of 973 patients,60 (6.17%)reported suicide ideation after tuberculosis diagnosis.The prevalence of non -adherence was 20.55%,which was defined with MMAS score above one and more.The non -adherence group had higher anxiety,depression and suicide ideation prevalence than the adherence group (15.50%vs.9.18%,46.50%vs.30.66%,11.00%vs.4.92%respectively,P<0.01).The mean score of SSRS,subjective support,objective support and utilization of support in the non-adherence group were 30.71 ±5.15,4.61 ±2.07,19.74 ±4.55 and 6.34 ±1.93 respectively,which were 34.06 ±7.39,6.62 ± 2.27,20.67 ±5.27 and 6.77 ±2.23 in the adherence group respectively.SSRS and its three dimension scores were significantly lower in the non-adherence group than that in the adherence group (P<0.01).Conclusion These findings show a quite serious situation of psychological problems of tuberculosis patients in Hangzhou and suggest psychological intervention should be included in adherence intervention.

Clinical and procedural predictors of no-reflow in patients with acute myocardial infarction after primary percutaneous coronary intervention / 世界急诊医学杂志（英文）

BACKGROUND:The treatment of acute myocardial infarction (AMI) is thought to restore antegrade blood flow in the infarct-related artery (IRA) and minimize ischemic damage to the myocardium as soon as possible. The present study aimed to identify possible clinical predictors for no-reflow in patients with AMI after primary percutaneous coronary intervention (PCI). METHODS:A total of 312 consecutive patients with AMI who had been treated from January 2008 to December 2010 at the Cardiology Department of East Hospital, Tongji University School of Medicine were enrolled in this study. Inclusion criteria were:(i) patients underwent successfully primary PCI within 12 hours after the appearance of symptoms; or (ii) patients with ischemic chest pain for more than 12 hours after a successful primary PCI within 24 hours after appearance of symptoms. Exculsion criteria were:(i) coronary artery spasm; (ii) diameter stenosis of the culprit lesion was ≤50％ and coronary blood flow was normal; (iii) patients with severe left main coronary or multivessel disease, who had to require emergency revascularization. According to thrombolysis in myocardial infarction (TIMI), the patients were divided into a reflow group and a no-reflow group. The clinical data, angiography findings and surgical data were compared between the two groups. Univariate and multivariate logistic regressions were used to determine the predictors for no-reflow. RESULTS:Fifty-four (17.3％) of the patients developed NR phenomenon after primary PCI. Univariate analysis showed that age, time from onset to reperfusion, systolic blood pressure (SBP) on admission, Killip class of myocardial infarction, intra-aortic balloon pump (IABP) use before primary PCI, TIMI flow grade before primary PCI, type of occlusion, thrombus burden on baseline angiography, target lesion length, reference luminal diameter and method of reperfusion were correlated with no-reflow (P<0.05 for all). Multiple logistic regression analysis identified that age >65 years [OR=1.470, 95％ confidence interval (CI) 1.460–1.490,P=0.007], long time from onset to reperfusion >6 hours (OR=1.270, 95％CI 1.160–1.400,P=0.001), low SBP on admission <100 mmHg (OR=1.910, 95％CI 1.018–3.896,P=0.004), IABP use before PCI (OR= 1.949, 95％CI 1.168–3.253, P=0.011), low (≤1) TIMI flow grade before primary PCI (OR=1.100, 95％CI 1.080–1.250,P<0.001), high thrombus burden (OR=1.600, 95％CI 1.470–2.760,P=0.030), and long target lesion (OR=1.948, 95％CI 1.908–1.990,P=0.019) on angiography were independent predictors of no-reflow. CONCLUSION:The occurrence of no-reflow after primary PCI for acute myocardial infarction can predict clinical, angiographic and procedural features.

H2O2 promotes neutrophil adherence and injury of human umbilical vein endothelial cells / 生理学报

To explore the role of hydrogen peroxide (H2O2) in promoting polymorphonuclear neutrophils adherence and injury of human umbilical vein endothelial cells (HUVECs), the ordinary optical microscope and scanning electron microscopy were used to observe the adherence and injury after HUVECs co-cultured with neutrophils pretreated by extracellular H2O2 (HUVECs and neutrophils co-culture without H2O2 pretreatment as control), and the adhesion rates of neutrophils were measured through cell count test. The percentages of HUVECs expressing intercellular adhesion molecule 1 (ICAM-1) and Apo2.7 were detected by flow cytometry. After being cocultured with the neutrophils pretreated by extracellular H2O2, HUVECs showed obvious injury changes, such as round or oval shape, shortened or disappeared microvilli, and membrane structural damage; The adhesion rate of neutrophils was (57.74 ± 9.18)%, which was significantly higher than that in control [(23.12 ± 6.43)%, P < 0.01, n = 8]; The percentages of HUVECs expressing ICAM-1 and Apo2.7 were (44.69 ± 1.52)% and (39.29 ± 1.81)% respectively, which were significantly higher than those in control [(21.79 ± 1.43)% and (9.79 ± 1.43)%] (P < 0.01, n = 8). The results suggest that extracellular H2O2 can promote the neutrophils adherence and injury of HUVECs.

Prenatal exposure to lipopolysaccharide results in vascular renin-angiotensin system dysfunction in offspring rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the impact of prenatal exposure to lipopolysaccharide on renin-angiotensin system of offspring rats.</p><p><b>METHODS</b>Six pregnant SD rats were randomly divided into 2 groups. The rats in the lipopolysaccharide (LPS) group were treated with LPS 0.79 mg/kg (i.p.) on the 8th, 10th and 12th day of gestation, and rats in the control group were treated with saline at the same time points. The blood pressure of offspring rats was measured by the tail cuff method. Protein expression of Angiotensin II (AngII) in thoracic aorta vessel was determined by immunohistochemistry. Protein expressions of AngII type 1 and type 2 receptor in thoracic aorta vessel were detected by Western blot.</p><p><b>RESULTS</b>Blood pressure of 12-week-old offspring rats of LPS group was significantly higher than that of 12-week-old offspring rats of control group (P < 0.01). The protein expression of AngII and AngII type 1 receptor in thoracic aorta vessel were significantly higher while protein expression of AngII type 2 receptor was lower in 15-week-old offspring rats of LPS group than in control group, resulting in a significant increase in the ratio of AngII type 1 receptor/AngII type 2 receptor in the aorta at 15-week-old of offspring rats than in 15-week-old offspring rats of control group (P < 0.01).</p><p><b>CONCLUSION</b>Prenatal lipopolysaccharide exposure results vascular renin-angiotensin system dysfunction, which may play an important role on the pathogenesis of hypertension development in offspring rats.</p>

Construction of let-7a expression plasmid and its inhibitory effect on k-Ras protein in A549 lung cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To elucidate the role of let-7a-mediated gene regulation in the pathogenesis of lung cancer.</p><p><b>METHODS</b>Two template DNA sequences were designed based on hsa-let-7a sequence in miRBase database. The let-7a expression construct and a control plasmid, namely pGenesil-let-7a and pGenesil-control, respectively, were generated by cloning the annealed oligonucleotides into pGenesil-1 and then transfected into A549 cells, which were selected by G418 to establish the lung cancer cell lines stably expressing let-7a-GFP and control-GFP. The living cells were counted by MTT assay and cell growth curves were drawn to analyze the cell proliferation. The k-Ras mRNA level was assessed by semi-quantitative RT-PCR, and the expression of k-Ras protein was determined by Western blotting and immunocytochemistry.</p><p><b>RESULTS</b>The recombinant vectors were verified by sequencing. The cell growth curves indicated that the proliferation of the cells transfected with pGenesil- let-7a were inhibited significantly compared with that of cells transfected with pGenesil-control and A549 cells. Semi- quantitative RT-PCR analysis showed that the levels of k-Ras mRNA almost remained unchanged in cells with or without the treatments. Western blotting and immunocytochemistry demonstrated a significant decrease of k-Ras protein levels in cells transfected with pGenesil-let-7a, but not in cells transfected with pGenesil-control, when compared to A549 cells.</p><p><b>CONCLUSION</b>let-7a over-expression represses the expression of k-Ras protein and significantly inhibits the growth of lung cancer cells.</p>

Expression of angiotensin converting enzyme and angiotensin converting enzyme 2 gene in lung of paraquat poisoning rats and protection of sodium dimercaptopropane sulfonate / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the expression of angiotensin converting enzyme (ACE) and ACE2 Gene in lung of paraquat poisoning rats and the protection of sodium dimercaptopropane sulfonate (Na-DMPS).</p><p><b>METHODS</b>One hundred SD male rats were randomly equally divided into 4 groups:normal control group (10 rats), drug control group (40 rats), paraquat poisoning group (40 rats) and drug intervention group(40 rats). The paraquat poisoning and drug intervention group rats were injected intraperitoneally by paraquat (20 mg/kg). The rats in drug intervention group rats were protected by intraperitoneal injection with Na-DMPS (200 mg/kg) 15 min before exposure of paraquat. Behavioral changes of the rats and histological changes of lung tissues under light microscope were observed. And the expression of ACE and ACE2 mRNA in lung tissues of rats both in paraquat poisoned group and drug intervention group were measured by RT-PCR at different time of 6 h, 24 h, 3 and 7 d after poisoning.</p><p><b>RESULTS</b>The poisoning symptoms of shortness of breath, cramps appeared and deteriorated progressively in rats after paraquat exposure and the protection of NA-DMPS could delay and reduce these symptoms significantly. Histological appearance of disorganization of pulmonary capillary and alveolus, exudation in alveolar space, pulmonary edema, severe bleeding, and inflammatory cells infiltration were obvious in lungs of rats after paraquat poisoning, whereas the histological changes were extenuated by protection of NA-DMPS. As compared with normal control group (NC group), the expressions of ACE, ACE2 mRNA in lung tissue decreased, and the lowest level of ACE mRNA expressions appeared at 24 h (0.457 +/- 0.262), on 3 d (0.385 +/- 0.179) after Paraquat exposure (P < 0.05), while lowest level of ACE2 mRNA expressions appeared on 3 d (0.415 +/- 0.247), 7 d (0.365 +/- 0.215) (P < 0.05). As compared with paraquat poisoned group, the expressions of ACE mRNA in lung tissue of rats in NA-DMPS protected group increased significantly at 24 h (0.739 +/- 0.558) and 3 d (0.749 +/- 0.414) (P < 0.05), while the expressions of ACE2 mRNA increased markedly on 3 d (0.584 +/- 0.345) and 7 d (0.493 +/- 0.292) (P < 0.05). But the expression of ACEmRNA and ACE2 mRNA in lungs had no statistical significance between normal control group and drug intervention group (P > 0.05).</p><p><b>CONCLUSION</b>The expressions of ACE and ACE2 mRNA in lung tissue of the rats with paraquat poisoning are decreased. Na-DMPS can effectively improve the balance of RAS in local lung tissue and reduce the pathological changes of lung tissue, delay the poisoning symptoms and show protective effects for acute lung injury induced by paraquat.</p>