Objective:To develop the representative fingerprint for the quality control of placenta polypeptide injection.Methods:The chromatographic separation was performed using a Phenomenex Gemini C18 column (250 mm × 4.6 mm,5 μm) maintained at 30 ℃.0.1％ aqueous trifluoroacetic acid (Solvent A) and acetonitrile contained 0.1％ TFA (Solvent B) were used as mobile phase with a gradient elution.Detection wavelength was 280 nm with the sample injection volume of 50 μL; the flow rate was 1.0 mL/min.The fingerprints of different samples were investigated by similarity analysis.Results:Nine peaks were identified as the characteristic common peaks.The similarities of the fingerprints of the 10 batches of samples were above 0.992.Conclusion:This method showed high precision and good repeatability,and provided the basis for the improvement of the quality control of placenta polypeptide iniection.

0 05). Between the patients with high level of serum HBV-DNA and the ones with low level of serum HBV-DNA, both the quantity and positive rate of HBV-DNA in PBMC had a significant difference (P

Objective To develop real-time quantitative PCR method for measuring the amount of fetal DNA in maternal plasma and the changing patterns of fetal and maternal DNA with the pregnant progress. Methods Fifty-eight women were recruited at 7～42 weeks of gestation with singleton pregnancy determined by ultrasound scan. QIAGEN DNA kits was used to extract fetal DNA from maternal plasma. Fluorescent quantitative PCR(FQ-PCR) were applied to determine the concentration of ?-actin and SRY gene in extracted DNA. Results Thirty-seven male fetus were all identified among the 58 pregnant women (100%) and no Y signal was detected in the other 21 pregnancies with female foetus. The median level of fetal DNA in those pregnancies with male foetus was 9.08 copies/ml (3.5～12.8) in early pregnancy,45.41 copies/ml (14.38～76.5) in mid-pregnancy and 300.95 copies/ml (84～840) in late pregnancy. Conclusions The concentration of fetal DNA increases with the progress of pregnancy. Maternal plasma may be valuable in noninvasive prenatal diagnosis.

Objective To evaluate the diagnostic value of hepatitis C virus core antigen(HCV-cAg),hepatitis C virus(HCV-IgG) and hepatitis C virus(HCV-RNA) in the laboratory diagnosis of Hepatitis C.Methods HCV-cAg and HCV-IgG were detected by enzyme-linked immunosorbent assay(ELISA),HCV-RNA was detected by real-time fluorescent quantitative polymerase chain reaction(RT-PCR) in 84 suspected HCV patients and 87 healthy control subjects.Results In 84 suspected HCV patients,the HCV-IgG positive rate was 84.5%,HCV-cAg positive rate was 13.1%,HCV-RNA positive rate was 52.4%.Among 71 cases of HCV-IgG positive patients,there were 35 cases with negative HCV-RNA,the false positive rate was 49.3%.In 11 cases of HCV-cAg positive patients,there were 5 cases with negative HCV-RNA,the false positive rate was 45.5%.In 44 cases of HCV-RNA positive diagnosis of hepatitis C patients,HCV-IgG false negative rate was 18.2%,HCV-cAg false negative rate was 86.4%.The false negative rate of combined detection of HCV-cAg and HCV-IgG was 13.6%,and the true positive rate was 100.0%.Conclusion HCV-cAg and HCV-IgG have certain false negative and false positive in laboratory diagnosis of HCV,combine these two methods,or joint with HCV-RNA detection,could reduce the rate of missed diagnosis.

Objective To explore the role of ppk1 gene in E.coli CFT073 strain during urinary tract infection (UTI). Methods C57BL/6 mouse models of acute UTI with the wild-type(CFT073) and ppk1 mutant (△pk1) infected, were used to compare the bacteria colonization and inflammation induction abilities of bladder tissues. Results In the mouse models, the △pk1 strain showed a significantly lower infection rate (73.3% vs 93.3%) and lower adhesion frequency of bladder (0.01%vs 0.5%) than those of the CFT073 strain. The expression of IL-6 and TNF-αwere reduced in the bladder of △pk1 infected group (P<0.05). Hematoxylin-Eosin tissue staining showed that the damage degree of bladders in △pk1 infected mice were less serious than the CFT073 infected mice. Conclusion ppk1 gene plays an important role in E.coli colonization to bladder and the inflammation induction ability.

Objective To construct a Polyphosphate kinase 1 ( ppk1) gene deletion mutant of uro-pathogenic Escherichia coli (E.coli) CFT073, and to explore the biological properties of the mutant strain . Methods The ppk1 gene deletion strain (△pk1) was constructed based on CFT073 E.coli strain by usingλRed homologous recombination technology .A comparison analysis was conducted on adhesive and invasive abilities between CFT073 wild type strain and △pk1 strain in in vitro model of human bladder cancer epithe-lial cell 5637 .Crystal violet staining method was used to evaluate the influences of ppk1 gene deletion on biofilm formation.Results The CFT073 ppk1 deletion mutant strain was successfully constructed .Com-pared with the wild type strain ,△pk1 strain showed impaired adhesive and invasive abilities to 5637 cells. Moreover , the absorbance values of crystal violet at 570 nm at each time point of the mutant strain group were also lower than those of the wild-type strain group .Conclusion The ppk1 gene deletion mutant of uro-pathogenic E.coli CFT073 could be successfully constructed by Red homologous recombination technology and its biological properties indicates that ppk1 gene plays an important role in the pathogenesis of uropatho-genic E.coli infection through regulating the abilities of adhesion , invasion and biofilm formation .

Objective Using chemically synthesized small interfering RNA (siRNA) transfected HepG2.2.15 cells to construct a cell model in interfering hepatitis B virus (HBV) X gene, studying the inhibi-tion of HBV replication and antigen expression in vitro. Methods After transfection of HepG2.2.15 cell for 24 h, 48 h, 72 h, detecting the cell supernatant of HBsAg and HBeAg by chemiluminescence immunoassay, the cell supernatant HBxAg protein by ELISA , the HBx mRNA relative expression of transfected cell was detected by fluorescence quantitative polymerase chain reaction (PCR), the ability of cell proliferation was detected by CCK-8 assay. Results After HBx-siRNA transfected HepG2.2.15 cells, cell proliferation ability was inhibited. The cell of HBx mRNA and the cell supernatant of HBxAg expression decreased (P < 0.05); at the same time it in-hibited the expression of HBsAg and HBeAg. The suppressed peak and the inhibition rate were 66% and 58%respectively at 72 h. The fluorescence quantitative PCR confirmed that expression of HBV DNA in the super-natant was decreased. Conclusion The HepG2.2.15 cell interference model of HBV X gene has been success-fully constructed, which has an effect of inhibiting proliferation of HepG2.2.15 cells and replication and expres-sion of HBV gene in vitro.

Objective To detect the distribution of H.pylori ureA, vacA s1 gene and cagA subtype(ABC, ABD, ABAB, AAD, et al) in patients with digestive diseases in Guangzhou and investigate the relationship with the pathological findings of gastric mucosa.Methods A total of 227 randomly selected gastric mucosa from patients with digestive diseases were enrolled in the research, including 46 without pathological changes, 130 with chronic gastritis, 29 with peptic ulcer, 15 with atrophic gastritis and 7 with gastric cancer.Real-time PCR assay were used to detect Helicobacter pylori ureA gene and vacA s1 gene.EPIYA motifs in the 3′ region of cagA were amplified by conventional PCR followed by subtype sequencing. The conserved gene ureA was used to detect H.pylori infection.Results Among the 227 patients with digestive diseases, 50.7% (115/227) patients were H.pylori positive, in which 91.3%(105/115) carried vacA s1 and 78.3% (90/115) carried cagA. Four types of cagA-EPIYA subtype were detected, including ABC 17.8%(16/90), ABD 78.9%(71/90), AAD 2.2%(2/90) and ABAB 1.1%(1/90).In the non-pathological change group, 32.6% (15/46) were H.pylori positive, in which 28.3% (13/46) carried vacA s1 and 26.1% (12/46) carried cagA;in chronic gastritis group, it was 48.5% (63/130), 43.8% (57/130) and 36.2% (47/130), respectively;in ulcer group, it was 72.4% (21/29), 65.5% (19/29) and 55.2% (16/29), respectively;in atrophic gastritis group, it was 66.7% (10/15), 66.7% (10/15) and 66.7% (10/15), respectively;in gastric cancer group, it was 85.7% (6/7), 85.7% (6/7) and 71.4% (5/7), respectively.The distribution of H.pylori among the 4 groups had statistical significance (χ2=16.72;P<0.01). H.pylori was more prevalent in ulcer, atrophic gastritis and cancer group than in inflammation group and non-pathological change group (χ2=16.02;P<0.01).In patients infected by H.pylori, there was no significant difference in the distribution of vacA s1 gene as high virulence factors among non-pathological change, inflammation, ulcer, atrophic gastritis and cancer group (χ2=2.00;P=0.74), as well as cagA (χ2=3.44;P=0.49) and EPIYA subtypes (χ2=3.66;P=0.45).Conclusions H.pylori infection is significantly associated with the pathological change of gastric mucosa for patients with digestive diseases in Guangzhou, while the relationship with the pathogenicity of H.pylori with high virulence genotype is not significant.More samples and diseases reclassification are needed to make an advanced analysis of the effect of H.pylori with high virulence in gastrointestinal diseases development.

Objective To detect the expression of caveolin-1 and PY14caveolin-1 in different esophageal tissues,and study the relationships between the expressions of caveolin-1 and PY14caveolin-1 with the occurrence and development of esophageal squamous cell carcinoma.Methods The fresh surgical specimens in 60 patients with esophageal squamous cell carcinoma were collected and Western blotting were used to analyze and detect the expressions of caveolin-1 and PY14caveolin-1 in esophageal squamous cell carcinoma,adjacent esophageal tissues and normal esophageal tissues.The expression of cav-1 and PY14caveolin-1 with esophageal squamous cell carcinoma were analyzed.Results The expressions of caveolin-1 and PY14caveolin-1 in esophageal squamous cell carcinoma were much higher than that in adjacent tissues and normal esophageal tissues,and the difference between them was statistically significant(P＜0.05).Conclusion The expression of caveolin-1 and PY14caveolin-1 in esophageal squamous cell carcinoma, adjacent esophageal tissues and normal esophageal tissues decreases successively,it suggests the caveolin-1 and PY14caveolin-1 may be the key of the occurrence and development of esophageal cancer cells and they could be involved in the migration and invasion of esophageal cancer.

Morphological examination of blood cells is an important part of the hematology examination course. In order to enrich teaching resources, network of bilingual education resource was established and put into application by the Department of Hematology in Guangzhou Medical College. The repository improved teaching quality of cell morphology, and played a role in training personnel of hematology examination with solid basic skills.