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.

Asthma ; diagnosis ; Child ; Humans ; Perception

Disaster Planning ; Humans ; Rescue Work

The aerolysin genes (aerA) of BZ and NK isolates were cloned and sequenced. The sequence analysis showed that the partial aerA of BZ and NK isolates consisted of 1393 bp, encoding a protein of 464 amino acids. The similarity of nucleotide and amino acid sequences of aerA between BZ and NK isolates was 97.6% and 98.3% respectively. The nucleotide sequence of aerA of BZ strain exhibited 71.6% to 97.5% homology with other Aeromonas isolates, and the amino acid sequence exhibited 68.0% to 98.9% homology. The phylogenetic tree based on aerA nucleotide sequences from Aeromonas isolates was constructed with neighbor-joining method. It showed that there were three branches of aerolysin genes, and a close relation- ship among Aeromonas hydrophila isolates which were clustered into the same branch.

Objective: To observe the role of rmh-TNF in enhancing the anti-angiogenesis effect of cisplatin on Lewis lung carcinoma in the mice.Methods: Lewis lung carcinoma model was established in C57BL/6 mice.Sixty model mice were randomly divided into 4 groups: control group,rmh-TNF group(1500000 U/kg),cisplatin group(6.15 mg/kg), and rmh-TNF plus cisplatin group.Twelve days after implantation of cancer cells,different drugs were injected intra- tumorallv for 3d.The expression of hypoxia inducible factor-1?(HIF-1?)gene in the tumor was identified by RT-PCR. Immunohistochemistry(IHC)image analysis was performed to determine the vascular endothelial growth factor(VEGF) and kinase domain region receptor(KDR)expression and the microvessel density(MVD).Expression of matrix metallo- proteinase-2(MMP-2)was detected by flow cytometry.Results: The MVD values in the control group,the rmh-TNF group,the DDP group and the combination group were(24.76?1.28),(18.95?1.22),(19.53?1.15),(10.43?1.05),respectively,with those of the rmh-TNF and DDP groups significantly lower than that of the control group and higher than that of the combination group(all P

Objective Establish an overall performance evaluation index system for innovative medical devices transforming project,a major science and technology project in Zhejiang province.Methods The weight of indexes was calculated using Delphi method,analytic hierarchy process and weight of percentile method.Results A three-level performance evaluation index system was established.First level included 3 indexes,as condition index,process index and result index,with weight of 0.3401、0.4042 and 0.2557 respectively.Meanwhile,second level and third level included 13 and 38 indexes.Conclusions The index system provide a performance evaluation tool for the innovative medical devices transforming project.

The culture of Magnetospirillum magneticum WM-1 depends on several control factors that have great effect on the magnetic cells concentration. Investigation into the optimal culture conditions needs a large number of experiments. So it is desirable to minimize the number of experiments and maximize the information gained from them. The orthogonal design of experiments and mathematical statistical method are considered as effective methods to optimize the culture condition of magnetotactic bacteria WM-1 for high magnetic cells concentration. The effects of the four factors, such as pH value of medium, oxygen concentration of gas phase in the serum bottle, C:C (mtartaric acid: msuccinic acid) ratio and NaNO3 concentration, are simultaneously investigated by only sixteen experiments through the orthogonal design L16(44) method. The optimal culture condition is obtained. At the optimal culture condition (pH 7.0, a oxygen concentration 4.0%, C: C (mtartaric acid: msuccinic acid) ratio 1:2 and NaNO3 100 mg l-1), the magnetic cells concentration is promoted to 6.5×107 cells ml-1, approximately 8.3% higher than that under the initial conditions. The pH value of medium is very important factor for magnetic cells concentration. It can be proved that the orthogonal design of experiment is of 90% confidence. The results from the hysteresis of WM-1 shows that Hc = 230 Oe, Ms = 0.9 emu/g dry wt. Cells,and Mr / Ms = 0.50.

Anal Canal/surgery* ; Anorectal Malformations ; Constriction ; Fecal Incontinence ; Humans ; Rectal Prolapse

OBJECTIVETo evaluate the diagnostic specificity of dynamic assessment and monitoring using a portable spirometer in diagnosis and differential diagnosis of asthma.

METHODSWe retrospectively reviewed the results of dynamic monitoring of spirometry in 145 symptomatic patients with physician-diagnosed asthma. Flow-volume curve and simultaneous symptoms and mood were measured in a fixed-time thrice-daily assessment schedule for 2 weeks. Patients were allowed to make additional measurements in case of symptom exacerbations.

RESULTSThe clinical data of 51 males and 94 females with a mean age of (39.1 +/- 13.0) years (ranged from 10 to 65 years) were analyzed. Duration of asthma before study was (6.7 +/- 9.9) years. Of 145 patients with physician-diagnosed asthma, 126 (87%) could be conclusively confirmed for a diagnosis of asthma. Asthma was misdiagnosed in 14 patients (9.7%). Overdiagnosis of asthma was observed in 5 patients (3.4%).

CONCLUSIONDynamic assessment and monitoring using a portable spirometer by revealing variability and reversibility of airway obstruction may provide an additional tool for diagnosis and differential diagnosis of asthma.

Adolescent ; Adult ; Aged ; Airway Obstruction ; Asthma ; diagnosis ; Child ; Diagnosis, Differential ; Diagnostic Errors ; Female ; Humans ; Male ; Middle Aged ; Peak Expiratory Flow Rate ; Spirometry ; methods ; Young Adult

Objective: To investigate the effect of rhubarb on gastrointestinal blood perfusion in critical illness and hemorrhagic shocked rats.Methods: Clinical Study: Sixty-four septic patients, who suffered from stress ulcer, were treated with rhubarb at a dose of 25 mg/kg. Twenty-five non-septic patients were taken as control. The gastrointestinal perfusion was evaluated by intramural pH (pHi). Animal study: SD rats were anesthetized with intraperitoneal sodium pentobarbital at a dose of 20 mg/kg. Blood-letting were performed in the animals. Blood pressure reduced to 5.32 kPa and maintained for 120 mins. They were resuscitated at the end of shock by reinfusing all of the shed blood. The rats were randomly divided into four groups: Normal control, shock group, therapeutic group (shocked rats were treated with 50 mg/kg rhubarb at the end of shock) and rhubarb group (normal rats were treated with rhubarb). Laser Doppler was applied to estimate the gastrointestinal blood perfusion. Results: Clinical Study: The gastrointestinal pHi in septic patients was much lower than that in the control, whereas rhubarb could obviously elevate gastrointestinal pHi (P<0.001). In addition, rhubarb also had good effect on gastric hemorrhage caused by stress ulcer. Animal Study: Although the shocked rats were resuscitated completely, their gastrointestinal blood perfusion was much lower than that in the control. Rhubarb could significantly improve the blood perfusion in gastrointestinal mucosa and mesentery (P<0.01). Furthermore, rhubarb also increase the gastrointestinal perfusion in normal rats. Conclusion: Rhubarb could improve gastrointestinal blood perfusion in critical illness and shocked rats.