Objective To investigate the effect of down-regulated Blimp1 gene expression on differenetiation of bone marrow cells into dendritic cells (DCs).Methods Blimp1-shRNA was constructed and then loaded into lentivirus vector as lenti-blimp1-shRNA.Bone marrow cells from Balb/c mice were induced differentiation to DCs in an 8-day cell culture system with GM-CSF/IL-4 incubation and LPS stimulation at day 7.The cells were divided into groups as empty control (no treatment),lenti-control (transfected by lentivirus empty vector at day 1),and lenti-Blimp (transfected by lenti-blimp1-shRNA at day 1).The transfection efficiency was evaluated by GFP fluorescence for one week.The morphology and growth curve were analyzed.Real-time PT-PCR and Western blotting were used to evaluate mRNA and protein expression of Blimp1.At day 8,CD11 c and CD86/MHC-Ⅱ were quatitified using flow cytometry.Results GFP fluorescence emerged 3 days after transfection and was continuously expressed.Classic DC morphology was shown in no treatment cells,while damaged morphology presented in the cells with lentivirus transfection.The empty control cells proliferated from day 3,peaked as (2.45 ± 0.26) 106/well at day 4,and kept at (2.27 ± 0.19) 106/ well at day 8,The cells receiving lentivirus presented (1.69 ± 0.39) 106/well.The expression of Blimp1 mRNA and protein in the lenti-Blimp1 group was 76％/1％ and 1.0％/74.0％ of the empty control group.At day 8,CD11c,CD86 and MHC-Ⅱ expression in the empty control group was (69.2 ±5.0)％,(51.1± 4.9) ％ and (56.3 ± 7.3) ％,while (68.6±5.9)％,(49.5±4.3)％ and (69.4±4.5)％ in the lenti-control group,and (72.8 ± 5.5)％,(50.2 ± 6.0)％ and (46.5 ± 5.7)％ in the lenti Blimp1 group.Conclusion Lentivirus-mediated Blimp1-shRNA gene therapy modulates blimp1 expression of DC precursors.Down-regulation of Blimp1 fails to interrupt the differentiation of DCs but inhibits the maturation.

Objective To investigate the effect of adenosine A2A receptor on pituitary-adrenal axis response in acute phase of moderate craniocerebral trauma.Methods Eighteen adenosine A2A receptor knock-out mice in a C57BL/6 background and another eighteen their wild-type littermates were divided into normal control group and craniocerebral trauma for 4 hours group,and craniocerebral trauma for 24 hours group according to random number table,with siμ mice per group.Plasma levels of adrenocorticotropic-hormone (ACTH) and corticosterone at hours 4 and 24 postinjury were determined using ELISA method.Results At 4 and 24 hours,brain water content in wild-type mice [(80.950 ± 0.184) ％,(82.178 ± 0.255)％ respectively] was higher than that in gene knock-out mice [(80.006 ± 0.199)％,(81.091 ± 0.295)％ respectively,P ＜ 0.01].Besides,brain water content in both wild-type and gene knock-out mice increased after injury (P ＜ 0.01).Plasma levels of ACTH and corticosterone were higher in geneknock-out sham mice than in wild-type sham mice [(120.214 ± 2.472) ng/L vs (91.767 ±7.395) ng/L,(27.814 ±0.888) μg/L vs (11.430 ±0.644) μg/L respectively,P ＜0.0l].At 4 and 24 hours,plasma levels of ACTH [(174.776-± 5.040) ng/L,(189.613 ± 4.802) ng/L respectively] in geneknock-out mice showed a higher increase than those in wild-type mice [(119.594 ± 6.945) ng/L,(124.93-± 11.001 7) ng/L respectively,P ＜ 0.05].Moreover,plasma levels of corticosterone [(40.138 ±-0.805) μg/L] at 4 hours and [(37.440-0.485)μg/L] at 24 hours in gene knock-out mice showed a same result as compared with that in wild-type mice [(19.702 ± 0.804) μg/L,(17.602 ± 0.743) μg/L respectively,P ＜ 0.05].Conclusions Knock-out of adenosine A2A receptor increases the release of ACTH and corticosterone in acute stage of moderate craniocerebral trauma and promotes pituitary-adrenal stress response.This may provide a novel explanation for the neuroprotective effect of A2A receptor deficiency.

BACKGROUND:The decel ularized porcine smal intestinal submucosa is a kind of bioactive extracel ular matrix, which is mainly composed of col agen, glycoprotein, proteoglycan and rich in col agen, glycosaminoglycan and various growth factors, and these components play an important role in promoting the differentiation and proliferation of tissue cel s. OBJECTIVE:To prepare the injectable smal intestinal submucosa and to investigate its co-culture with rat adipose-derived mesenchymal stem cel s in vitro. METHODS:The injectable smal intestinal submucosa and rat adipose-derived stem cel s were prepared. Cel counting kit-8 test for cel proliferation:Passage 3 adipose-derived stem cel s were seeded onto the injectable smal intestinal submucosa (experimental group) and cel s cultured under normal condition as control group. The cel proliferation was observed at 1, 3, 5 and 7 days of incubation. Live/dead staining test for the survival of cel s:Passage 3 adipose-derived stem cel s were respectively cultured in the injectable smal intestinal submucosa extracts (experimental group) and complete culture medium (control group). Cel survival was determined at 1, 3, 5 and 7 days of culture. RESULTS AND CONCLUSION:Scanning electron microscope oval and strip adipose-derived stem cel s adhered onto the material. The absorbance values in the experimental group were higher than those in the control group at 1 and 5 days of incubation (P<0.05). Cel survival:The number of cel s appeared to be in a rising trend with time in both two groups;after 1-day co-culture, al cel s in the two groups survived. Then dead cel s appeared in both two groups, showing no significant difference. These results show that the injectable smal intestinal submucosa exhibits a good cytocompatibility.

Aim To explore the protective effect of lo-ganin ( an active component in Cornus officinalis ) on podocyte injury induced by advanced glycation end products ( AGEs) and its possible mechanism. Meth-ods Mouse podocytes were cultured in vitro and di-vided into Normal group, model group ( AGEs group) , loganin group and aminoguanidine group ( set as posi-tive control) . After being incubated with loganin( final concentrations are 0. 1, 1, 10 μmol · L-1 ) for 1 h, podocytes were stimulated by AGEs of 100 mg · L-1 for 24 h. Then, the cell viability was measured by u-sing MTT method. Podocytes apoptosis was evaluated by Hoechst33342/PI staining and flow cytometry. Re-ceptors of advanced glycation end products ( RAGE ) ,desmin and apoptosis-related protein like Bax, Bcl-2, cleaved caspase-3 in podocytes were detected by Western blot. Results Loganin ameliorated podocyte injury induced by AGEs, down-regulated the expression of desmin and RAGE. Loganin also reduced the apoptotic rate of podocytes and decreased the ratio of Bax/ Bcl-2 and the expression of pro-apoptotic protein cleaved caspase-3 in podocytes. Conclusion Loganin could ameliorate podocyte injury, and its mechanism may be related to the decrease of the expression of RAGE and inhibition of the apoptotic pathway.

A colistin producing strain Paenibacillus polymyxa AS1.541 was treated by N-methyl-N-nitro-N-nitrosoguanidine(NTG) for increasing yields of the antibiotic colistin.High-yield strains were obtained by selection of deregulated mutant which grow on media containing colistin,a self second metabolite,and ethionine,an analogue of methionine.Some of these mutants have higher yield of colistin than that of the parent strain.

Objective: To establish a prediction model for high-risk cardiovascular disease (CVD) among residents aged 35 to 75 years, so as to provide the basis for improving CVD prevention and control measures. Methods: Permanent residents aged 35 to 75 years were selected from Dongcheng District, Beijing Municipality using the stratified random sampling method from 2018 to 2023. Demographic information, lifestyle, waist circumference and blood biochemical indicators were collected through questionnaire surveys, physical examinations and laboratory tests. Influencing factors for high-risk CVD among residents aged 35 to 75 years were identified using a multivariable logistic regression model, and a prediction model for high-risk CVD was established. The predictive effect was evaluated using the receiver operating characteristic (ROC) curve. Results: A total of 6 968 individuals were surveyed, including 2 821 males (40.49%) and 4 147 females (59.51%), and had a mean age of (59.92±9.33) years. There were 1 155 high-risk CVD population, with a detection rate of 16.58%. Multivariable logistic regression analysis showed that gender, age, smoking, central obesity, systolic blood pressure, fasting blood glucose, triglyceride and low-density lipoprotein cholesterol were influencing factors for high-risk CVD among residents aged 35 to 75 years (all P<0.05). The area under the ROC curve of the established prediction model was 0.849 (95%CI: 0.834-0.863), with a sensitivity of 0.693 and a specificity of 0.863, indicating good discrimination. Conclusion The model constructed by eight factors including demographic characteristics, lifestyle and blood biochemical indicators has good predictive value for high-risk CVD among residents aged 35 to 75 years.

OBJECTIVE: To explore the relationship between best corrected visual acuity (BCVA) and refraction parameters in myopia. METHODS: Two thousand two hundred and seventy-four patients (4245 eyes) with different degrees of myopia were collected. Their BCVA, diopter of spherical (DS), diopter of cylinder (DC), astigmatism axis, axial length (AL) and corneal thickness were detected. The influence of those parameters on BCVA was studied and the mathematical model of the relationship between BCVA and other parameters including the age and gender of patients was established. RESULTS: The logistic regression analysis showed that there were correlations between the BCVA (y) and DS (x1), DC (x2), gender (x3), AL (x4), corneal thickness (x5), astigmatism axis (x6) and age (x7) (P<0.05): y=0.580 6-0.034 0 x1-0.046 8 x2+0.056 5 x3+0.016 5 x4+ 0.0007 x5+0.000 2 x6-0.005 8 x7. CONCLUSION For people with myopia, age, gender and corneal thickness have small effect on BCVA, while the DS, DC, AL and astigmatism axis have significant effect on BCVA. The BCVA would decline following the extension of DS, DC and AL. It is helpful to assess the vision of myopia by analyzing the refraction parameters comprehensively.
Adolescent ; Adult ; Child ; Cornea/pathology* ; Female ; Forensic Medicine/methods* ; Humans ; Male ; Middle Aged ; Models, Theoretical ; Myopia/physiopathology* ; Refraction, Ocular/physiology* ; Refractometry ; Visual Acuity ; Visual Fields/physiology* ; Young Adult

Objective To observe the clinical effect of Tongyuan Acupuncture combined with Modified Linggui Zhugan Decoction in the treatment of polycystic ovary syndrome(PCOS)with spleen deficiency and phlegm-damp syndrome.Methods A total of 86 patients with PCOS of spleen deficiency and phlegm-damp type were randomly divided into observation group and control group,43 cases in each group.The control group was treated with Metformin Hydrochloride Tablets combined with Ethinylestradiol and Cyproterone Acetate Tablets.The observation group was treated with Tongyuan Acupuncture combined with Modified Linggui Zhugan Decoction on the basis of the treatment of the control group.Both groups were treated for three consecutive months.After three months of treatment,the clinical efficacy of the two groups was evaluated.The changes of sex hormone indexes estradiol(E2),testosterone(T),luteinizing hormone(LH),follicle stimulating hormone(FSH)and prolactin(PRL)expression levels were observed before and after treatment in the two groups.The changes of glucose metabolism levels including insulin resistance index(HOMA-IR),fasting insulin(FINS)and fasting blood glucose(FPG)expression levels were compared before and after treatment between the two groups.Results(1)The total effective rate was 90.70%(39/43)in the observation group and 74.42%(32/43)in the control group.The curative effect of the observation group was superior to that of the control group,and the difference was statistically significant(P＜0.05).(2)After treatment,HOMA-IR,FINS and FPG in the two groups were significantly improved(P＜0.05),and the improvement of HOMA-IR,FINS and FPG in the observation group was superior to that in the control group,the difference being significant(P＜0.05).(3)After treatment,the levels of E2,T,LH,FSH and PRL in the two groups were significantly improved(P＜0.05),and the levels of E2,T,LH,FSH and PRL in the observation group were significantly superior to those in the control group,the differences were statistically significant(P＜0.05).Conclusion Tongyuan Acupuncture combined with Modified Linggui Zhugan Decoction in the treatment of spleen deficiency and phlegm-damp type of PCOS can effectively improve the sex hormone level of patients and regulate the glucose metabolism of patients.The clinical effect is significant.

To improve the reliability and credibility of genotyping hepatitis E virus (HEV) and to explore the possibility of unifying standards of HEV genotyping by designing HEV universal primers for amplification of a long genomic fragment of different HEV genotypes. A set of universal primers (HEVuPrimer) was designed based on conserved regions determined by alignment analysis of 82 HEV strains with complete genome in GenBank. HEVuPrimer was compared with a set of previously used primers (MXJ primers) for their sequence-matching to different HEV strains and applied to amplify HEV genomic fragments from HEV reference strains with known different genotypes and clinical serum samples with anti-HEV-IgM by RT-nPCR. HEV genotyping based on the fragments amplified with HEVuPrimer was compared and validated with that based on HEV full genome and fragments obtained with MXJ primers. HEV genotyping by the phylogenetic analysis supplemented with the percent of nucleotide identity of the HEVuPrimer-determined fragments showed good correspondence with that based on HEV full-length genome. In addition, HEVuPrimer was much better than MXJ primers in matching sequences of HEV strains available from GenBank, and was able to amplify all the reference HEV strains with different genotypes. Among 124 samples with anti-HEV-IgM, 60 were positive for HEV RNA determined by a 644bp amplicon of RT-nPCR with the HEVuPrimenr. All the positive isolates belonged to HEV genotype 4 with nucleotide homology of 80.0%-99.9%, and could be further divided into 4 subgenotypes. Moreover, a novel subtype was identified with 6 HEV strains isolated very recently. The RT-nPCR using the HEVuPrimer and phylogenetic analysis of the amplified region provided strong evidences for its feasibility in HEV genetic classification. Our data have new implication for the consensus of genotype classification of HEV.
DNA Primers ; genetics ; Genome, Viral ; genetics ; Genotype ; Hepatitis E virus ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods

OBJECTIVETo observe effects of tetrandrine (Tet) on angiotensin II (Ang II)-induced cardiomyocyte hypertrophy and the activity and expression of phosphorylated ERK1/2 (p-ERK1/2).

METHODIn the primary culture of neonatal rat cardiomyocytes, as indexes of cardiomyocyte hypertrophy, pulsation rate was measured under phase contrast microscope. Cell size was determined by cell morphology analytical system. The total protein was determined by coomassie brilliant blue and protein synthesis rate was measured by [3H]-Leucine incorporation. ERK activity was measured by immuno-precipitation. The expression of p-ERK1/2 was assessed using Western blot.

RESULTTet can decrease Ang II-induced elevations of the pulsation rate, cell size, total protein and protein synthesis rate; inhibit the activity and expression of p-ERK1/2.

CONCLUSIONThe anti-hypertrophic effect of Tet on Ang II-induced cardiomyocyte hypertrophy was associated with inhibition of ERK1/2 signaling pathway.

Angiotensin II ; toxicity ; Animals ; Animals, Newborn ; Benzylisoquinolines ; isolation & purification ; pharmacology ; Blotting, Western ; Cell Size ; drug effects ; Cells, Cultured ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Hypertrophy ; Immunoprecipitation ; Microscopy, Phase-Contrast ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Phosphorylation ; drug effects ; Plants, Medicinal ; chemistry ; Protein Biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Stephania tetrandra ; chemistry