OBJECTIVE: To evaluate the effect of Xiangdan Injection on mRNA expression of the endothelial vaso-active factors of patients with coronary heart disease and blood stasis. METHODS: Fifty-six patients were randomly divided into two groups:twenty-eight patients were treated according to the therapeutic guide for coronary heart disease as the control group and 28 were given the same treatment plus Xiangdan Injection as the treated group. The expressions of ET-1 and eNOS mRNA were examined with RT-PCR before experiment and ten days later. RESULTS: The positive rate of eNOS mRNA of the treated group increased, while the positive rate of ET-1 mRNA of the treated group decreased after ten day's treatment, with significant differences as compared with that before the experiment. Xiangdan Injection up-regulated the eNOS mRNA expression and suppressed the ET-1 mRNA expression. Changes of expression were not observed in the control group. CONCLUSION: Xiangdan Injection improves the endothelial function of patients with coronary heart disease and blood stasis by regulating the expressions of ET-1 and eNOS mRNA.

Objective The unbalanced phenotype of pe-ripheral blood monocyte is closely related to the pathological progres-sion of pulmonary fibrosis .The present study was designed to address the dynamic changes of circulating monocyte subsets in the experimen-tal mouse model of pulmonary fibrosis , and explore the relationship of circulating monocyte subsets with pulmonary inflammation and fibro-sis. Methods A total of 100male C 57BL/6J mice were random-ized as control group and a bleomycin A 5 group to be treated with sterile saline and bleomycin A5 at 2 mg/kg, respectively.The mice were sacrificed on day 1, 3, 7, 14, and 21 after treatment.The inflammation score and collagen volume fraction ( CVF) of the lung tissue were obtained by HE and Masson staining .The total number and different types of cells in the bronchoalveolar lavage fluid ( BALF) were counted using the routine method .The mRNA expressions of collagens ⅠandⅢwere determined by real-time PCR, the content of hydroxyproline (HYP) assayed by the chloramine-T method, and the proportions of different monocyte subsets measured by flow cytometry . Results Compared with the saline control , the bleo-mycin A5 group showed significantly increases in the inflammation score at 3 and 7 days ( P<0 .01 ) , CVF at 14 and 21 days ( P<0.01), and the numbers of total cells and macrophages in BALF at 3-21 days, the count of neutrophils granulocytes at 1-3 days (P<0.01), The numbers of neutrophile granulocyles were significant higer than that in control groups on the 1st(9.086 ±1.268 vs 1.108 ±0.229), 3rd(5.551 ±0.511 vs 0.315 ±0.100) and 7th(8.093 ±0.922 vs 0.249 ±0.074)day.The mRNA expressions of collagens ⅠandⅢat 14 and 21 days (P<0.05), the content of HYP at 7-21 days (P<0.01), and the proportion of Ly6Chi mon-ocytes on day 1, which peaked on day 3 (P<0.01) and then decreased from day 14 to 21.The proportion of Ly6Chi monocytes was positively correlated with the inflammation score (P<0.000 1) and CVF of the lung tissue (P=0.001 3). Conclusion In the mouse model of bleomycin A5-induced pulmonary fibrosis, dynamic changes of circulating Ly6Chi and Ly6Clo monocyte subsets occurred in different pathophysiological stages .Compared with the pathological process of inflammatory infiltration , Ly6Chi circulating monocytes displayed a rapid response to tissue injury and inflammation .The increased proportion of Ly6Chi monocyte subsets might be closely re-lated with pulmonary inflammation and fibrosis .

AIM: To establish a cell line of stable silencing of P2X7 receptor (P2X7R) expression through short hairpin RNA ( shRNA)-mediated interference in murine RAW264.7 macrophages, and to investigate the proliferation and apoptosis in the cell line.METHODS:Stable silencing of P2X7 R gene in the RAW264.7 cells was achieved by re-combinant shRNA plasmid targeting murine P2X7 R gene via liposome mediated transfection, followed by G418 selection. The efficacy of plasmid transfection and P2X7 R silencing in G418 resistant cells was verified by immunofluorescent micros-copy and real-time PCR, respectively.The proliferative activity was analyzed by CCK-8 assay and EdU cell proliferation as-say.The cell cycle distribution and apoptosis were evaluated by flow cytometry.RESULTS:The expression of P2X7 R at mRNA and protein levels was down-regulated by 80% in shP2X7 R group compared with negative control ( NC) plasmid transfection.In addition, P2X7 R-silencing cells exhibited higher proliferative activity compared with NC and wild-type RAW264.7 cells (P<0.05).Compared with NC cells, P2X7R silencing resulted in an increase in the phagocytosis of the cells ( P<0.05) .CONCLUSION:A cell line RAW264.7 of stable silencing of P2X7 R expression was successfully es-tablished.P2X7 R gene silencing stimulates the proliferation, and changes phagocytic function in murine RAW264.7 macro-phages.

Objective To clearify the features of an outbreak of Guillain-Barré syndrome (GBS) from Shuangyang district of Changchun city, Jilin province, China.Methods Serum samples obtained were tested serologically for antibodies to chlamydia, coxsackie virus, cytomegalovirus, and hepatitis B virus.Stool samples obtained were cultivated to search Campylobacterjejuni.Electrudiagnostic studies were done 1 day after onset to 10 days.Results Twenty-two patients (12 males and 10 females, aged 15-71 years) were from Shuangyang district, a suburb of Changchun, during a 2-week period in June to July 2007.An illness in the preceding 4 weeks was reported in 17 patients, 16 gave a history of diarrhoea, 1 patient had symptoms consistent with an "upper respiratory infection" and 1 patient had both.Limb weakness were the earliest symptoms.Respiratory assistance was needed by 3 patients.Tendon reflexes were lost as weakness developed.Eleven patients had raised concentrations of protein in the cerebrospinal fluid ((0.74±0.32) g/L), and the averaged cell count was 3×106/L There were 3 kinds of antibodies of hepatitis B virus positive above 17 GBS patients.Electrodiagnostic studies showed severe reductions in motor evoked amplitudes from distal stimulation in 12 patients.Sensory action potentials were normal.Conclusions The distinctive epidemiologicai, clinical and electromyography characteristics of these patients suggest that the disorder, different from acute inflammatory demyelinating polyneuropathy, is concordant with acute motor axonal neurupathy.The features of Shuangyang GBS was similar to those previously reported in several aspects.

To express recombinant Ancylostoma caninum anticoagulant peptide-c2 (AcAPc2), a whole cDNA fragment encoding AcAPc2 was achieved by ligation- PCR and inserted into prokaryotic expression vector pTWIN1 for constructing the specific self-splicing prokaryotic expression vector, pTWIN1-AcAPc2; positive recombinants were transformed into E. coli ER2566 for expression research. The recombinant protein, AcAPc2-intein2-CBD, was soluble and expressed in E. coli ER2566 (about 30.1% fusion protein in total protein). AcAPc2-intein2-CBD was characterized to be 41 KD by SDS-PAGE and identified by Western-blot. The recombinant fusion protein was purified to a efficiently high degree by chitin affinity chromatography. After the process of specific self-splicing induced by beta-Mercaptoethanol, the target protein, AcAPc2, was obtained, characterized to be 21 KD by SDS-PAGE and migrated as a dimmer. Molecular weight of AcAPc2 conformed to native dimmer. Bio-information analysis indicated relationship between secondary construction of AcAPc2 and biologic function. These findings greatly facilitate the purification of AcAPc2 and are very important for the additional studies on its anti-coagulation mechanism and its clinical application as anti-coagulation medicine.
Animals ; Dogs ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genes, Helminth ; Genetic Vectors ; Helminth Proteins ; biosynthesis ; genetics ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; RNA Splicing ; Recombinant Fusion Proteins ; chemistry ; pharmacology

OBJECTIVETo screen the point mutation of the low-density lipoprotein receptor (LDL-R) gene in Chinese familial hypercholesterolemia (FH) patients, characterize the relationship between the genotype and the phenotype and discuss the molecular pathological mechanism of FH.

METHODSA patient with clinical phenotype of homozygous FH and her parents were investigated for mutations in the promoter and all eighteen exons of the LDL-R gene. Screening was carried out using Touch-down PCR and direct DNA sequencing; multiple alignment analysis by DNASIS 2.5 was used to find base alteration, and the LDL-R gene mutation database was searched to identify the alteration. In addition, the apolipoprotein B gene (apo B) was screened for known mutations (R3500Q) that cause familial defective apo B100 (FDB) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).

RESULTSTwo new heterozygous mutations in exons 4 and 9 of the LDL-R gene were identified in the proband (C122Y and T383I) as well as her parents. Both of the mutations have not been published in the LDL-R gene mutation database. No mutation of apo B100 (R3500Q) was observed.

CONCLUSIONTwo new mutations (C112Y and T383I) were found in the LDL-R gene, which may result in FH and may be particularly pathogenetic genotypes in Chinese people.

Adult ; Apolipoproteins B ; genetics ; Asian Continental Ancestry Group ; Child ; China ; Female ; Heterozygote ; Homozygote ; Humans ; Hyperlipoproteinemia Type II ; genetics ; Male ; Mutation ; Receptors, LDL ; genetics

Background: Cardiopulmonary resuscitation (CPR) strategies in COVID-19 patients differ from those in patients suffering from cardiogenic cardiac arrest. During CPR, both healthcare and non-healthcare workers who provide resuscitation are at risk of infection. The Working Group for Expert Consensus on Prevention and Cardiopulmonary Resuscitation for Cardiac Arrest in COVID-19 has developed this Chinese Expert Consensus to guide clinical practice of CPR in COVID-19 patients. Main recommendations: 1) A medical team should be assigned to evaluate severe and critical COVID-19 for early monitoring of cardiac-arrest warning signs. 2) Psychological counseling and treatment are highly recommended, since sympathetic and vagal abnormalities induced by psychological stress from the COVID-19 pandemic can induce cardiac arrest. 3) Healthcare workers should wear personal protective equipment (PPE). 4) Mouth-to-mouth ventilation should be avoided on patients suspected of having or diagnosed with COVID-19. 5) Hands-only chest compression and mechanical chest compression are recommended. 6) Tracheal-intubation procedures should be optimized and tracheal-intubation strategies should be implemented early. 7) CPR should be provided for 20-30 min. 8) Various factors should be taken into consideration such as the interests of patients and family members, ethics, transmission risks, and laws and regulations governing infectious disease control. Changes in management: The following changes or modifications to CPR strategy in COVID-19 patients are proposed: 1) Healthcare workers should wear PPE. 2) Hands-only chest compression and mechanical chest compression can be implemented to reduce or avoid the spread of viruses by aerosols. 3) Both the benefits to patients and the risk of infection should be considered. 4) Hhealthcare workers should be fully aware of and trained in CPR strategies and procedures specifically for patients with COVID-19.