الملخص
Objective:This study evaluates the performance of chemiluminescence assay, which is designed to detect Hepatitis D Virus (HDV) Immunoglobulin G (IgG) antibodies.Methods:A comparative analysis was conducted among chemiluminescence anti-HDV IgG reagent, the magnetic particle-based domestic reagent A and domestic reagent B, and the Robo Gene HDV RNA kit, using 1909 HBsAg-positive plasma samples. This comparison aimed to delineate clinical specificity and detection accuracy. The anti-HDV IgG reagent precision was assessed at three different concentration levels following the Clinical Laboratory Standards Institute EP5-A2 guidelines. The specificity of the assay was validated using 200 HAV IgM positive, 545 HBsAg-positive but anti-HDV IgG-negative, 350 anti HCV positive plasma samples and 200 healthy human blood samples. Additionally, a concordance study was conducted with 545 HBsAg-positive and 37 anti-HDV IgG-positive plasma samples, comparing the anti-HDV IgG reagent against reagent A.Results:1 909 HBsAg-positive plasma samples were tested using 3 anti HDV IgG reagent and 1 HDV RNA reagent, 19 samples were identified as anti-HDV IgG-positive. The anti-HDV IgG demonstrated superior accuracy and specificity. The assay exhibited excellent precision, with intra-assay coefficient of variation (CV) values ranging from 1.57% to 4.30%, and inter-assay CV values between 1.71% and 4.67% for detecting samples at high, medium, and low concentration levels. Concordance with Reagent A showed consistent results in both positive and negative detections.Conclusion:In this study, the anti-HDV IgG reagent (chemiluminescence method) displayed outstanding specificity in detecting clinical samples and exhibited a high conformity rate with commercialized reagents, making it potentially suitable for screening anti-HDV IgG in HBsAg-positive samples.
الملخص
Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.
الملخص
OBJECTIVE To analyze the chemical constituents and components absorbed into plasma of the extract of Ardisia crenata and to elucidate its possible pharmacodynamic material basis. METHODS Overall， 12 rats were randomly assigned to the blank group （n＝6） and A. crenata group （n＝6） by the paired comparison method. The drug was administered once daily in the morning and afternoon for three days. Serum samples were prepared from serum after redosing on 4th day. The UPLC-QE-HF-MS/ MS was used to analyze and identify the chemical constituents in A. crenata extract and serum samples. Compound Discoverer 3.0 was employed for retention time correction， peak identification， and peak extraction. According to the secondary mass spectrometry information， the Thermo mzCloud online and Thermo mzVault local databases， referring to the relevant literature and control quality spectrum information were used to preliminarily identify the chemical constituents and components absorbed into plasma of A. crenata. RESULTS A total of 34 compounds were identified from the extract of A. crenata， mainly coumarins， flavonoids， organic acids， amino acids， including bergenin， quercetin， gallic acid， L-pyroglutamic acid， etc. Besides， 5 components absorbed into plasma were identified from serum samples: L-pyroglutamic acid， syringic acid， bergenin， cinnabar root saponin A， and mycophenolic acid. CONCLUSIONS L-pyroglutamic acid， syringic acid， bergenin， cinnabar root saponin A， and mycophenolic acid may act as the pharmacodynamic material basis of A. crenata.
الملخص
OBJECTIVE：To identif y and analyze the flavonoids and coumarins in Radix Ardisiae from different sources. METHODS：UPLC-QE-HF-MS/MS was adopted. The determination was performed on Zorbax Eclipse-C 18 column with mobile phase consisted of acetonitrile- 0.1％ formic acid solution （gradient elution ）at the flow rate of 0.3 mL/min. The column temperature was 30 ℃，and the temperature of injector was 4 ℃. The sample size was 2 µL；ESI source was applied in negative and positive scanning ion mode ，the heater temperature was 325 ℃，the sheath gas pressure was 45 arb，the auxiliary gas pressure was 15 arb，the purge gas pressure was 1 arb，the electrospray voltage was 3.5 kV，the capillary temperature was 330 ℃， S-lens RF level was 55％，scan mode was first-order full sca m/z 100-1 500，data-dependent secondary mass spectrometry scanning （dd-MS2，Top N ＝10），the resolution was 70 000 （first mass spectrometry ） ， 17 500 （secondary mass spectrometry），the collision mode was high-energy collision dissociation. Through retrieving foreign and domestic databases as ChemSpider ，mzCloud，mzVault，PubChem，the structure of the compound was identified on the basis of related literatures and reference data ，and the conten ts were compared. RESULTS & CONCLUSIONS：A total of 47 components were separated from Radix Ardisiae of 3 kinds of sources as Ardisia crenata Sims，A. crispa（Thunb.）A. DC. ，A. crenata Sims var . bicolor （Walk）C. Y. Wu et C. Chen. A total of 17 flavonoids were identified ，including 9 flavonols （quercetin 3-O-rhamnoside-7-O-glucoside， myricetin， rutin， mauritanin， kaempferol， quercetin， isorhamnetin， quercetin，mearnsitrin），3 flavan-3-ols [（-）-epigallocatechin，catechin，epigallocatechin gallate ）2 dihydroflavonoids [fustin ， eriodictyol] and 3 other types [ 3-（2，3-dihydro-benzo[1，4]dioxin-6-yl）-7-hydroxy-2-trifluoromethyl-chromen-4-one，methadone， oriciacridone F] ，10 coumarins {bergenin ，（[ 7-hydroxy-4-methyl-2-oxo-2H-chromen-6-yl）oxy]acetic acid ，[7-（carboxymethoxy）- 4-methyl-2-oxo-2hydroxychromo-3-yl]acetic acid ，4，9-dihydroxy-7H-furo[3，2-g]chromen-7-one，6，7-dihydroxy-4-methylcoumarin， esculetin，fraxetin，7，8-dihydroxy-4-methylcoumarin，4-methylumbelliferyl glucuronide ，scoparone}. Results of content analysis showed that in flavonoids and coumarins ，there were 5 common components in Radix Ardisiae from 3 kinds of sources ，i.e. bergenin（peak 2），[7-（carboxymethoxy）-4-methyl-2-oxo-2-hydroxychromo-3-yl] acetic acid （peak 5），methadone（peak 16）， quercetin（peak 18），oriciacridone F （peak 26）；the contents of common components were significantly different. In addition to 5 common components ，there were 22 different chemical components ，which were compounds corresponding to peaks 1，3，4， 6-15，17，19-25 and 27，respectively. Among them ，compounds corresponding to peaks 3，6，8 and 23 were only found in A. crenata Sims var. bicolor（Walk）C. Y. Wu et C. Chen ；compounds corresponding to peaks 12-15，19 were only found in A. crispa （Thunb.）A. DC. UPLC-QE-HF-MS/MS method can efficiently ，accurately and quickly identify the flavonoids and coumarins in Radix Ardisiae from different sources.
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Objective To investigate the effect of bevacizumab combined with chemotherapy on the expression of EGFR and HER in the patients with metastatic colorectal cancer .Methods Sixty patients with metastatic colorectal cancer treated in this hos-pital from January 2013 to October 2016 were selected and randomly divided into the experiment group (bevacizumab combined with FOLFOX-6 chemotherapy regimen) and control group (FOLFOX-6 chemotherapy regimen) 30 cases in each group .The curative effects were observed in the two groups .The levels of serum HER and EGFR were detected by enzyme-linked immunosorbent assay (ELISA) .Results The total effective rate after treatment in the experiment group was 53 .33% ,which in the control group was 33 .33% ,the difference was statistically significant (P< 0 .05) .After treatment ,the levels of serum HER and EGHR in the two groups were decreased to some extent ,moreover the experiment group had more decrease than the control group ,the difference be-tween the two groups was statistically significant (P<0 .05) .The occurrence rate of adverse reactions had no statistical difference between the two groups (P>0 .05) .Conclusion Bevacizumab combined with chemotherapy has good effect in the patients with metastatic colorectal cancer .
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Objective To investigate the changes of erythrocyte parameters and the value of differential diagnosis in pregnant women with β-mediterranean anemia.Methods A total of 300 pregnancy women from July 2014 to December 2015 in Center Hospital of Longgang were recruited in this study,100 pregnant women with β-mediterranean anemia in β-mediterranean anemia pregnancy group,100 healthy pregnant women in normal pregnancy group,100 pregnant women with iron deficiency anemia in iron deficiency anemia pregnancy group.Mean red cell volume(MCV),mean erythrocyte hemoglobin(MCH),reticulocyte percentage(Ret%) were detected and compared in the three groups.Results Compared with the normal pregnancy group and iron deficiency anemia pregnancy group,the MCV,MCH significantly reduced,and Ret% significantly rised in the β-mediterranean anemia pregnant group,the differences were significant(P<0.05).The best cut-off value of Ret% was 1.7% in differential diagnosis of β-mediterranean anemia pregnancy and iron deficiency anemia pregnancy,the sensitivity was 63.00%,the specificity was 74.00%,the area under of receiver operating characteristic curve was 0.841.The sensitivity of joint detection including MCV,MCH and Ret% in differential diagnosis of β-mediterranean anemia pregnancy and iron deficiency anemia pregnancy was 84.00%,the specificity was 90.00%.Conclusion MCV,MCH and Ret% in pregnancy women with β-mediterranean anemia changes significant compared with normal pregnancy group and iron deficiency anemia pregnancy group,the joint detection including MCV,MCH and Ret% could significantly improve the differential diagnosis of β-mediterranean anemia and iron deficiency anemia in pregnancy women.
الملخص
Objective To investigate the expressions of cytokeratin/keratin 4 in the buccal tissue of oral submucous fibrosis at the early,middle and advanced stages,explore which role keratin4 (K4) plays in the process and development of oral submucous fibrosis (OSF),and provide evidence for K4 being a promising biomarker to evaluate the development and prognosis of OSF.Methods Ten cases of normal tissues,and 10 cases of OSF tissues with typical early,middle and advanced stages,were selected,respectively.Detect the expression of K4 in the tissue mentioned above through immunohistochemistry and Westem blot.The data was analyzed by statistical means.Results The results of immunohistochemistry showed that K4 was mainly located in cytoplasm,and positive cells with brownish yellow granules were seen in whole epithelial layer of the normal mucosa.The expression of K4 was lower at all stages of OSF than that at the normal tissue with statistical significance (P ＜0.05).With the aggravation of OSF,the expression of K4 was decreased,difference between early and advanced stage was found to be statistically significant.The results of Western blot also showed that the expression of K4 was lower than that of early,middle and advanced stages of OSF (P ＜ 0.05).With the aggravation of OSF,the expression of K4 was decreased,but the differences between them had no significance (P ＞ 0.05).Conclusions The expression of K4 in OSF tissue of early,middle and advanced stages were decreased compared to normal tissue,respectively.It suggests that K4 might play and important role in the initiation and development of OSF.