Objective To investigate the correlation between the single nucleotide polymorphisms (SNPs) of IL-4 gene -590C/T as well as IL-13 gene -1112C/T and the incidence of asthma in children.To address whether the SNPs of two loci have any impact on total serum IgE( TIgE) levels.Methods Polymerase chain reaction-restriction fragment length polymorphism technique( PCR-RFLP) was used to determine the two locus polymorphisms of 250 patients with asthma and 200 healthy subjects in control group.The level of TIgE was tested by enzyme-linked immunosorbent assay(ELISA) in two groups.Results (1)The genotype distribu-tion in each locus was different in two groups(P<0.01).(2)In patient and control groups, the C allele fre-quency at IL-4-590 locus was 23.00%and 35.25%,the T allele frequency at the same locus was 77.00%and 64.75%, respectively.Besides, the C allele frequency at IL-13 -1112 locus was 59.20%and 70.50%,the T allele frequency at the same locus was 40.80%and 29.50%, respectively.The allele frequencies at each locus had the significant difference in asthma and control group(P<0.01).(3)The subjects with variant allele have higher risks for asthma than those without variant allele ( OR=1.82, 95%CI=1.36-2.44, and OR=1.647, 95%CI=1.246-2.178).(4)The TIgE level of asthma group was higher than that of control group in those with the same genotype (P<0.05).(5)No significant difference with the TIgE levels was observed in those with or without variant T allele at IL-13 -1112 locus(P>0.05), which was in contrast with that at IL-4 -590 locus in asthma group(P<0.05).Conclusion The SNPs of two loci were associated with childhood asthma.Variant al-lele T at 590C/T locus in IL-4 gene correlated with higher serum IgE levels.There was no significant correlation between the serum IgE levels and variant T allele at IL-13 -1112C/T locus.

Objective To evaluate the relationship between varicocele (VC) and the prostativenouplexuby coloDoppleflow imaging(CDFI) and to explore the etiology of varicocele .MethodThe innediameterand the hemorrheologiparameterof spermativein and prostativenouplexuwere observed in 135 patientwith lefvaricocele(lefVgroup) ,51 patientwith bilat-eral V(bilateral Vgroup) and the control group(100 cases) by CDFI .The diameteof the prostativenouplexus(PVD) ,peak velocity of reflux flow (RFV) in the Valsalvtesand the peak velocity of antegrade flow (AFV) aresin 3 groupwere statistical-ly analyzed .ResultPVD and RFV in the bilateral Vgroup were greatethan those in the lefVgroup and the control group (P＜0 .01) .PVD and RFV in the lefVgroup had no statistical differencecompared with the control group (P＞0 .05) .AFV had no statistical difference among 3 group(P>0 .05) .PVD ,RFV and AFV in 30 caseof Vhad no statistical differencebe-tween before and afteoperation (P>0 .05) .Conclusion Bilateral Vmay be accompanied with potential systematic vascular abnormalities.

Objective To high express and purify toxoplasma gondii antigen GRA6 in E.coli which can be used to develop the genetic engineering diagnostic reagents.Methods The recombinant plasmid of pGEX-GRA6 was transformed to a bacterium BL21-Codon Plus(DE3)-RP and the recom- binant product was expressed under the inducement of isopropyl-beta-D-thiogalactosidase(IPTG). Cells were lysed by multiple rounds of sonication.The expression product was analyzed by using SDS- PAGE.Furthermore,it was purified by sedimentation of ammonium sulphate,desalting using Sephe- dax GS0 and affinity chromatography on glutathione-sepharose system.The immunogenicity of recom binant antigen purified was tested with Western blot.Results GRA6 was highly expressed in E.coli as fusion protein consisting of glutathione S-transferase and GRA6(GST-GRA6).The solubility anal- ysis of expression product indicated that this recombinant protein could be expressed in both superna- tant and inclusion bodies.The soluble protein of GRA6 in supernatant yield the final preparation at greater than 90% after purification.The recombinant protein purified could be recognized by human toxoplasmosis-infective sera with Western blotting analysis.Conclusions The soluble protein of GRA6 was highly expressed in E.coli and the recombinant antigen could be recognized by human tox- oplasmosis-infective serum after purification.The recombinant antigen can be used for devoloping kit to diagnose the acute and chronic infection of toxoplasma gondii.

Objective To explore ultrasonic image for the normal anatomy of the larynx,and provide the basis of ultrasonic diagnosis in laryngeal diseases.Methods Ultrasound anatomy for the larynx was established by way of comparing the structures of four corpses and ultrasonic imaging of the larynx of normal control group.Results Ultrasonic image for the normal anatomy of the larynx was established by comparing the anatomy tomography of corpses and ultrasonic imaging of the larynx of normal control group.Conclusions Ultrasonography could be applied in the examination of the laryngeal diseases as it could show unambiguous ultrasonic imagings of the larynx,and adding an important complementary technique to clinical medicine.

Objective To prepare a nano drug(PFOB@Lip-MMC)with liposome as the carrier,liquid perfluorooc-tyl bromide(PFOB)as core and mitomycin C(MMC)loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts(HPFs).Methods The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@Lip-MMC and detect its physical and chemical properties.Cell Counting Kit-8,Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@Lip-MMC on the via-bility of HPFs.DiI fluorescence labeled PFOB@Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope.After establishing HPF inflammatory cell models,they were divided into the control group(with sterile phosphate-buffered saline solution added),PFOB@Lip group(with PFOB@Lip added),MMC group(with MMC added),PFOB@Lip-MMC group(with PFOB@Lip-MMC added)and normal group(with fresh culture medi-um added)according to the experimental requirements.After co-incubation for 24 h,flow cytometer was used to detect the apoptosis rate of inflammatory cells,and the gene expression levels of interleukin(IL)-1β,prostaglandin E2(PGE2),tumor necrosis factor(TNF)-α and vascular endothelial growth factor(VEGF)in cells were analyzed by PCR.Results The average particle size and Zeta potential of PFOB@Lip-MMC were(103.45±2.17)nm and(27.34±1.03)mV,respec-tively,and its entrapped efficiency and drug loading rate were(72.85±3.28)％and(34.27±2.04)％,respectively.The sustained-release MMC of drug-loaded nanospheres reached(78.34±2.92)％in vitro in a 24-hour ocular surface environ-ment.The biological safety of PFOB@Lip-MMC significantly improved compared to MMC.In terms of the DiI fluorescence labeled PFOB@Lip-MMC,after co-incubation with inflammatory HPFs for 2 h,DiI fluorescence labeling was diffusely dis-tributed in the cytoplasm of inflammatory HPFs.The apoptosis rate of inflammatory HPFs in the PFOB@Lip-MMC group[(77.23±4.93)％]was significantly higher than that in the MMC group[(51.62±3.28)％].The PCR examination results showed that the gene transcription levels of IL-1 β,PGE2,TNF-α and VEGF in other groups were significantly reduced com-pared to the control group and PFOB@Lip group,with the most significant decrease in the PFOB@Lip-MMC group(all P＜0.05).Conclusion In this study,a novel nano drug(PFOB@LIP-MMC)that inhibited the proliferation of HPFs was successfully synthesized,and its cytotoxicity was significantly reduced compared to the original drugs.It has good bio-compatibility and anti-inflammatory effects,providing a new treatment approach for reducing the recurrence rate after pte-rygium surgery.

[Objective] To determine the feasibility of preparing plasminogen (Pg) with Fraction Ⅲ precipitation (hereinafter referred to as FⅢ-P) from low-temperature ethanol process by full chromatography (hereinafter referred to as FⅢ-P process). [Methods] The FⅢ-P was diluted with dissolution buffer at different dilution times and stirring time. The potency and antigen concentration of Pg in dissolution sample were detected and the dissolution and clarification conditions were determined. Pre-treatment of loading sample and pre-experiment of affinity chromatography were carried out on the FⅢ-P dissolution sample to judge whether the loading sample had an impact on the chromatography by observing the performance of the affinity chromatography column and to evaluate whether the affinity chromatography could achieve the purpose of purifying Pg by detecting the plasma protein antigen concentration and Pg potency of the samples in the process. Two batches of FⅢ-P process were studied step by step, and the specific activity, steps and total recovery, and the output of Pg per ton of plasma were calculated. The feasibility of preparing Pg by FⅢ-P process was evaluated by comparing with the data of full chromatography process using plasma as raw material (hereinafter referred to as plasma process). [Results] The FⅢ-P was dissolved with 10 times of dissolution buffer, stirred for 1 hour, centrifuged at room temperature of 10 000×g for 15 minutes. The supernatant was first filtered with a screen, then clarified with an 8/0.8 μm filter, and finally filtered with a 0.45/0.2 μm filter and loaded. Pre-test showed that from clarification and filtration to Pg affinity chromatography, the step recovery of activity and antigen was 39.51% and 108.64%, respectively, the antigen concentration of Pg increased by 31.16 times and the activity increased by 11.39 times after affinity chromatography, which reaching the effect of affinity chromatography purification of Pg. The results of 2 batches of step-by-step scale-up FⅢ-P process showed that the total recoveries of antigen and activity from plasma to SP chromatography of FⅢ-P process were (45.76±1.10)% and (24.15±0.59)%, respectively, which had a total loss of about 1/3 of antigen and about 2/3 of activity compared to the plasma process. The Pg specific activity of SP chromatography eluent was (4.68±0.25) U/mg, which was about half of that of plasma process, but meeted the internal standard of > 4 U/mg. The output of Pg antigen per ton of plasma in the FⅢ-P process was 68.73% of that in the plasma process, and the output of Pg activity per ton of plasma in the plasma process was 29.82% of that in the plasma process, which basically achieved the purpose of waste utilization of FⅢ-P. [Conclusion] The technical route of preparing Pg from FⅢ-P by full chromatography is feasible.

【Objective】 To map a comprehensive description of microbiome presented in healthy blood donations in Liangshan Yi Autonomous Prefecture, and detect the potential pathogens and its prevalence. 【Methods】 A total of 1 299 blood samples were randomly selected from healthy blood donors in Liangshan Prefecture. Total nucleic acids were extracted and sequenced by high-throughput sequencing, and the microbiome was determined by metagenomics analysis. Quantitative real-time PCR (qPCR) and ELISA assays were used to detect antibodies against HSV DNA and HSA in each sample. The prevalence of HSV in healthy blood donors was compared in terms of gender, age, occupation, education level, and frequency of donation. 【Results】 3.49GB data were obtained from DNA pool through high-throughput sequencing. After filtering the data of human genome, the DNA sequences annotated as bacteria, parasites / fungi and viruses were 213 057, 10 623 and 15 reads, respectively. A total of 2.79GB data were obtained from cDNA pool, after filtering the data of human genome, the fragments annotated as bacteria, parasites / fungi and viruses were 4 105 600, 18 446 and 0 reads, respectively. The prevalence of IgG and / or IgM antibodies against herpes simplex virus type 1 and 2 were 8.62% (112 / 1 299) and 18.32% (238/1 299), and that of nucleic acid was 0.77 ‰ (1/1 299). 【Conclusion】 The microbiome of healthy blood donors in Liangshan Prefecture and the potential pathogens were identified in this study. Regional specificity of HSV infections emerged in Liangshan Prefecture. Considering the collaboration between HSV-2 and HIV infection, epidemiological investigation of HSV-2 infection should be conducted preferentially among different populations in Liangshan Prefecture and other HIV high prevalence areas in order to benefit the prevention and control of HIV.

Melatonin receptor 1B (MT2, encoded by the MTNR1B gene), a high-affinity receptor for melatonin, is associated with glucose homeostasis including glucose uptake and transport. The rs10830963 variant in the MTNR1B gene is linked to glucose metabolism disorders including gestational diabetes mellitus (GDM); however, the relationship between MT2-mediated melatonin signaling and a high birth weight of GDM infants from maternal glucose abnormality remains poorly understood. This article aims to investigate the relationship between rs10830963 variants and GDM development, as well as the effects of MT2 receptor on glucose uptake and transport in trophoblasts. TaqMan-MGB (minor groove binder) probe quantitative real-time polymerase chain reaction (qPCR) assays were used for rs10930963 genotyping. MT2 expression in the placenta of GDM and normal pregnant women was detected by immunofluorescence, western blot, and qPCR. The relationship between MT2 and glucose transporters (GLUTs) or peroxisome proliferator-activated receptor γ (PPARγ) was established by western blot, and glucose consumption of trophoblasts was measured by a glucose assay kit. The results showed that the genotype and allele frequencies of rs10830963 were significantly different between GDM and normal pregnant women (P<0.05). The fasting, 1-h and 2-h plasma glucose levels of G-allele carriers were significantly higher than those of C-allele carriers (P<0.05). Besides, the protein and messenger RNA (mRNA) expression of MT2 in the placenta of GDM was significantly higher than that of normal pregnant women (P<0.05). Melatonin could stimulate glucose uptake and GLUT4 and PPARγ protein expression in trophoblasts, which could be attenuated by MT2 receptor knockdown. In conclusion, the rs10830963 variant was associated with an increased risk of GDM. The MT2 receptor is essential for melatonin to raise glucose uptake and transport, which may be mediated by PPARγ.
Female ; Humans ; Pregnancy ; Blood Glucose/metabolism* ; Diabetes, Gestational/metabolism* ; Glucose/metabolism* ; Melatonin/metabolism* ; Polymorphism, Genetic ; PPAR gamma ; Receptor, Melatonin, MT2/genetics*