Objective To develop a quick quantitative detecting method for point of care testing (POCT) of human serum procalcitonin (PCT) by fluorescence immunochromatographic technology.Methods Applying a double-antibody sandwich immunofluorescent assay (one antibody coated on the nitrocellulose membrane and the other antibody labeled with fluorescent micropaticles) to develop a PCT quantitative detecting kit by immunochromatography technology.The kit was used to test PCT in 472 serum samples from suspected bacterial infection patients of Guangzhou Red Cross Hospital,including 240 male and 232 female patients.The methodology and diagnostic performance were evaluated in the aspects of linearity,precision,accuracy,specificity,stability experiments and comparison with foreign PCT detecting kits.Results The report range of the PCT quantitative diagnostic kit was 0.1-125.0 μg/L The coefficient of variation (CV)values of repeat 20 tests for low,median,and high concentration control samples respectively were all less than 15％ and bias can be acceptable (P ＞ 0.05).Common interfering substances in human serum specimens such as bilirubin (2.0 g/L),triglyceride (30.0 g/L) and cholesterol (15.0 g/L) were found no significant affect on quantitative detection of PCT.The shelf time of the PCT diagnostic kit should be longer than 12 months as the relative deviation of detected concentrations of 0.5,1.0,22.0,65.0 μg/L PCTcontrol sample can be controlled less than 20％ within 14 months.Considering VIDAS BRAHMS PCT to be the standard quantitative test for PCT,472 serum samples were detected by both our kit and the control VIDAS BRAHMS PCT kit simultaneously,which showed high correlation (YVIDAS =0.180 + 1.006Xwondfo,R2 =0.988,P ＜ 0.01) and low deviation (Z =-1.6,P ＞ 0.05) without statistic significance between two methods.And the results of these two diagnostic kits showed good consistency as the area under curve of the receiver operating characteristic (ROC) of Wondfo-PCT at the three cut-off values (0.5,2.0,10.0 μg/L)were 0.997,0.994,0.998 respectively,P ＜ 0.01,using diagnostic result of the control product as standard.Kappa values were 0.899,0.905,0.973 respectively.Conclusions The method of quantitative detection of PCT by fluorescence immunochromatography for POCT was established in this study.All the observed indicators reached the clinical diagnostic requirements and can be applied for the quick detection of clinical human serum PCT.

A rapid and convenient ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) method for analysis of ribavirin and its two main kinds of metabolites, 1,2,4-triazole-3-carboxamide (TCONH2 ) and 1-β-D-ribofuranosyl-1,2,4-triazole-3-carboxylic acid (RTCOOH), in fresh eggs has been developed and validated. The sample was extracted with acetonitrile water (9 ∶ 1, V/ V), added N-hexane to move liposoluble substances, after centrifugation, the upper solvent was cleaned-up by C18 and GCB, then determined by UPLC-MS / MS coupled with Agilent ZORBAX SB-Aq column (100 mm×3. 0 mm, 1. 8 μm). Over the concentration range of 2. 0-200. 0 μg / L (0. 5-200. 0 μg / L and 5. 0-200. 0 μg / L) for ribavirin (TCONH2 and RTCOOH) in fresh egg matrix, the correlation coefficients (R2 ) of the calibration curves were above 0. 99, and the limits of detection (LODs) were 0. 54 μg / L (0. 09 and 1. 54 μg / L). The limits of quantitation (LOQs) were 1. 79, 0. 31 and 5. 13 μg / L, respectively. At three spiked concentration levels of 5. 0, 10. 0 and 50. 0 μg / L, the recoveries of ribavirin and RTCOOH ranged from 96. 1% to 99. 6%and 42. 9% to 58. 3% . And at three spiked concentration levels of 0. 5, 2. 0 and 5. 0 μg / L, the recoveries of TCONH2 ranged from 75. 9% to 106. 7% . The results of the determination in various real samples showed that the method is simple, accurate, rapid and suitable determination of ribavirin and its two main kinds of metabolites in fresh eggs.

Objective: To observe morphological changes of epithelial-mesenchymal transition in the early stage of mice renal interstitial fibrosis. Methods: Renal interstitial fibrosis was induced by unilateral ureteral obstruction(UUO) in mice. Histological and immunohistochemical methods were used to analyze pathological changes and α-SMA expression in renal tissue.Argentum hexamethylenamine staining and transmission electron microscopy were used to observe changes of the renal tubule basement membrane. Gelatin zymographic analysis was used to observe the expression of MMP2 and MMP9 in renal tissue.Results:The mice suffered from renal interstitial fibrosis were identified by histological analysis and α-SMA positive cells in renal tissue. Argentum hexamethylenamine staining and transmission electron-microscopy showed that the renal tubule basement membrane disrupted locally and renal tubule epithelial cells invaded into the renal interstitium in the early stage of renal interstitial fibrosis. Gelatin zymographic analysis showed that the expression of MMP2 and MMP9 was increased transitorily in the early stage of renal interstitial fibrosis. Conclusion: Renal tubule basement membrane disruption, renal tubule epithelial cells invasion into the renal interstitium, and the expression of MMP2 and MMP9 are involved in the development of renal interstitial fibrosis.

OBJECTIVETo study the clinicopathologic features of primitive neuroectodermal tumor (PNET) in female genital tract.

METHODSSix cases of PNET arising in female genital tract were retrospectively reviewed. The clinicopathologic features, immunohistochemical findings and EWS gene translocation study results were analyzed.

RESULTSThe age of patients ranged from 10 to 27 years (mean = 20 years). The sites of involvement included ovary (1 case), uterus (1 case), vulva (2 cases) and vagina (2 cases). The greatest diameter of the tumor ranged from 2 to 10 cm (mean = 5.4 cm). The tumor had nodular appearance and showed grayish-pink fleshy cut surface, accompanied by foci of hemorrhage and necrosis. Histologically, the tumor was composed of malignant small round cells with indistinct cell borders, hyperchromatic nuclei, dense chromatin, tiny nucleoli and scanty cytoplasm. The tumor cells were arranged in sheets or lobules. Homer-Wright rosettes were identified in 1 case. Immunohistochemical study showed that the tumor cells were positive for CD99, FLI-1 and CD56 (6/6). Focal expression of vimentin (5/6), NSE (5/6), nestin (4/6), synaptophysin (4/6), S-100 protein (2/6) and chromogranin A (1/6) was also demonstrated. EWS gene translocation was detected in 5 cases studied. Follow-up information was available in 2 patients (7 and 17 months of follow up, respectively). One of them died of tumor metastasis 17 months after diagnosis. The other patient was still alive.

CONCLUSIONSPNET arising in female genital tract is rare. It mainly involves ovary, uterus, vulva and vagina. Immunohistochemical study using a panel of antibodies and fluorescence in-situ hybridization play an important role in definitive diagnosis of this rare malignancy.

12E7 Antigen ; Adolescent ; Adult ; Antigens, CD ; metabolism ; CD56 Antigen ; metabolism ; Cell Adhesion Molecules ; metabolism ; Child ; Female ; Follow-Up Studies ; Genital Neoplasms, Female ; genetics ; metabolism ; pathology ; surgery ; Humans ; Neuroectodermal Tumors, Primitive, Peripheral ; genetics ; metabolism ; pathology ; surgery ; Ovarian Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Proto-Oncogene Protein c-fli-1 ; metabolism ; RNA-Binding Protein EWS ; genetics ; Retrospective Studies ; Translocation, Genetic ; Uterine Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Vaginal Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Vimentin ; metabolism ; Vulvar Neoplasms ; genetics ; metabolism ; pathology ; surgery ; Young Adult

OBJECTIVETo rapidly and sensitively detect the four virulence-associated factors of Streptococcus suis, a multiplex PCR was developed.

METHODSIn the process of this reaction, four distinct DNA targets were amplified. One target was based on the serotype 2 (and 1/2) specific cps gene and the others were based on Streptococcus suis mrp, epf (epf*) and sly gene, encoding the MRP, EF(EF*) and Sly proteins of Streptococcus suis. 72 isolates, which including 48 strains of Streptococcus suis and 24 strains of negative control, and 49 clinical specimens were detected by the multiplex PCR assay.

RESULTSAll PCR products were detected by electrophoresis on 1.2% agarose gels. With the 48 Streptococcus suis strains, the positive detection rates of cps2+, mrp+, epf+, epf*+ and sly+ were 16/48, 14/48, 12/48, 3/48 and 26/48,respectively. The results were confirmed by bacteriological examination. There were no specific amplification products including 49 clinical specimens and 24 negative control strains.

CONCLUSIONThe results demonstrated that multiplex PCR was a highly specific and sensitive diagnostic tool for the detection of virulence-associated factors of streptococcus suis.

Bacterial Proteins ; genetics ; Polymerase Chain Reaction ; methods ; Streptococcus suis ; genetics ; pathogenicity ; Virulence Factors ; genetics

Objective　To establish animal model for hypertrophic scar and study the characters of its morphology and collagen metabolism. Methods　A total of 64 round wounds (diameter of 6 mm each) with total skin loss were made on the ventral side of rabbit ear using a trephine. Morphology and collagen metabolism of scar wounds were studied at 14,21,35,70 and 98 days after operation, respectively. Results　There were 76% elevated scars developed (45/59 wounds) on the ventral side of rabbit ear at 21 days and 46% elevated scars disappeared (11/24) at 98 days after operation. There were numerous fibroblast proliferation and whorl-arranged collagen fibers at 21 and 35 days. The number of fibroblast decreased, but irregular-arranged fibers still presented in the elevated scars at 70 and 98 days after operation. Hydroxyproline content in elevated scars at 21 days was higher than that in normal skin (P<0.05), and at 35 days was 3 times as that in normal skin and at 98 days was also markedly higher than that in normal skin (P<0.05). Conclusion　Excessive deposition of collagen is a characteristic of hypertrophic scar in rabbits. The conversion of normal scarring to hypertrophic scarring in rabbits occurs at 14～21 days after operation. Both development and regression of hypertrophic scar in rabbit are quicker than that in human.

Objective　To establish animal model for hypertrophic scar and study the characters of its morphology and collagen metabolism. Methods　A total of 64 round wounds (diameter of 6 mm each) with total skin loss were made on the ventral side of rabbit ear using a trephine. Morphology and collagen metabolism of scar wounds were studied at 14,21,35,70 and 98 days after operation, respectively. Results　There were 76% elevated scars developed (45/59 wounds) on the ventral side of rabbit ear at 21 days and 46% elevated scars disappeared (11/24) at 98 days after operation. There were numerous fibroblast proliferation and whorl-arranged collagen fibers at 21 and 35 days. The number of fibroblast decreased, but irregular-arranged fibers still presented in the elevated scars at 70 and 98 days after operation. Hydroxyproline content in elevated scars at 21 days was higher than that in normal skin (P<0.05), and at 35 days was 3 times as that in normal skin and at 98 days was also markedly higher than that in normal skin (P<0.05). Conclusion　Excessive deposition of collagen is a characteristic of hypertrophic scar in rabbits. The conversion of normal scarring to hypertrophic scarring in rabbits occurs at 14～21 days after operation. Both development and regression of hypertrophic scar in rabbit are quicker than that in human.

Brain magnetic resonance imaging (MRI) of the elderly often reveals white matter changes (WMCs) with substantial variability across individuals.Our study was designed to explore MRI features and site-specific factors of ischemic WMCs.Clinical data of consecutive patients diagnosed with ischemic cerebral vascular disease who had undergone brain MRI were collected and analyzed.Multi-logistic regression analysis comparing patients with mild versus severe WMCs was performed to detect independent associations.Analyses of variance (ANOVAs) were used to detect regionally specific differences in lesions.We found that lesion distribution differed significantly across five cerebral areas,with lesions being predominant in the frontal lobe and parieto-occipital area.To explore WMCs risk factors,after adjusting for gender,diabetes mellitus,and hypertension,only age (P＜0.01),creatinine (P=0.01),alkaline phosphatase (ALP) (P=0.01) and low-density lipoprotein cholesterol (LDL-C) (P=0.03) were found to be independently associated with severe WMCs.Age (P＜0.001) was strongly associated with WMCs in the frontal lobe while hypertension was independently related to lesions in the basal ganglia (P=0.048) or infratentorial area (P=0.016).In conclusion,MRI of WMCs showed that ischemic WMCs occurred mostly in the frontal lobe and parieto-occipital area.The infratentorial area was least affected by WMCs.Typically,age-related WMCs were observed in the frontal lobes,while hypertension-related WMCs tended to occur in the basal ganglia and infratentorial area.

Objective: To study the biomechanical property and histocompatibility of acellular porcine fascia so as to supply a new substitute material for tissue repair. Methods: Samples of normal porcine fascia, acellular porcine fascia and normal human fascia were prepared for histological and biomechanical examination. Xenogeneic fresh porcine fascia, acellular porcine fascia and allogeneic rabbit fascia were implanted into the back of rabbit. Tissues were taken for HE staining and histocompatibility test. Results: Histological examination showed that the cellular components which elicit immune rejections had been removed in the acellular porcine fasciam, with the complete extracellular matrix reserved. Arrangement of collagen fiber was loose in the acellular porcine fascia. The biomechanical performance test of the three samples showed that there was no significant difference in the extreme tensile strength (4.47±0.54) MPa, (4.49±0.91) MPa, (4.79±1.35) MPa for acellular porcine fascia, normal porcine fascia and normal human fascia respecively, P>0.05). There was no significant difference between acellular porcine fascia extracellular matrix and porcine fascia with the cell in breaking elongation (501.83%±52.01%, 661.17%±200.28%, P>0.05). However, there was a significant difference in the breaking elongation between human fascia (98.08%±26.02%)and the other 2 kinds of porcine fascia (P<0.01). Histocompatibility test showed the local reactions of the acellular porcine fascia group were weaker than the normal porcine fascia group. Conclusions With sound preparativemethod, the acellular porcine fascia has good biomechanical property and histocompatibility and may serve as a good alternative material for tissue repair.