Study on the New Clinical Significance of Routine Indexes in Pregnant Women with HDP before 20 Weeks of Pregnancy / 现代检验医学杂志

Objective To explore the new clinical significance of these routine test indexes and find the early biomarkers for predicting hypertension disorder complicating pregnancy(HDP) by the comparative analysis between pregnant women with HDP and normal pregnant women before 20 weeks,including biochemical,blood coagulation and blood routine indexes.Methods A case-control study was conducted to review the clinical data of pregnant women who were undertaken prenatal examination and delivered in Guangzhou Women and Children's Medical Center from 2012 to 2016.The pregnancy were divided into HDP group and control group according to the gestational week and age.Analyzed the related test indexes before 20 weeks of pregnancy.The two groups variables were analyzed by Kolmogorov-Smirnov for normal distribution.Approximate measurement data of normal distribution were mean± standard deviation (x± s) for statistical description,while the data of the skewed distribution was median and percentile[M(P25～P75)]for statistical description.The diagnostic value was analyzed by logistic regression equation and receiver operating characteristic curves (ROC).Results The level of biochemical indexes (AST,γ GT and UA) of HDP group was significantly higher thanthe normal group (t=2.50,3.34,4.56,P＜ 0.05).Meanwhile the level of blood indexes (RBC,PLT and HCT) of HDP group was significantly higher than the normal group (t=2.89,4.51,3.29,all P＜0.01).Other indicators of two groups (ALT,TBIL,Cr,Urea,PT,APTT,TT,FIB and HGB) were not significantly different (t=0.25～ 1.85,all P＞0.05).Then the logistic regression model equation was Y=-5.497+0.010 * PLT+0.043 * γ-GT+0.007 * UA+0.045 * AST.The area under ROC curve (AUC) was 0.746 for the combination of the four indexes.The combination resulted in a higher sensitivity of 0.818 and specificity of 0.523.Conclusion Before 20 weeks of gestation,compared with normal group,liver and kidney function in patients with HDP and high blood coagulation state was damaged.Can by combined detection of AST,UA,γ-GT and PLT index,early prevention and diagnosis of gestational hypertension,effective intervention measures taken as soon as possible to improve the prognosis of pregnancy.

Two novel antimicrobial peptides from skin venoms of spadefoot toad Megophrys minor / 中国天然药物

Amphibian skin contains rich bioactive peptides. Especially, a large amount of antimicrobial peptides have been identified from amphibian skin secretions. Antimicrobial peptides display potent cytolytic activities against a range of pathogenic bacteria and fungi and play important defense roles. No antimicrobial peptides have been reported from toads belonging to the family of Pelobatidae. In this work, two novel antimicrobial peptides (Megin 1 and Megin 2) were purified and characterized from the skin venoms of spadefoot toad Megophrys minor (Pelobatidae, Anura, Amphibia). Megin 1 had an amino acid sequence of FLKGCWTKWYSLKPKCPF-NH2, which was composed of 18 amino acid residues and contained an intra-molecular disulfide bridge and an amidated C-terminus. Megin 2 had an amino acid sequence of FFVLKFLLKWAGKVGLEHLACKFKNWC, which was composed of 27 amino acid residues and contained an intra-molecular disulfide bridge. Both Megin 1 and Megin 2 showed potential antimicrobial abilities against bacteria and fungi. The MICs of Megin 1 against Escherichia coli, Bacillus dysenteriae, Staphylococcus aureus, Bacillus subtilis, and Candida albicans were 25, 3, 6.25, 3, and 50 μg·mL(-1), respectively. The corresponding MICs for Megin 2 were 6.25, 1.5, 12.5, 1.5, and 12.5 μg·mL(-1), respectively. They also exerted strong hemolytic activity against human and rabbit red cells. The results suggested that megin peptides in the toad skin of M. minor displayed toxic effects on both eukaryotes and prokaryotes. This was the first report of antimicrobial peptides from amphibians belonging to the family of Pelobatidae.

Effect of poly(DL-lactide-co-glycolide) on scar formation after glaucoma filtration surgery / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Glaucoma filtering surgery (GFS) is the most common procedure performed in the treatment of glaucoma. Although antiscarring agents help prevent postsurgical scarring and improve glaucoma surgical outcomes, they may be associated with an increased incidence of severe and potentially blinding complications. Poly(DL-lactide-co-glycolide) (PDLLA/GA) is a bioresorbable polymer, which can be prepared with a large range of physical, mechanical, and biological properties and has been widely used in medicine, including as an absorbable suture and a drug carrier and especially as a scaffold in tissue engineering. This study aimed to evaluate the effect of PDLLA/GA on scar formation after glaucoma filtration surgery (GFS).</p><p><b>METHODS</b>Forty-eight New Zealand white rabbits were divided into two groups randomly and GFS was performed on the right eye of each. PDLLA/GA membranes were put under the sclera flap for evaluation. GFS with no membrane inserted served as control. Clinical evaluations of intraocular pressure (IOP) and the presence of a filtration bleb were performed at intervals (3 days, 1, 2, 4, 8, 12, 20, and 24 weeks) postoperatively. At each time point, three eyes per group were excised to observe histological changes such as inflammation and scar formation and the expression of collagen type IV, proliferating cell nuclear antigen (PCNA), matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-1). The expression of connective tissue growth factor (CTGF) mRNA was determined by reverse transcription-polymerase chain reaction.</p><p><b>RESULTS</b>The lower IOP level and an effective bleb were maintained for a long time after GFS in the PDLLA/GA group. The histological analysis showed less inflammation and scar formation, weaker expression of collagen type IV and PCNA, more intense MMP-9 and TIMP-1, slightly elevated ratio of MMP-9 and TIMP-1, and a smaller increase in CTGF mRNA postoperatively in the PDLLA/GA group but less than the control group (P < 0.05).</p><p><b>CONCLUSION</b>PDLLA/GA membranes may be promising for preventing fibrosis after GFS.</p>

Correlation of serum IL-18 level and IL-18 gene promoter polymorphisms to the risk of cervical cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the correlations of serum interleukin-18 (IL-18) level and IL-18 gene promoter polymorphisms to the development of cervical cancer (CC).</p><p><b>METHODS</b>Five single nucleotide polymorphisms (SNPs) of the IL-18 gene promoter region at position rs5744224, rs1946519, rs1946518, rs5744225 and rs5744226 were detected by means of sequences analysis in 50 CC patients and 50 normal subjects that matched for age and residence, and their serum IL-18 level was tested using enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Two linkage SNP sites (rs1946519 and rs1946518) in the up-stream of IL-18 gene were identified to present 3 genotypes, namely TT-AA, GG-CC, and TG-AC. A significant difference in the frequency of the 3 genotypes was observed between the CC patients and the normal controls (chi 2=17.497, P=0.000). The frequency of T (A)/G (C) alleles was 42% (42/100) in normal controls and 73% (73/100) in patients, showing significant diference (chi 2=19.662, P=0.000). Serum IL-18 concentrations of the CC patients was significantly lower than that of the normal controls (95.470-/+18.827 vs 116.756-/+16.262 pg/ml, F=14.445, P=0.000). In the cancer patients, serum IL-18 was 90.668-/+20.363 pg/ml for TT-AA genotype, 119.641-/+15.338 pg/ml for GG-CC genotype, and 112.793-/+13.326 pg/ml for TG-AC genotype, showing significant differences (F=11.307, P=0.000). A significant interaction effect was suggested between IL-18 genotype and CC (F=4.223, P=0.018).</p><p><b>CONCLUSION</b>IL-18 gene polymorphisms and serum IL-18 level are related to the development of CC, and two SNPs, rs1946518 and rs1946519, can be important genetic factors for the susceptibility of cervical neoplasms.</p>

Relationship between surface molecules and RelB gene expression in DC2.4 cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the relationship between the expression of the nuclear factor-kappaB transcription factor RelB gene and the surface molecules in DC2.4 cell line.</p><p><b>METHODS</b>DC2.4 cell line was cultured in complete RPMI 1640 medium, whose morphology was observed with optical microscope and the intracellular structures with transmission electron microscope. Flow cytometry was performed to analyze the surface markers of the cells, including MHC-II, CD86 and CD40, and RelB mRNA expression was detected by RT-PCR.</p><p><b>RESULTS</b>Under optical microscope, the cells appeared irregular in shape with obvious dendritic cell processes, and electron microscopy revealed homogenous fat drops and phagocytic vesicles in the cytoplasm. Flow cytometry demonstrated low expression levels of MHC-II and CD40, but high level of CD86 molecules. Low-level expression of RelB mRNA was detected by RT-PCR, resembling its expression level in bone marrow-derived DC with immature phenotype.</p><p><b>CONCLUSION</b>DC2.4 is a mouse bone marrow dendritic cell line with strong phagocytic capacity, and the low expression of both RelB gene and surface CD40 molecules suggests an immature dendritic cell line.</p>

Association of interleukin-18 gene rs1946519 and rs360718 single nucleotide polymorphism with cervical cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the association of interleukin-18 (IL-18) rs1946519 and rs360718 single nucleotide polymorphisms (SNPs) with cervical cancer and the risk factors for cervical cancer.</p><p><b>METHODS</b>The genotypes of IL-18 gene SNPs were detected in 107 cervical cancer patients and 80 healthy female blood donors (control group) by TaqMan and MGB probe assays. The allele and genotype frequencies and risk factors of cervical cancer were analyzed by Chi-square test in both groups.</p><p><b>RESULTS</b>No significant differences were found in rs360718 allele and genotype frequencies between patients with cervical cancer and the control group (P>0.05), but the allele and genotype frequencies in rs1946519 as well as the genotypes of rs1946519 and rs360718 (AC+TT) were found to relate to cervical cancer (P<0.05).</p><p><b>CONCLUSION</b>rs360718 allelic variation in IL-18 gene does not contribute to the susceptibility of cervical cancer in these patients, but rs1946519 allelic variation can be closely related to the pathogenesis of cervical cancer. Women with AC+TT genotype are at relatively high risk for cervical cancer.</p>

Construction and identification of RNA interference vector for human tissue factor gene / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a RNA interference vector for human tissue factor (TF) gene.</p><p><b>METHODS</b>Human TF short hairpin RNA (shRNA) sequence was designed using online design software (Invitrogen) and synthesized into double-strand oligonucleotide (ds oligo), which was cloned into the pENTRTM/U6 plasmid, followed by transformation of the product into competent Top10 E. coli cells. After expansion of the transformed bacteria, the plasmid was extracted and sequenced, which was subsequently transfected into human umbilical vein endothelial cells (HUVECs). The interference effect of the vector on the target gene expression was detected by RT-PCR and immunofluorescence assay.</p><p><b>RESULTS</b>The sequencing result indicated that the plasmid pENTRTM/U6-RelB-shRNA was constructed correctly, which resulted in effective inhibition of TF expression in HUVECs after transfection.</p><p><b>CONCLUSION</b>The RNA interference vector against human TF gene has been constructed successfully, which may provide a stable transfection vector for potential treatment of blood coagulation abnormalities.</p>

Identification of effective siRNA sequence for RelB silencing in murine dendritic cells with siRNA cassette / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct small interfering RNA (siRNA) expression cassette targeting murine RelB gene and identify the most effective siRNA sequence against RelB gene in murine bone marrow-derived dendritic cells (DCs).</p><p><b>METHODS</b>Three expression cassettes namely R1/siRNA, R2/siRNA and R3/siRNA targeting the sites 1027, 302 and 1121 of RelB gene, respectively, were constructed by PCR approach and transfected into cultured murine myeloid DCs by catione liposome Advant-Gene. After incubation for 24 hours in a incubator containing 5% CO(2) at 37 degrees C, the DCs were stimulated by lipopolysaccharide (LPS), and RelB gene expression in DCs were then detected by RT-PCR and immunofluorescence.</p><p><b>RESULTS</b>RT-PCR and immunofluorescence assay showed that the expression of RelB gene in DCs transfected with R2/siRNA could not be upregulated by LPS stimulation, but transfection with R1/siRNA or R3/siRNA failed to produce such effect.</p><p><b>CONCLUSION</b>R2/siRNA is an effective sequence for RelB silencing, and can be a useful means to construct new tolerogenic DC, RNAi RelB DC, for clinical immunotolerance induction.</p>