[ ABSTRACT] AIM:To investigate the effect of microRNA-132 ( miR-132 ) transfection on the lipopolysaccharide ( LPS)-induced inflammation in rat alveolar macrophages.METHODS:The rat alveolar macrophage NR8383 cultured with-out pyrogen in vitro were divided into blank control group, negative control group and transfected group.The cells in the 3 groups were transfected with phosphate buffer solution ( PBS) , Lipofectamine 2000 and synthesized miR-132 mimic respec-tively.The cell proliferation was detected by Cell Counting Kit-8 ( CCK-8) assay.Real-time PCR was used to detect the ex-pression of miR-132 in the cells.After NR8383 cells were stimulated with LPS for 6 h, the NF-κB DNA-binding activity was measured by electrophoretic mobility shift assay ( EMSA) .The expression of tumor necrosis factor-α( TNF-α) and interleu-kin-6 (IL-6) in NR8383 cells was assayed by Western blotting.RESULTS: After transfection, the expression of miR-132 was significantly higher than that in blank control group and negative control group.The growth of NR8383 cells in transfect-ed group was significantly inhibited compared with blank control group and negative control group ( P<0.05 ) .After the cells were stimulated with LPS, the productions of NF-κB, TNF-αand IL-6 in transfected NR8383 cells were decreased com-pared with blank control group and negative control group (P<0.05).CONCLUSION:Transfection of alveolar macropha-ges with miR-132 significantly suppresses the cell growth, and inhibits inflammatory responses induced by LPS.

Child ; Humans ; Hypertension, Pulmonary ; complications ; Male ; Pulmonary Heart Disease ; complications ; Sleep Apnea, Obstructive ; complications

Brain Neoplasms ; diagnosis ; metabolism ; pathology ; Child ; Diagnosis, Differential ; Female ; Glioma ; pathology ; Humans ; Immunohistochemistry ; Magnetic Resonance Imaging ; Meningioma ; diagnosis ; metabolism ; pathology ; Mucin-1 ; metabolism ; Occipital Lobe ; Vimentin ; metabolism

The intracellular trafficking and subcellular distribution of exogenous gene is very important for gene delivery. A successful gene vehicle should overcome various barriers including endosomal membrane barriers to delivery gene to the target organelle. Traditional nonviral vehicle is unable to avoid endosomal pathway efficiently, so the efficiency of gene delivery is low and the application of gene drugs is limited. In order to achieve efficient nonviral gene delivery, a lot of researches based on endosomal escape have been carried out and some agents with the function of endsomal escape have been found. These agents facilitate the endsomal escape via various mechanisms, such as fusion into the lipid bilayer of endosomes, pore formation in the endosomal membrane, proton sponge effect and photochemical methods to rupture the endosomal membrane. In this review, various reported strategies for endsomal escape are described according to the escape mechanisms, and their applications in intracellular gene delivery are also discussed.

Studies have shown that women's menopause caused by permanent cessation of ovarian function is closely related to lipid metabolism disorders. Er-xian Decoction has been used in the clinical treatment for gynecological diseases and has a good effect on diseases related to reduced sex hormone function. In this study, metabolomics was performed on bilateral ovariectomized model rats within 12 weeks after modeling to mimic the physiological state of menopausal women in different menopausal stages and Er-xian Decoction dosed model rats. The results of liver oil red O staining sections showed lipid metabolic disorder of bilateral ovariectomized model rats and the regulating effects of Er-xian Decoction. 46 potential biomarkers (6 steroid hormones, 3 sphingolipids, 11 phospholipids and 26 glycerides) in plasma and 32 potential biomarkers (1 steroid hormones, 20 phospholipids and 11 glycerides) in liver were obtained based on lipidomics analysis. Then, we analyzed the differential metabolic pathways and construct the lipid metabolism network significantly regulated by Er-xian Decoction. The results provided valuable information for in-depth understanding of the gradual changes on lipid metabolism disorders under menopausal conditions and the characteristics and mechanisms of compound Er-xian Decoction's regulatory effects. The study complied with the procedures established by the Animal Experiment Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and passed the animal experiment ethics examine (No. 00000918).

The intracellular trafficking and subcellular distribution of exogenous gene is very important for gene delivery. A successful gene vehicle should overcome various barriers including endosomal membrane barriers to delivery gene to the target organelle. Traditional nonviral vehicle is unable to avoid endosomal pathway efficiently, so the efficiency of gene delivery is low and the application of gene drugs is limited. In order to achieve efficient nonviral gene delivery, a lot of researches based on endosomal escape have been carried out and some agents with the function of endsomal escape have been found. These agents facilitate the endsomal escape via various mechanisms, such as fusion into the lipid bilayer of endosomes, pore formation in the endosomal membrane, proton sponge effect and photochemical methods to rupture the endosomal membrane. In this review, various reported strategies for endsomal escape are described according to the escape mechanisms, and their applications in intracellular gene delivery are also discussed.
Cell Membrane ; metabolism ; Endosomes ; metabolism ; Gene Transfer Techniques ; Genetic Therapy ; Genetic Vectors ; Humans

The study was purposed to analyze the frequency and distribution of irregular antibodies in Shaoguan area. Screening 15 033 random donor antibodies in Shaoguan area by screening cells, polybrene and antiglobulin tests. The results indicated that the irregular antibodies were found in 42 samples. The frequency of irregular antibodies in female was higher than that in male (P < 0.001), and Rh blood group antibodies such as anti-D, anti-E, and anti-Ec C were common (47.6%). 2 samples of Le antibodies were failed to be found by polybrene test. 2 samples of irregular antibodies with titer 2 were undiscovered by screening test of 10 pooled samples. In conclusion, because of irregular antibodies resulting in hemolytic transfusion reaction, the investigation of frequency and distribution of irregular antibodies is very important for safe transfusion. Antibody screening must be done for female donors, and especially for massive plasma transfusion of patients with severe and dangerous illness and infants so as to ensure safety.
Adolescent ; Adult ; Blood Donors ; China ; Erythrocytes ; immunology ; Female ; Humans ; Isoantibodies ; blood ; Male ; Mass Screening ; Middle Aged ; Rh-Hr Blood-Group System ; blood ; immunology ; Rho(D) Immune Globulin

Objective To evaluate the value of CK34?E12,p63 and P504S immunostaining in diag- nosis of benign and malignant lesions of the prostate.Methods Expression of CK34?E12,p63 and P504S in 74 benign and malignant lesions of prostate,including 27 PC,6 HGPIN,10 LGPIN,3 AAH and 28 BPH were observed by immunohistochemical method.Results Expression of p63 and CK34?E12 were positive in AAH,LGPIN and BPH,all of PC were negative,the positive rate of HGPIN was 83.3%(5/6).There were sig- nificant differences in p63 and CK34?E12 expression between PC and AAH,HGPIN,LGPIN,BPH(P

To observe of alloantibodies of patients with autoimmune hemolytic anemia (AIHA), the alloantibodies masked by autoiantibody were detected by using chloroquine elution test, and the specificity of autoantibody was identified by ether elution test. The results showed that 19 cases out of 38 patients with AIHA were positive detected by indirect antiglobulin test and in 7 cases of them alloantibodies in sera cases were found (1 case of anti-D, 4 cases of anti-E and 2 cases of anti-CE), in 5 cases of them alloantibody were detected carried blood group specificity (3 cases of anti-E, anti-C and anti-c 1 case respectively). In conclusion, detections of alloantibodies by chloroquine elution test and ether elution test were very important for transfusion safety in therapy of patients with AIHA.
ABO Blood-Group System ; Adolescent ; Adult ; Anemia, Hemolytic, Autoimmune ; immunology ; Autoantibodies ; blood ; Blood Grouping and Crossmatching ; methods ; Erythrocytes ; immunology ; Female ; Humans ; Isoantibodies ; blood ; Male ; Middle Aged ; Rh-Hr Blood-Group System

OBJECTIVETo study the HBsAg transient expression in HepG2 or COS-7 cells with eukaryotic expression plasmids inserting HBsAg gene (pCI-S and pcDNA3.1-S) and the efficacy of naked DNA immunization in mice.

METHODSFirstly, the recombinant plasmids of pCI-S and pcDNA3.1-S were constructed by the cloning technique and the accuracy of these constructs was confirmed by restriction enzyme digestion and DNA sequencing. Secondly, plasmids of pCI-S and pcDNA3.1-S were transferred into HepG2 and COS-7 cells, respectively by means of cationic liposome. HBsAg transient expression was assayed by ELISA in cell culture supernatants and cell lysates. Thirdly, plasmids were injected into quadriceps muscles of BALB/C mice and serum samples were obtained from individual immunized or control mice 4 weeks after injection and boost injection, respectively. Anti-HBs were assayed in mice sera by ELISA. HBsAg-specific CTL responses of spleen cells from immunized mice were tested by the LDH method.

RESULTSPlasmids of pCI-S and pcDNA3.1-S allowed HBsAg transient expression in cell culture supernatants and cell lysates of HepG2 or COS-7 cells. Intramuscular immunization of BALB/C mice with plasmids of pCI-S or pcDNA3.1-S elicited the antibody and cytotoxic T lymphocyte responses to HBsAg.

CONCLUSIONSThe vectors used in this study are effective to induce prime antibody and HBsAg-specific-cytotoxic T lymphocyte responses to HBsAg in mice after intramuscular immunization.

Animals ; COS Cells ; Cloning, Molecular ; DNA, Viral ; genetics ; Eukaryotic Cells ; metabolism ; Female ; Gene Expression ; Hepatitis B ; immunology ; prevention & control ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis, Viral, Animal ; immunology ; prevention & control ; Humans ; Immunization ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection ; Tumor Cells, Cultured ; Vaccines, DNA ; genetics ; immunology ; Viral Vaccines ; genetics ; immunology