Objective To explore the clinical application of treponema pallidum enzyme-linked immunosorbent assay (TP-ELISA),syphilis toluidine red unheated serum regain test (TRUST) and treponema pallidum particle agglutination assay (TPPA) for the diagnosis of syphilis,improvement of the process of sequence syphilis screening program and thus assisting clinical diagnosis and treatment of syphilis.Methods The serum samples were both screened by TP-ELISA and TRUST simultaneously.The positive was confirmed by TPPA test.Syphilis serological test results of patients were collected and retrospectively analyzed.Results The 3 618 cases out of total 82 026 serum samples were reactive by TP-ELISA,453 were reactive to TRUST,and 1 684 were confirmed positive on TPPA;thepositive rate were 4.41%,0.55%and 2.05%;The TP-ELISA,TRUST and TPPA positive conformity rate or titer were rising with the TP-ELISA S/CO and TRUST titer increasing.Conclusion There is a positive correlation of TP-ELISA S/CO among the TPPA,TRUST of positive conformity rate and titer.Using TP-ELISA and TRUST as screening,TPPA as confirmed test,the diagnostic efficiency of the process of sequence syphilis screening program can be improved.

OBJECTIVETo investigate the correlation between the ratio of serum vascular endothelial growth factor (VEGF)/endostatin (ES) and childhood acute leukemia(AL).

METHODSSerum levels of VEGF and ES were measured using ELISA in 35 children with acute AL before and after chemotherapy. The ratio of VEGF/ES was calculated. Thirty healthy children served as the control group.

RESULTSThe serum levels of VEGF (196 ± 66 pg/mL vs 29 ± 10 pg/mL) and ES (35 ± 7 ng/mL vs 19 ± 4 ng/mL) in the AL group before chemotherapy were significantly higher than those in the control group (P<0.01). The ratio of VEGF/ES in the AL group before chemotherapy was significantly higher than that in the control group (6.7 ± 3.0 vs 1.6 ± 0.7; P<0.01). In 20 children with AL who achieved complete remission, the serum levels of VEGF and ES and the VEGF/ES ratio were reduced after chemotherapy (83 ± 35 pg/mL, 27 ± 5 ng/mL, 3.1 ± 1.3, respectively; P<0.01), although the serum levels of VEGF and ES were still higher than those in the control group (P<0.01). The VEGF/ES ratio in CR patients was not significantly different from that in the control group. The serum levels of VEGF (r=0.301, P=0.045) and the VEGF/ES ratio (r=0.411, P=0.015) were positively correlated with the count of blast cells in juvenile bone marrow in 35 children with AL before chemotherapy.

CONCLUSIONSSerum VEGF and ES levels are associated with the development of childhood AL. The VEGF/ES ratio can be used to evaluate the disease progression in children with AL.

Acute Disease ; Adolescent ; Child ; Child, Preschool ; Endostatins ; blood ; Female ; Humans ; Leukemia ; blood ; drug therapy ; Male ; Vascular Endothelial Growth Factor A ; blood

Objective　The active protein of traditional Chinese medicine has anti-tumor effect, and salvia miltiorrhiza is an important anti-tumor traditional Chinese medicine. Here, the effect of Salvia miltiorrhiza lectin protein (SMLP) on the apoptosis of gastric cancer cells was studied.　Methods　SMLP expressed and purified from prokaryotic cells was used to treat the gastric cancer cells SGC-7901. The experiment was divided into the control group (untreated) and the SMLP treatment group (final concentration of 10 μmol / L of SMLP was treated for 24 h). Real-time RT-PCR and Western blot were used to detect the changes of apoptosis gene expression. Flow cytometry and Hoechst staining were applied to detect the apoptotic status. Caspase-3 and Caspase-9 activity assay kits were used to detect the apoptotic level.　Results　The result of RT-PCR showed that the mRNA level of Bax in the SMLP treatment group was significantly higher than in the control group (1.00±0.12 vs 0.67±0.10）(P<0.05). After treatment with SMLP to gastric cancer cells, the activity and expression level of cleaved Caspase-3 and Caspase-9 were increased significantly compared with the control group (P<0.05). The cell nucleus in the control group was bigger and rounder, with smooth surface and uniform staining, whilst in the SMLP-treated group, the cell nucleus became deeper with pyknosis, representing typical morphological characteristics of apoptosis. The early apoptosis level in the control group was 6.55%, and the SMLP treatment group reached 10.18%, showing an increase in the level of apoptosis.　Conclusion　SMLP expressed and purified in vitro can promote the apoptosis of gastric cancer cells, which is of great significance for further revealing the function of plant lectin and investigating the anti-tumor effect on the protein of traditional Chinese medicine.

Objective　Trihostatin A (TSA) is a histone deacetylase inhibitor of oxime salts, which has certain anti-tumor activity. This article mainly investigates the molecular mechanism of TSA inhibiting cell proliferation through p-AKT/p-mTOR pathway in gastric cancer cells.　Methods　Gastric cancer cell line-SGC-7901 were treated with TSA of different concentrations, and the inhibition rate of TSA on the cells was detected by MTT assay. The cells were divided into control group (without any treatment), TSA-treated group (200ng/ml TSA), p-AKT covering group (200 ng/mL TSA+8 μg/mL SC79) and autophagy inhibition group (5 mmol/mL 3-methyladenine+200 ng/mL TSA). The protein expression distribution of Lc3 in control and TSA group were detected by cell immunofluorescence staining. The relative expression levels of p-AKT, p-mTOR and autophagy related proteins Lc3 and P62 in control group, TSA group and p-AKT covering group were detected by Western blot. The proliferation of cells in control group, TSA group, p-AKT covering group and autophagy inhibition group were measured by EdU and cell count assay.　Results　After 24h of treatment, Lc3 in TSA group formed a large number of granular aggregates in the cytoplasm, and the fluorescence distribution changed from the initial diffuse to dense. The TSA group showed a significant reduction in green fluorescence compared with the control group in the EdU experiment. The expression levels of p-AKT in the control group, TSA group and the autophagy inhibition group were 1.78±0.19, 0.92±0.03 and 1.71±0.19, respectively, and Lc3 were 0.21±0.01, 0.79±0.06 and 0.55±0.10, respectively. Compared with the control group, the relative expression level of p-AKT in the TSA group all decreased, while the expression level of Lc3 increased (P <0.05). p-mTOR in the three groups was 0.80±0.16, 0.45±0.04 and 0.98±0.16, respectively. Compared with the control group, the relative expression level of p-mTOR in the TSA group all decreased (P <0.05). P62 in the three groups was 1.17±0.15, 0.48±0.08 and 0.77±0.10, respectively. Compared with the control group, the relative expression level of P62 in the TSA group all decreased (P<0.05). Compared with the TSA group, p-AKT, p-mTOR and P62 expression in the p-AKT covering group increased (P<0.05), while Lc3 expression decreased (P<0.05). Compared with the control group, the inhibition effect of cell growth curve was the most obvious in the TSA group, while the cell growth curve of p-AKT covering group and autophagy inhibition group showed a partial recovery compared with the TSA group.　Conclusion　TSA can promote autophagy by inhibiting p-AKt/p-mTOR pathway, thus inhibiting the proliferation of gastric cancer cells.

ObjectiveTo explore the diagnostic value of fiberoptic endoscopic examination of swallowing (FEES) combined with dye test in patients with post-stroke dysphagia and silent aspiration. MethodsFrom December, 2021 to June, 2022, 50 stroke patients in the Rehabilitation Department of Xuzhou Central Hospital were selected. They were assessed with FEES and videofluoroscopic swallowing study (VFSS), and compared. ResultsThe detection rate of aspiration was higher with FEES than with VFSS (χ² = 7.000, P < 0.05), and especially for liquid food (χ² = 4.000, P < 0.05). There was a good consistency when consuming paste food (κ = 0.941, P < 0.001) and solid food (κ = 0.779, P < 0.001). There was a good consistency in the food residue site between two methods (κ = 0.818, P < 0.001), as well as for all the three food types (κ ≥ 0.862, P < 0.001). There was no significant difference in the scores of Penetration Aspiration Scale of three food types between two methods (Z < 0.667, P > 0.05). ConclusionFEES combined with dye test can be used for evaluating silent aspiration after stroke.

PKHD1 gene mutations are found responsible for autosomal recessive polycystic kidney disease (ARPKD). However, it is inconvenient to detect the mutations by common polymerase chain reaction (PCR) because the open reading frame of PKHD1 is very long. Recently, long-range (LR) PCR is demonstrated to be a more sensitive mutation screening method for PKHD1 by directly sequencing. In this study, the entire PKHD1 coding region was amplified by 29 reactions to avoid the specific PCR amplification of individual exons, which generated the size of 1 to 7 kb products by LR PCR. This method was compared to the screening method with standard direct sequencing of each individual exon of the gene by a reference laboratory in 15 patients with ARPKD. The results showed that a total of 37 genetic changes were detected with LR PCR sequencing, which included 33 variations identified by the reference laboratory with standard direct sequencing. LR PCR sequencing had 100% sensitivity, 96% specificity, and 97.0% accuracy, which were higher than those with standard direct sequencing method. In conclusion, LR PCR sequencing is a reliable method with high sensitivity, specificity and accuracy for detecting genetic variations. It also has more intronic coverage and lower cost, and is an applicable clinical method for complex genetic analyses.
DNA Mutational Analysis ; Exons ; genetics ; Genetic Testing ; Genotype ; Humans ; Introns ; genetics ; Mutation ; Polycystic Kidney, Autosomal Recessive ; diagnosis ; genetics ; Polymerase Chain Reaction ; Receptors, Cell Surface ; genetics ; isolation & purification ; Sequence Analysis, DNA

OBJECTIVE: To explore the role of N~6-methyladenosine(m~6A) catalytic enzymes(methyltransferases and demethylases) in cadmium-induced oxidative damage in human renal epithelial cells(HK-2 cells), and to analyze the correlation between nuclear factor-erythroid 2-related factor 2(NRF2) and m~6A catalytic enzymes. METHODS: i) HK-2 cells in logarithmic growth phase were randomly divided into control group and 6 cadmium sulfate treatment groups, then treated with 0, 2, 4, 8, 16, 32 and 64 μmol/L cadmium sulfate solution for 24 hours. The cell survival rates were detected by CCK-8 assay, and the appropriate doses of cadmium sulfate were selected for subsequent experiments. ii) HK-2 cells in logarithmic growth phase were randomly divided into control group and low-, medium-, and high-dose groups, and treated with 0, 4, 8, and 16 μmol/L cadmium sulfate solution respectively for 24 hours. Subsequently, the levels of reactive oxygen species(ROS) were detected by fluorescence probe. The mRNA expression of NRF2, the m~6A methyltransferases such as methyltransferase like proteins(METTL) 3, METTL14, METTL16 and the m~6A demethylases such as fat mass and obesity associated protein(FTO), AlkB family of nonheme Fe(Ⅱ)/α-ketoglutarate(α-KG)-dependent dioxygenases 5(ALKBH5) were determined by real-time polymerase chain reaction. RESULTS: i) The survival rate of HK-2 cells was more than 60.00% and lower than that of the control group(P<0.05) after the cells were stimulated with 16 μmol/L of cadmium sulfate. Therefore, 4, 8 and 16 μmol/L of cadmium sulfate were selected as the stimulation concentrations in the follow-up experiments. ii) The relative expression of NRF2, METTL3, METTL14 and METTL16 in HK-2 cells in low-dose group increased(all P<0.05), while the levels of ROS and the relative mRNA expression of NRF2, METTL3, METTL14, METTL16 and FTO in HK-2 cells in medium and high-dose groups increased(all P<0.05) when compared with the control group. There was no significant difference in the expression of ALKBH5 mRNA among these 4 groups(P>0.05). In the correlation analysis, NRF2 mRNA expression was positively correlated with the mRNA expression of METTL3 and METTL16 [Pearson correlation coefficient(r) = 0.61 and 0.66, respectively, all P<0.05]. There was no correlation between NRF2 mRNA expression and METTL14, FTO and ALKBH5(r=0.53, 0.48, and 0.01 respectively, all P>0.05). CONCLUSION: Cadmium sulfate may increase intracellular ROS level, up-regulate NRF2 expression and activate NRF2 signaling pathway as well as enhance the expression of METTL3 and METTL16 in HK-2 cells, thus increasing intracellular oxidative damage and decreasing the cell survival rate.