Two methods of different RNA extractions from Conus venom tube and toxin gland of seven cone snail species were performed. ThermoScript~ TM RNase H~ - Transcriptase and AMV Transcriptase were used to synthesize cDNA separately. The cDNAs were used as templates to amplify conotoxin genes by PCR.The optimized RNA isolation and cDNA synthesis methods were obtained according to RT-PCR results, which would be the basis for cloning of new conotoxin genes.

Objective:To analyze the report accuracy and risk factors of congenital syphilis (CS), and to provide references of accurate diagnosis for clinicians.Methods:CS notification data, including suspected cases and confirmed cases, were obtained from national notifiable disease reporting system (NNDRS) reported from 2016 to 2019 in Guangzhou City. Information of syphilis infected pregnant women and their children who were diagnosed or excluded with CS during 2012-2019 were extracted from Guangzhou City'the integrated prevention of mother-to-child transmission of human immunodeficiency virus, syphilis and hepatitis B virus surveillance system’. Their demographic characteristics, serological test results of syphilis, reporting doctors and treatment regimens were analyzed.Chi-square test was used for univariate analysis and multivariate logistic regression analysis was used to identify the influencing factors associated with CS.Results:A total of 95 CS cases were reported in NNDRS, and the reporting accuracy rate was 36.8%(35 cases). The proportion of children younger than 30 days who were reported incorrectly (73.3%(44/60)) was higher than the proportion of those were reported correctly (51.4%(18/35)), and the difference was statistically significant ( χ2=4.679, P=0.031). However, the reporting accuracy was not associated with hospital grade, doctor profession and doctor title of reporting ( χ2=0.283, 0.483 and 3.681, respectively, all P>0.05). Maternal age≤24 years (adjusted odds ratio ( aOR)=4.62), titers of non- Treponema pallidum antigen serologic test before treatment≥1∶8 ( aOR=7.56), initiation of antisyphilitic treatment during delivery ( aOR=25.43), no antisyphilitic treatment during pregnancy ( aOR=14.12), and premature delivery ( aOR=8.97) increased the risks of CS (all P<0.01). Conclusions:The reporting accuracy rate of CS is low. Young mothers, high serological titers of non- Treponema pallidum antigen before treatment, initiation of antisyphilitic treatment during delivery or no antisyphilitic treatment during pregnancy, and premature delivery are risk factors for CS. Doctors may focus on collecting this information to assist clinical diagnosis.

Objective To investigate the mechanism of effects of acitretin on the treatment of psoriasis. Methods The expression of vascular endothelial growth factor (VEGF) and micro-vascular density (MVD) were detected by immunohistochemical technique before and after treatment with acitretin in 32 patients with psoriasis. Serum level of VEGF was measured by double antibody sandwich enzyme-linked immunosorbent assay (ELISA). Results The expression of VEGF protein and MVD were significantly higher in psoriatic lesions before treatment with acitretin than those after treatment (P

AIM: To study the expression and distribution of osteopontin (OPN) in lungs and pulmonary arteries in pulmonary hypertensive rats induced by hypoxia-hypercapnia, and to explore the role of OPN in pathogenesis of pulmonary hypertension. METHODS: Forty-eight male Sprague-Dawley rats (Weight 180 g-220 g) were randomly divided into four groups: normal control group (NC), hypoxic hypercapnia 1-week,2-week and 4-week group (1HH, 2HH and 4HH). The expressions of OPN mRNA and protein in lungs and pulmonary arteries were detected by RT-PCR and immunohistochemistry. ELISA was used to detect the concentration of OPN in lung homogenates. The content of OPN in pulmonary arteries was detected by Western blotting. RESULTS: ① The mean pulmonary artery pressure (mPAP) and weight ratio of right ventricle to left ventricle and septum [RV/(LV+S)] in all hypoxic hypercapniac groups were higher than those in normal control group (P0.05). ② The expression of OPN mRNA was significantly increased in pulmonary arteries and lung tissues in hypoxic hypercapnic groups compared with normal control group (P

Abstract: The loop-mediated isothermal amplification (LAMP) technique is a technique for the specific and efficient amplification of target fragments at a constant temperature using two pairs of specially designed primers and a strand displacement activity DNA polymerase. LAMP technique is a simple, rapid, specific, sensitive and cost-effective nucleic acid amplification method, and therefore has a promising future in the field rapid detection of Mycobacterium tuberculosis and grassroots applications. In this review, the basic principles and characteristics of the LAMP technique, the main molecular markers for the diagnosis of tuberculosis, and the use of different molecular markers and various types of novel techniques in the diagnosis of pulmonary tuberculosis, extrapulmonary tuberculosis, and drug-resistant tuberculosis were described. The LAMP technique has been widely used in the diagnosis of tuberculosis with high sensitivity and specificity, but the technique still has some shortcomings. This paper reviews the progress of its application in tuberculosis in recent years and provides an outlook on its development, with a view to providing a rational research direction for rapid diagnosis of tuberculosis in a resource-limited environment.

Objective To investigate the expressions of TLR2 and TLR4 in patients with psoriasis vulgaris and the efrect of methotrexate(MTX)on them.so as to explore the therapeutic mechanism of MTX in psoriasis vulgafis.Methods Forty-three patients with psoriasis vulgaris were recruited into the study together with 30 normal human controls.Oral MTX was given to patients with an interval of 12 hours for three times per week until the control of conditions followed by 4 weeks of mainmining treatment.The dosage of MTX was 5 mg initially and decreased to 2.5 mg in the maintaining period.Flow cytometry was used to detect the expression of TLR2 and TLR4 in peripheral blood CD14~+ cells from the controls and patients at baseline,4 and 8 weeks after the beginning of treatment.Results The expression rate of TLR2 and TLR4 in CD14~+ cells was(92.6±4.3)%and(48.5±4.6)%,respectively,in untreated patients,significantly higher than that in normal controls(botll P＜0.01).A significant increase was observed in the expression rate of TLR2 and TLR4 in patients with active psoriasis compared with those with inactive psoriasis [(97.5±4.1)%vs(87.6±5.6)%,(55.3±5.8)%vs(40.7±7.1)%,both P＜0.05].Eigh weeks after the beginning of treatment with MTX.the expression rate of TLR2 and TLR4 significantly decreased to (79.6±6.7)%and(34.6±5.9)%.respectively(both P＜0.05).The psoriasis area and severity index(PASI)score had no significant correlation with the expression rate of TLR2 or TLR4(r=0.24.0.27,both P＞0.05).Conclusions TLR2,TLR4 and innate immune response mediated by both receptors play an important role in the pathogenesis of psoriasis.MTX may exert its therapeutic effect on psoriasis by inhibiting the expression of TLR2 and TLR4.

Objective To investigate the clinical phenotype, genotype and ultrastructural features in a pedigree with autosomal recessive congenital ichthyosis. Methods Patients were examined for clinical manifestation. PCR was carried out to amplify all the 15 exons and adjacent splice sites of TGM1 gene followed by bidirectional sequencing. Skin samples were taken by biopsy from the back of the proband, fixed in 3% glu-taraldehyde for transmission electron microscopy. Results The proband presented an intermediate clinical phenotype between lamellar ichthyosis (LI) and non-bullous congenital ichthyosiform erythroderma (NCIE), while his brother manifested as a collodion baby. A C551T heterozygous mutation which located in the third exon of TGM1 gene and resulted in a substitution of arginine by cysteine at codon 143 (R143C), was detected in the proband, his brother and father. Meanwhile, another heterozygous C-to-T transition at position 759 causing a substitution of serine by phenylalanine at codon 212 (S212F), was noted in the proband, his brother and mother. Electron microscopy revealed not only features of ichthyosis congenital type Ⅲ but also those of ichthyosis congenital type Ⅱ in lesions of the proband. Conclusions The patients in this pedigree carry compound heterozygous mutations, i.e. R143C, a hot missense mutation, as well as a de novo mutation S212F. The proband, who harbors mutations in the TGM1 gene, shows electron microscopic features characteristic not only of ichthyosis congenital type Ⅱ but also of ichthyosis congenital type Ⅲ.

Objective To investigate the correlation between serum interleukin-8 (IL-8 )and immunoglobulin/complement 3A (IgA/C3)and immune imbalance in Henoch-Schonlein Purpura(HSP)in children,and provide evidence and guidance for clinical therapy of HSP.Methods Serum IL-8 was measured by Enzyme-linked immunosorbent assay(ELISA),and IgA and C3 were de-tected by automatic biochemistry analyzer,and calculate the ratio of IgA/C3.Results Serum IL-8 levels were (389±5 1.68)ng/L, significantly higher than that in healthy children (P < 0.05 ).Serum IgA levels were(306.2 ± 32.21 )mg/L,significantly higher (P <0.05).Serum C3 levels were (1 14.5 ±20.7)mg/L,of no statistical difference.IgA/C3 were (3.041 ±0.508 5),significantly higher(P <0.05).There was a positive correlation between IL-8 and IgA/C3 (r =0.534,P <0.05 ).Conclusion The children of HSP has cytokine and immunoglobulin disturbance,and presents a hyperreactivity in both humoral immunity and cellular immunity. Positive correlation between IL-8 and IgA/C3 was observed.

Objective:To study the possible immune escape mechanisms of HCoV-OC43 from human dendritic cells(DC).Methods:HCoV-OC43 was isolated from clinical specimen using BSC-1 cells and identified by Real-time PCR,and the cytopathic effect was observed by phase contrast microscope.DCs were induced in vivo using hu-GM-CSF and IL-4 cytokines,and after 7 days of differentiation,DCs were infected by HCoV-OC43.The morphology of HCoV-OC43 infected DC was observed by transmission electron microscope,and the cytokines related to DC functions were detected by Real-time PCR after infection.DC proportion and function related co-stimulatory molecules were analyzed by flow cytometry.Results:In vitro HCoV-OC43 infected human DC model was successfully built.HCoV-OC43 can infect DC and generate immune response of DC in vitro,but no virus nucleonic acid could be detected in culture supernatant.The DC expression of IFN-α,IFN-β,CCL3 and CCL5 were significant decreased when infected with HCoV-OC43,but the expression of costimulatory molecules including HLA-DR,CD1c and CD86 were not affected by HCoV-OC43 infection.Conclusion:Human DC could be infected by HCoV-OC43 and generate immune response,but could not produce progeny virus.HCoV-OC43 may escape from immune response by suppressing the expression of IFN-α and other inflammatory cytokines and chemokines in DC.

One factor at a time design and first order exponential model were applied to investigate the effects of the four major factors(temperature,ionic strength,pH and osmo-regulator) in the preservation process of nitrite oxidization bacteria(NOB),and a first order decay kinetics was introduced.The result showed that the anoxic decay rate of nitrifying bacteria reduced from 0.25 to 0.015,the half life was extend to 53d at 4℃,pH7.60,ionic strength 0.035mol/kg.The protecting effect of glycerin showed obviously better than that of other cryoprotectants at 4℃.