Objective To evaluate the influence of Eperythrozoon infection on human and mouse erythrocytes and to explore the pathogenesis of Eperythrozoonosis. Methods The specific gene fragment of Eperythrozoon was detected by polymerase chain reaction (PCR) from the venous blood samples of five patients infected with Eperythrozoon. The complement receptor type I (CD35) expression on erythrocytes of these five patients was determined by flow cytometry. Thereafter, the Eperythrozoons were purified from human samples and injected into mice through the tail veins. Blood smear microscopy, PCR and transmission electron microscopy were used to assure the successful infection. The hematological indicators of human and mice, such as red blood cell (RBC) count,hemoglobin (Hb) content, hematocrit and superoxide dismutase (SOD) were evaluated. All results were analyzed by t test. Results More than 80% of treated mice were confirmed to be infected with Eperythrozoon successfully. A fragment of 801 bp specific gene of Eperythrozoon was detected by PCR in samples from both infected patients and infected mice, which were not detected in samples from healthy control people or control mice. CD35 was highly expressed on the erythrocytes of infected patients, but not expressed on the erythrocytes of infected mice. Both RBC counts and Hb content dramatically decreased in infected patients and infected mice. Hematocrit and the activity of SOD also slightly decreased in infected patients and infected mice. Conclusions Eperythrozoon can spread between human and mice and destroy erythrocyte structure. Eperythrozoon can upregulate CD35 expression in human, but there is no CD35 expression in mice.

Humans ; Infant, Newborn ; Microbiological Techniques ; Sepsis ; blood ; microbiology

Adenocarcinoma, Clear Cell ; metabolism ; pathology ; surgery ; Adult ; Carcinoma ; metabolism ; pathology ; Carcinoma, Mucoepidermoid ; metabolism ; pathology ; Carcinoma, Renal Cell ; metabolism ; pathology ; secondary ; Diagnosis, Differential ; Humans ; Keratins ; metabolism ; Male ; Nasal Cavity ; Nose Neoplasms ; metabolism ; pathology ; surgery ; S100 Proteins ; metabolism

BackgroundChoriodal neovascularization is an important ocular manifestation of angiogenesis in eyes,which derives from the choroid capillaries.Recent studies have found that complement activation is playing a key role in the laser-induced CNV.Because of the key position of CFB in the alternative pathway,bytargeting CFB and blocking the alternative pathway may provide an approach to observe the role of this alternative pathway in the generation of CNV.Objective This study was to investigate the inhibitory effect of reconstructed complement factor B (CFB)-small interfering ribonucleicacid(siRNA)on choroidal neovascularization (CNV)and its mechanism. Methods Experimental CNV was induced by laser photocoagulation in 96 eyes of 48 clean Brown Norway rats.The rats were randomly divided into 4 groups.25,50 and 75 μg B factor siRNA were injected via caudal vein on 1 day,3,5 days after photocoagulation in different dose groups,and normal saline solution was injected at the same way in experimental control group.Other 12 normal rats were used as blank control group.Fundus fluorescein angiography(FFA) was performed on 3,7,14,21,28 days after injection of CFB-siRNA and CNV was scored.The expressions of vascular endothelial growth factor(VEGF) and factor Ⅷ in choroid were detected by immunochemistry.The expressions of CFB-siRNA,VEGF,transforming growth factor β2( TGF-β2 )proteins in choroid were determined using immunochemistry in 7,14,21,28 days,and the expressions of mRNA of CFB-siRNA,VEGF,TGF-β2 were examined by reverse transcription polymerase chain reaction(RT-PCR). ResultsFFA revealed that the CNV rates in various doses of CFB-siRNA groups were significant lower than those of experimental control group in various time points(P＜0.05),and those in 75 μg B factor siRNA were decreased in comparison with 25 μg B factor siRNA (P＜0.05).Immunochemistry showed that the intensities of the VEGF and factor Ⅶ expression in various doses of CFB-siRNA groups were weaker than the blank control group ( P ＜ 0.05 ).Compared with the control group,the expression of CFB reduced in 7 days,and then approached to the level near the control group.Fourteen to twenty-one days after injection of CFB-siRNA,VEGF and TGF-β2 depressions in different doses of CFB-siRNA groups were lower than blank control group( P＜0.05 ).CFB expression in choroid showed the lower levels in CFB-siRNA injection group compared with blank control group in from 7 through 21 days (P＜0.05).RT-PCR displayed the gradual increase of CFB mRNA and curve-like changes of VEGF and TGF-β2 with time prolong. Conclusions Recombinated CFB-siRNA can effectively inhibit laser-induced CNV by down-regulating the expression of VEGF and factor Ⅷ.Alternative pathway of complement plays an important role in the production of CNV.

Albuminuria ; drug therapy ; Diabetes Mellitus, Type 2 ; complications ; drug therapy ; Diabetic Nephropathies ; drug therapy ; Humans ; Tetrazoles ; therapeutic use

Preparative HPLC was used to prepare ferulic acid, senkyunolide I and senkyunolide H from Ligusticum chuanxiong. The separation was conducted on a Shim-Pack Prep-ODS (20.0 mm x 250 mm, 5 microm) column with the mobile phase of methanol-0.2% glacial acetic acid (50:50)at the flow rate of 5 mL x min(-1). The detection wavelength was 278 nm, and the purity of each compound was detected by HPLC analysis. Spectral data analyses including UV, ESI-MS and NMR were used to identify their structures. This method is simple, fast, which is suitable for preparation of standard reference of ferulic acid, senkyunolide I and senkyunolide H from L. chuanxiong and can meet the requirement of new drug research and development.
Benzofurans ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Coumaric Acids ; chemistry ; isolation & purification ; Ligusticum ; chemistry

OBJECTIVEIn this study, we discussed the consistency and correlation of HBV serological indexes between neonates' venous blood and cord blood whose mothers had chronical HBV infection, as well as the correlation of thoses indexes with the mothers'.

METHODChronically HBV infected mothers who were postive of both HBsAg and HBeAg and also had a HBV DNA virus load above 10(5) copies/ ml and their infants were enrolled. The mothers' venous blood were collected before delivery. The neonates' cord blood were collected at birth after removal of contaminants and disinfected with alcohol on the cord's surface, and the venous blood were collected before hepatitis B virus immune globin(HBIG) and hepatitis B vaccine were given. The levels of HBsAg, anti-HBs, HBeAg and anti-HBeAg were tested with Abbott microparticle chemiluminescence method (Abbott Laboratories, Abbott Architac i2000). HBV DNA quantification were tested by COBAS TagMan real-time PCR Assay.

RESULTS383 mothers and their infants were enrolled. The positive rates of HBsAg in cord blood and venous blood were 61.2% and 63.9%. The positive rates of HBeAg level in cord blood and venous blood were 83.2% and 83.5%. The positive rates of HBV DNA level in cord blood and venous blood were 56.0% and 59.4%. The state of HBsAg, HBeAg and HBV DNA in cord blood and venous blood were consistency, and significant correlation was observed in their levels with correlation coefficients of 0.766, 0.857, and 0.692, respectively (P < 0.000). Significant correlation of the HBeAg levels were observed between mothers' venous blood and neonates' venous blood, as well as neonates' cord blood with correlation coefficients of 0.362 and 0.352 (P < 0.000). However, there was no significant correlation of HBsAg levels between them (r = 0.023, P = 0.785; r = 0.04, P = 0.604).

CONCLUSIONSThe HBV serological index of neonate's cord blood could reflect the HBV serological indexes in venous blood because of the good correlation and consistency between them.

DNA, Viral ; blood ; Female ; Fetal Blood ; virology ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; virology ; Humans ; Infant, Newborn ; Pregnancy ; Pregnancy Complications, Infectious ; virology ; Veins

Aspergillosis ; diagnosis ; microbiology ; Aspergillus fumigatus ; genetics ; isolation & purification ; DNA, Fungal ; genetics ; isolation & purification ; Dermatomycoses ; diagnosis ; microbiology ; Eye Diseases ; diagnosis ; microbiology ; Female ; Humans ; Middle Aged ; Polymerase Chain Reaction

This article reports the clinical features and endocrine and metabolic features of 4 children with Prader-Willi syndrome (PWS). All the patients were female and aged 6-12 years at diagnosis. All of them had clinical manifestations of obesity, unusual facies, developmental retardation, and intellectual disability. Genetic detection showed that 2 patients had paternal deletion of the 15q11.2-q13 region, one patient had maternal autodiploid in the 15q11.2-q13 region, and one patient had no abnormality in the 15q11.2-q13 region. All patients had varying degrees of endocrine and metabolic disorders: 2 patients had short stature, among whom one had delayed appearance of secondary sex characteristics and the other one had type 2 diabetes; one patient had insulin resistance and no mammary gland development; one patient had a body height of P-Pand precocious puberty. Patients with PWS have various endocrine disorders, so long-term endocrine follow-up and management is very important.
Child ; Child, Preschool ; Endocrine Glands ; physiopathology ; Female ; Glucose Tolerance Test ; Humans ; Prader-Willi Syndrome ; genetics ; physiopathology

The formalin test is a commonly used animal model of acute and tonic pain. However, the molecular targets of formaldehyde (FA, the main ingredient of the formalin solution) on primary nociceptor cells remain controversial. In this report, the effects of FA on electrophysiologically-identified primary nociceptor cells were evaluated in vitro and the roles of the vanilloid receptor TRPV1 in FA-produced activation of primary nociceptors were also examined at both cellular and behavioral levels. Of 92 acutely dissociated dorsal root ganglion (DRG) cells recorded by current patch-clamp technique, 34% were discharged by FA application with the mean onset latencies of the first action potential (AP) being (367.34+/-32.96) s. All the FA-sensitive cells were identified as nociceptor cells by their distinguishable features of AP including longer duration, existence of a hump (a shoulder or inflection) on the repolarizing phase, and longer after-hyperpolarization of APs. Co-application of capsazepine (CPZ), a competitive antagonist of TRPV1 receptors, could block FA-evoked firing with partial inhibition on the membrane depolarization of all cells tested. Of another 160 cells examined by confocal calcium imaging, 32% were shown to respond to FA with an intracellular Ca(2+) rise. Of 51 FA-sensitive cells, 67% were suppressed by CPZ, suggesting partial involvement of TRPV1 in mediation of the FA-evoked intracellular Ca(2+) rise. Under voltage-clamp mode, 41% of DRG cells were evoked to give rise to inward current with the remaining 59% being unchanged. In separate experiments on the other 56 FA-sensitive cells, concentration-dependent increase in the FA-evoked current amplitude was demonstrated. In comparison with controls, the FA-evoked inward current could be significantly suppressed by CPZ that was further enhanced by HC-030031, a TRPA1 selective antagonist. Finally, local effects of CPZ were confirmed in the formalin test and it was shown that the formalin-induced paw flinches were strongly suppressed by CPZ in phase 1 but with phase 2 being significantly suppressed only during 25-55 min. It is therefore concluded that FA can directly activate a subpopulation of primary nociceptor cells and the FA-induced AP discharges are likely to contribute mainly to phase 1, but not phase 2 of the formalin-induced nociception. The activation of primary nociceptor cells by FA is likely to be mediated, at least in part, through TRPV1 and/or TRPA1 receptors.
Acetanilides ; pharmacology ; Action Potentials ; Animals ; Capsaicin ; analogs & derivatives ; pharmacology ; Formaldehyde ; pharmacology ; Ganglia, Spinal ; physiology ; Nociceptors ; physiology ; Pain ; physiopathology ; Pain Measurement ; Patch-Clamp Techniques ; Purines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; TRPV Cation Channels ; physiology