OBJECTIVE: The association between neutrophil-to-lymphocyte ratio (NLR) with subclinical macrovascular and microvascular diseases has been less investigated. We sought to examine the association between NLR and new-onset subclinical macrovascular and microvascular abnormalities in the Chinese population. METHODS: From a community cohort, we included 6,430 adults aged ≥ 40 years without subclinical macrovascular and microvascular diseases at baseline. We measured subclinical macrovascular and microvascular abnormalities separately using the ankle-brachial index (ABI), brachial-ankle pulse wave velocity (baPWV), and albuminuria. RESULTS: During a mean follow-up of 4.3 years, 110 participants developed incident abnormal ABI, 746 participants developed incident elevated baPWV, and 503 participants developed incident albuminuria. Poisson regression analysis indicated that NLR was significantly associated with an increased risk of new-onset abnormal ABI, elevated baPWV, and albuminuria. Compared to overweight/obese participants, we found a much stronger association between NLR and subclinical vascular abnormalities in participants with normal weight. Furthermore, we found an interaction between the NLR and body mass index (BMI) on the risk of new-onset abnormal ABI ( P for interaction: 0.01). CONCLUSION NLR was associated with subclinical macrovascular and microvascular diseases in the Chinese population. Furthermore, in participants with normal weight, the association between NLR and subclinical vascular abnormalities was much stronger.
Adult ; Aged ; Ankle Brachial Index ; Body Mass Index ; China/epidemiology* ; Cohort Studies ; Female ; Humans ; Incidence ; Lymphocytes/cytology* ; Male ; Middle Aged ; Neutrophils/cytology* ; Poisson Distribution ; Prospective Studies ; Vascular Diseases/etiology*

Objective To investigate the seroprevalence of Toxoplasma gondii infections among neonates in Fujian Province, so as to provide insights into the development of interventions for the prevention and control of congenital toxoplasmosis. Methods A total of 1 045 neonates delivered in Fujian Province from 2017 to 2018 were recruited, including 387 preterm infants and 658 full-term infants. Umbilical cord blood was sampled from all neonates, and the seroprevalence of anti-T. gondii IgG antibody was detected and compared between preterm and full-term infants. In addition, elbow venous blood samples were collected from neonates’mothers, and the seroprevalence of anti-T. gondii IgG antibody was detected and compared between preterm and full-term infants’mothers. Results The overall seroprevalence of anti-T. gondii IgG antibody was 9.38% among the 1 045 neonates in Fujian Province. The seroprevalence of anti-T. gondii IgG antibody was 18.35% in the 387 preterm infants, and there was no significant difference in the seroprevalence of anti-T. gondii IgG antibody between male and female infants (17.69% vs. 18.75%, χ² = 0.07, P > 0.05). The seroprevalence of anti-T. gondii IgG antibody was 4.10% in the 658 full-term infants, and there was no significant difference in the seroprevalence of anti-T. gondii IgG antibody between male and female infants (4.14% vs. 4.08%, χ² = 0, P > 0.05). In addition, the overall seroprevalence of anti-T. gondii IgG antibody was 15.02% in all neonates’ mothers, and the seroprevalence was significantly greater in preterm infants’mothers than in full-term infant’s mothers (20.93% vs. 11.55%, χ² = 16.79, P < 0.01). Conclusions The seroprevalence of T. gondii infections is significantly higher in preterm infants and their mothers than in full-term infants and their mothers. Prenatal detection of T. gondii infections and health education pertaining to toxoplasmosis prevention and control knowledge are required to be strengthened to effectively reduce the incidence of congenital toxoplasmosis.

Objective To prepare HDAg with biological activities as a candidate of diagnostic reagent.Methods To synthesize HDV gene fragment after codon optimization.To construct a thio-fused recombinant plasmid based on M48 expression vector.To express in E.coli induced by IPTG.To purify the protein by affinity chromatography followed by characterization in ELISA.Results Plasmid construction was verified by enzyme digestion.SDS-PAGE indicated the molecular weight of the protein was the same as we expectation.ELISA proved its affinity with HDV antibodies.Conclusion HDAg was obtained successfully and it will pave the road to the research of HDV diagnostic reagent.

Objective To study the effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering.Methods Based on the colon preference of E.coli,the HDV small antigen original gene from GenBank was optimized.Both the original gene and the optimized gene expressed in prokaryotic cells,SDS-PAGE was made to analyze the protein expression yield and to decide which protein expression style was more proportion than the other.Furthermore,two antigens were purified by chromatography in order to compare the purity by SDS-PAGE and Image Lab software.Results SDSPAGE indicated that the molecular weight of target proteins from two groups were the same as we expected.Gene optimization resulted in the higher yield and it could make the product more soluble.After chromatography,the purity of target protein from optimized gene was up to 96.3％,obviously purer than that from original gene.Conclusion Gene optimization could increase the protein expression yield and solubility of genetic engineering HDV small antigen.In addition,the product from the optimized gene group was easier to be purified for diagnosis usage.

Objective To establish a novel improved loop-mediated isothermal amplification (LAMP) technique to detect hepatitis A virus (HAV).Methods A novel improved LAMP assay based on the addition of an acceleration primer was developed for hepatitis A virus nucleotide acid detection.Results Precision and reproducibility analysis proved its high stability and reliability.Comparison between the improved and conventional LAMP assays revealed that the former was more sensitive with a detection limit of 5 TCID5o/ml.The novel detection method displayed 100％ consistency with the TaqMan real-time PCR assay when applied to clinical specimens collected from patients with confirmed acute HAV infection.Conclusion This novel technique is widely applicable as a simple diagnostic tool in the clinical field as well as for the surveillance and investigation of the infectious disease in developing countries where HAV is endemic.

Objective To construct virus-like particles of hepatitis B core antigen, with HBsAg "a" epitope exposed on the surface. Methods Hepatitis B surface antigen "a" epitope were inserted into the Hepatitis B core antigen, between the 78th (Asp) and the 79th (Pro) amino acids. The gene was synthesized after the codon optimized, then it was ligated to the express vector after been enzyme digest. The virus-like particles were observed by electron microscope and detected by ELISA after been expressed and purified. Immune the rabbits by the VLPs, then detect the antibody. Result The virus-like particles were confirmed by electron microscope. Its antigenicity and immunogenicity were identified by ELISA. Conclusion The prokaryotic express plasmid with the fusion gene has been constructed successfully. The virus-like particles have been expressed, purified and identified, which lays the foundation for its application in the further.

Objective Obtain the hepatitis C virus high purified subunit fusion protein and detect its immunogenicity.Methods With the vector of pET-11d,fusion protein was expressed in Escherichia coli BL21(DE3) after induced by IPTG.The protein was then purified by DEAE negative ion exchange chromatography and Ni2+ affinity chromatography.Western Blot analysis was used to detect the antigenicity of the fusion protein.At the same time,the sera were collected and prepared from the immunized experimental animals in order to investigate the immunogenicity of the protein by EIA.Results High purified hepatitis C virus subunit fusion protein was obtained successfully.The EIA indicated that the fusion protein could elicit specific antibodies in the animals with very high titers.Conclusion The hepatitis C virus subunit fusion protein expressed in prokaryotic system was proved to have strong immunogenicity.It could provide some helpful and useful information to the hepatitis C virus prophylactic and therapeutic vaccine development.

Objective To prepare monoclonal antibodies (McAbs) against VP1 capsid protein of Enterovims 71. Methods Two peptides, SP55 and SPT0, containing amino acid 163-177 and 208-222 of VP1, were synthesized respectively. Immunized BALB/c mice with the synthetic peptides to estabhsh the hybridoma cell strains secreting specific McAb to VP1. After the specific McAbs were prepared, identified and analyzed the titer by indirect ELISA assay. The positive clones were selected and their neutralization titer were determined by neutralization test. Results Two high titered anti-VP1 antibodies secreted by the hybfidoma cells showed good neutralization reaction with enterovirus 71 on RD cells, and the neutralization titer were 1:8 and 1 : 16 respectively. Conclusion Two high titered anti-VP1 antibodies, with good neutralization activity, secreted by the hybridoma cells, which lays the foundation for further study.

OBJECTIVETo study the antigenic properties of mutant hepatitis B virus surface antigen, to understand the sensitivity of the commercially available HBsAg assays to the variants and to reduce the undetectability of the variants.

METHODSRecombinant eukaryotic expression plasmids for HBsAg. The recombinant eukaryotic expression plasmids pSS1adr, pSS1adw2, pSS1adw2- 145Arg, pSS1adr-126 Asn and pSS1adr-126Ser were transfected into COS-7 cells. HBsAg in the supernatants of transfected cells was detected by using different commercial ELISA kits.

RESULTSThe absorbance value of pSS1adr-126 Asn and pSS1adr-126Ser plasmids were similar to that of the wild type HBsAg, the absorbance value of pSS1adw2-145Arg plasmids was lower than that of the wild type HBsAg.

CONCLUSIONIt is estimated that the antigenicity of HBsAg mainly depended on the amino acid sequence of "a" antigen determinant and its conformation, so 145 amino acid substitutions led to the change of conformation and the antigenicity of variant HBsAg was lower than that of the wild type.

Animals ; COS Cells ; Cercopithecus aethiops ; Culture Media, Conditioned ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; genetics ; metabolism ; Hepatitis B Surface Antigens ; analysis ; genetics ; immunology ; Mutation ; Transfection ; Viral Envelope Proteins ; genetics

OBJECTIVETo clone human PD-1 gene, construct a prokaryotic expression plasmid and express in E. coli.

METHODSThe human PD-1 cDNA was cloned by RT-PCR from the total RNA, which was extracted from peripheral blood lymphocyte cell of the patient with chronic hepatitis B. Recombinant PD-1 protein was been expressed and purified after the prokaryotic expression plasmid had been constructed. It was identified by SDS-PAGE, DNA sequencing and amino acid sequencing.

RESULTSThe PD-1 gene was cloned and confirmed by DNA sequencing. The recombinant protein was expressed in E. coli. The purified protein was obtained, then been confirmed by amino acid sequencing.

CONCLUSIONThe human PD-1 gene was successfully cloned and expressed in E. coli, which lays the foundation for further study on the function and application of PD-1.

Amino Acid Sequence ; Antigens, CD ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Apoptosis Regulatory Proteins ; biosynthesis ; chemistry ; genetics ; isolation & purification ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Genetic Vectors ; genetics ; metabolism ; Humans ; Polymerase Chain Reaction ; Programmed Cell Death 1 Receptor ; Prokaryotic Cells ; metabolism ; Sequence Alignment ; Sequence Analysis, DNA