Objective To establish a new approach for quantitative detection of VAD1 mRNA in cryptococcus neoformans by RT-FQ-PCR, and evaluate the treatment efficacy of CNM. MethodsThe primers and TaqMan probe were designed according to the published sequence of VAD1 mRNA (GenBank),and RT-FQ-PCR method to detect VAD1 mRNA was established. Cerebrospinal fluid from 25 CNM patients and 30 controls were detected and sensitivity and specificity of the method were evaluated. VAD1 mRNA concentration in cerebrospinal fluid from both acute phase and recovery phase of 25 CNM patients were also detected and significance of CNM treatment efficacy with VAD1 mRNA analysis was evaluated. Results Correlation coefficient of standard curve was - 0. 997 9 in detection of VAD1 mRNA by RT-FQ-PCR, and the detection limit was 101 copies/μl. The intra CV of plasmid standard for high, medium and low concentrations were 0. 65% ,0. 89% and 1.23% respectively, the sensitivity of cryptococcus neoformans detection by RT-FQPCR was 96% (24/25) ,while specificity was 100% (30/30). VAD1 mRNA concentration in acute phase were significant higher than that in recovery phase (3. 042 ±0. 906 vs 2. 187 ±0. 665 ,t =4. 583 ,P ＜0. 01).Conclusions The established RT-FQ-PCR method for the detection of VAD1 mRNA is provided with sound sensitivity, specificity and reproducibility, which might be fit for the detection of VAD1 mRNA. The expression level of VAD1 mRNA is relevant with the treatment efficacy of CNM.

50 mm/h were detected in all the patients. Decreasing C 3 in 6 patients and C 4 in 5 were found. Conclusions SLE is one of the general conditions causing retinal vein occlusion. Visual acuity and barrier of retinal vein and capillary are damaged seriously in patients with retinal vein occlusion caused by SLE, which may be accompanied with other ocular or general lesions. It is suggested that retinal vein occlusion is relative with SLE activity.

Objective To investigate the clinical characteristic of ocular fundus complications in systemic lupus erythematosus (SLE). Methods In 25 cases of SLE with the ocular fundus complications, the ocular fundus, the other ocular tissues, general lesions,and antinuclear antibody (ANA), anti-double-stranded DNA(anti-dsDNA), complement 3 (C 3), complement 4 (C 4)and erythrocyte sedimentation rate(ESR) were analyzed retrospectively. Results In the 25 cases, "classic" SLE retinopathy in 15 (25 eyes), retinal vein occlusion (RVO) in 9 (12 eyes), RVO combined with retinal artery occlusion in 1 (2 eyes), exudative retinal detachment in 1 (2 eyes), vitreous hemorrhage combined with neovascular glaucoma in 1 (1 eye), and optic disc edema except RVO in 3 (6 eyes) were found. Nine cases accompanied with other ocular signs and 21 with general lesions. Positive ANA and anti-dsDNA and elevated ESR in all of the patients, decreasing C 3 in 19, and C 4 in 17 were found. Conclusions SLE can cause serious ocular fundus complications accompanied with other ocular signs. Regular ophthalmic examination should be performed on the patients with SLE to detect and treat the ocular complications promptly.

Objective To observe the effects of aminoguanidine and Vitamin C on lipid of streptozotocin-induced didbetic rats.Methods Diabetic nephropathy rats were induced by intraperitoneal injection of streptozotocin.Fifty Sprague-Dawley(SD) rats were randomly divided into normal control group,model group,Vitamin C group,aminoguanidine group and Vitamin C associated with aminoguanidine group.All the rats were treated for 16 weeks.During and after the treatment,the general state of the rats was observed and the levels of blood sugar,triglyceride,total cholesterol,low density lipoprotein,high density lipoprotein,glycosylated hemoglobin,and glycated low density lipoprotein in different groups were detected.Results The rats in the four groups suffered diabetes mellitus and renal function lesion.Aminoguanidine and Vitamin C could improve the general state of the rats,but had no effect on the blood sugar.Vitamin C could decrease the levels of triglyceride,serum cholesterol,low density lipoprotein,glycosylated hemoglobin,and glycated low density lipoprotein,and increase the level of high density lipoprotein.There was a synergistic effect with the combination of the two drugs. Conclusion Aminoguanidine and Vitamin C have no effect in the decrease of blood sugar level but have a certain effect on regulating blood lipid.There is a synergistic effect with the combination of the two drugs.

Objective To analyze the characteristics of blood loss for laboratory test of critically ill premature infants,and to seek feasible measures to reduce the blood loss.Methods Two hundred and forty-six cases of critically ill premature infants admitted to the neonatal intensive care unit from April 2012 to April 2013 were analyzed the blood loss for test during the hospitalization and the clinical features of blood loss with different gestational age,different weight within the first four weeks after admission.Then the application of blood loss according to test category was described.The blood volume demanded in theory was determined by the formula B =5 (∑ s + 0.1),then calculated the phlebotomy overdraw on the basis of the practical blood loss and analyzed the characteristics of overdraw per patients per day in first two weeks after admission.Results Among 246 patients,The median blood loss figure was 25.57 ml for each infants with the range between 7.10 ml ～ 119.20 ml,and the blood loss concentrated in first four weeks,which showed a decreasing trend with time.There was a statistically significant difference(P ＜0.05)that the smaller gestational age,the lower birth weight,the more daily blood loss per patient per day in first two weeks,but no significant differences(P ＞ 0.05) between the third and fourth week.The largest proportion of the blood samples was used for clinical chemical tests(31.49％),followed by blood gas analysis (19.03％),immunoassays (12.69％),blood cultures (12.63％),hematology (12.28％).The practical blood loss was about twice times of blood volume demanded for tests in theory,which the median was 7.8 times to the latter(25.57 ml vs 3.26 ml).It showed statistically significant difference(P ＜0.05) between blood overdraw per patients per day in the first week and the second week.Compared with different gestational age and birth weight,the difference of overdraw was also statistically significant(P ＜0.05).According to test category,blood culture was the most significant samples of phlebotomy overdraw,followed by biochemical,other,blood gas analysis,the percentage was 76％,64％,45％ and 41％ respectively.Conclusion The blood loss for laboratory test and the phenomenon of blood waste is serious in critically ill preterm infants.The smaller the gestational age is,the lower the weight is,the amount of blood loss and phlebotomy overdraw are more significantly.Biochemical and blood gas analysis are the main items of blood loss.

Background Attack of primary acute angle-closure glaucoma (PAACG) often cause damage of retinal function.To measure the structural change after acute angle-closure glaucoma attack is of an important clinical significance.Flourier domain OCT(FD-OCT) is available.Objective This study was to measure and compare the thickness of peripapillary and macular retinal nerve fiber layer (RNFL) and macular volume in PAACG patients and normal subjects.Methods A case-controlled study was designed.Twenty-five eyes after acute angle-closure glaucoma attack and 25 fellow eyes were collected from April,2011 to February,2012 in Affiliated Eye Hospital of Wenzhou Medical College,and 25 eyes of normal subjects with the matched age and gender were enrolled at the same period.The thicknesses of RNFL and macula as well as the volumes of macula were measured by FD-OCT in all the eyes 2 weeks after acute angle-closure glaucoma attack.The outcomes were compared among the attacked eyes,fellow eyes and normal control eyes.Results The average thicknesses of RNFL were (125.72 ± 28.57),(108.36 ±9.31) and (106.10± 10.97)μm in the attacked eyes,fellow eyes and normal control eyes,showing a significant increase in the attacked eyes (all at P ＜ 0.05).The RNFL thickness values in the superior,interior and nasal quadrants at optical disc were significantly higher in the attacked eyes than those of the fellow eyes and normal control eyes (all at P＜0.05),but no significant differences were found in temporal and temporal-lower quadrants among the three groups (P=0.081,0.766).The thickness values of macular area were (283.72 ± 18.33),(280.28 ± 16.85) and (289.14±10.60)μm in the attacked eyes,fellow eyes and normal control eyes,with no significant difference among them (Fgroup =2.048,P =0.136),and fellow eyes were significantly reduced in comparison with the normal control group (P＜0.05),and those in the attacked eyes was not significantly reduced in comparison with the normal control group (P =0.224).The macular volumes were (5.589±0.355),(5.532±0.325) and (5.720±0.241)mm3in the attacked eyes,fellow eyes and normal eyes,without statistically significant difference among them (Fgup =1.027,P=0.363).The macular thickness values were lower in the outer race than those of the inner race and followed the pattern of nasal ＞superior＞ inferior＞temporal quadrants.At the temporal,superior,nasal and inferior quadrants of inner race,the temporal quadrant of the outer race,the central area,the macular thickness and volume values were declined in the attacked eyes and fellow eyes compared with the normal control eyes (all at P＜0.05).Conclusions The edema of RNFL is more serious than that of macula in the eyes within 2 weeks after acute angleclosure glaucoma attack.The retinas of macular zone are thinner,and the macular volumes are smaller in the attacked eyes and fellow eyes than those in the normal control eyes.

Adolescent ; Anal Canal ; physiopathology ; Child ; Constipation ; physiopathology ; Female ; Gastrointestinal Motility ; physiology ; Humans ; Male ; Rectum ; physiopathology

Adolescent ; Child ; Child, Preschool ; Chronic Disease ; Colonoscopy ; methods ; Diarrhea ; diagnosis ; pathology ; Female ; Humans ; Infant ; Infant, Newborn ; Male

A rapid method was developed for the determination of 9 organophosphorous flame retardants ( OPFRs) in textiles by ultra high performance liquid chromatography tandem electrospray ionization mass spectrometry ( UHPLC-MS/MS ) . Analytes in textile samples were extracted in a closed bottle at 50℃ with methanol by ultrasonic extraction for 30 min. And then, the extracted solution was injected after filtration and determined by UHPLC-MS/MS. Target analytes were separated by C18 column and methanol/water containing 5. 0 mmol/L NH4 COOH. The positive electrospray ionization and multiple reaction monitoring mode ( MRM) was utilized. The matrix effect (ME) could be compensated by adjusting the concentration of NH4COOH and the composition of the mobile phase after careful investigation with blank samples. An external standard calibration method was chosen to the quantitative analysis. Under the optimized conditions, method precision represent with relative standard deviations (RSDs) was 5. 2% (TOCP) -12. 5% (TRIS) for cotton, 6. 4%(TBEP)-13. 8% (TRIS) for polyester, and 6. 9% (TDCP) -14. 5% (TEPA) for wool samples. The recoveries for standard addition were in the range of 52 . 5% ( TEPA ) -116 . 4% ( TPP ) . The limits of quantification (LOQ, S/N=10) were between 1. 0 (TBP) and 10. 0 (TRIS) μg/kg.

Objective To establish the quantitative detection of capsule associated protein 10 (CAP10)and virulence-associated DEAD-box RNA helicase 1(VAD1)genes in Cryptococcus neoformans (CN) and compare the diagnostic values of single gene test and combined gene test in CN meningitis.MethodsTwenty-three CN meningitis patients with fungal culture or ink staining or CN antigen detection positive in cerebrospinal fluid (CSF) were recruited and patients with craniocerebral trauma were recruited as controls.Standard plasmids were constructed using standard CN strain.Real time fluorescent quantitative polymerase chain reaction (RT-FQ-PCR) was established to detect the mRNA expressions of CAP10 and VAD1 genes in the CSF of patients with CN meningitis,which were compared with the results of CSF ink staining,fungal culture and antigen detection.The diagnostic values of single gene test and combined gene test were compared by chi square test.Results Among 23 CN meningitis patients,22 (95.6％) were CAP10 mRNA positive detected by RT-FQ-PCR,which was significantly higher than both ink staining (16/23,69.6％,x2 =4.167,P＜0.05) and fungal culture (15/23,65.2％,x2=5.143,P＜0.05),respectively; but not significant different from antigen detection (21/23,91.3％,x2=0.500,P＞0.05).There were also no statistical significant differences between combined detection of CAP 10 + VAD1 and CAP 10 or VAD1 single gene test (P＞0.05).ConclusionRT-FQ-PCR detection is successfully established using virulence genes as target,which is superior to the conventional methods.