To screen specific DNA probes by double hybridization from the constructed DNA li- brary of serotype A Cryptococcus neoformans.On the basis of the nucleotide sequence of vector pUC18, a pair of primers was synthesized.The insert fragments were amplified from the library on a PCR pro- cessor.The PCR products were spotted onto hybond-N~+ membranes.All inserts amplified from the ge- nomic library by PCR were screened by dot blot with 26 ~(32)P-labelled DNA from infectious agents for dif- ferentiation and from healthy persons.Twenty-eight candidate clones were obtained.The twenty-eight clone inserts got from low melting point agar were further characterized by dot blot with above 27 kinds of DNA for differentiation.Three specific DNA probes from the library of serotype A Cryptococcus neo- formans were obtained.Colony pCNIIA6 was serotype A-specific,which gave signals only with sertype A strain and did not cross hybridize with other DNAs.Colony pCNIIB5 was species-specific which gave signals only with DNA from Cryptococcus neoformans.Colony pCNIIIG1 was specific for var.neofor- mans,which gave signals only with serotype A and D strains.We can make a rapid diagnosis of Crypto- coccus neoformans infection at an early stage and distinguish the variants of C.neoformans and C.gattii using above specific probes.

Polymerase chain reaction(PCR) was used for the first time in China for the diagnosis of cryptococcal meningitis by amplification of specific sequence of the rDNA genes of C.neoformans.All 11 strains of C.neoformans yielded a specific 136 bp fragment but 21 non-C,neoformans strains did not. The sensitivity of the amplification was about 10 cfu.With the aid of the rDNA-PCR,23 of the 23 cere- brospinal fluids (CSF) specimens which had been confirmed C.neoforrnans positive by smear and/or cul- ture were PCR-positive(100%),and 13 of the 14 CSF specimens which had been confirmed C.neofor- mans negative by smear and culture were PCR-negative (93%).The results of the present study suggest that rDNA-PCR is a sensitive and specific method for rapid diagnosis of cryptococcal meningitis.

Objective To observe the curative effect of Qujun Tongyou Decoction instead of antibiotics in treatment of Helicobacter pylori (HP) associated gastric disease, and explore the mechanism of action. Methods In 236 patients, 140 cases of newly diagnosed patients were randomly divided into treatment group of 80 cases and control group of 60 cases, 96 cases of retreatment patients were randomly divided into treatment group of 56 cases and control group of 40 cases. The control group was given western quadruple therapy:newly diagnosed patients with rabeprazole sodium enteric-coated capsules+colloidal bismuth pectin+Amoxicillin Capsules+clarithromycin dispersible tablets;retreatment patients with rabeprazole sodium enteric-coated capsules+colloidal bismuth pectin+metronidazole tablets+levofloxacin tablets. The treatment group was given rabeprazole sodium enteric coated capsules and colloidal bismuth pectin plus Qujun Tongyou Decoction. All patients were treated for 2 weeks. 1, 3, 6 months after treatment, 14C-urea breath test was detected, HP negative rate, recurrence rate, clinical symptoms, and adverse reaction were observed. Results HP negative rate, the total effective rate and the recurrence rate in treatment group were different with the control group (P<0.05). There was significant difference in the rate of symptom control between treatment group and the control group (P<0.01). No adverse reaction showed in treatment group. Conclusion Rabeprazole sodium enteric-coated capsules and colloidal bismuth pectin combined with Qujun Tongyou Decotion can effectively inhibit and kill HP, promote the repair and healing of ulcer, control symptoms, reduce HP recurrence, lessen adverse reaction, is effective in the treatment of HP associated gastric diseases.

Objective: To evaluate the influence of acute moderate hemodilution on the dose-response and timecourse of effect of vecuronium. Method:Sixty patients. ASA grade Ⅰ,aged 17-45 years,scheduled for elective plastic surgery were included in the study,of which,thirty patients underwent hemodilution during surgery and thirty patients did not receive hemodilution as controls. General anesthesia was maintained with 60% nitrous oxide in oxygen,and further increments of thiopental 2 mg/kg or fentanyl 2?g/kg as required. After anesthesia was stable,the status of acute moderate hemodilution was developed by drainage o{ venous blood and intravenous infusion of lactated Ringer'ssolution, 6% dextran and.gelofusine,during which the level of hemoatocrit dropped from 45.7% to 26.2%. Neuromuscular function was assessed by TOF stimulation of accelerometry with the percentage depression of T_1 response used as the study parameter. The dose-response relationships of vecuronium were determined with the cumulative dose-response technique. Result:During hemodilution the dose-response curve of vecuronium was parallelly shifted to the left. Compared to the control patients,ED_(50),ED_(90) and ED_(95) of vecuronium in the hemodilution patients were decreased by 22%,18% and 17%, respectively. Following an intravenous administration of total dose of vecuronium 80?g/kg, vecuronium-induced neuromuscular block was significantly longer in the hemodilution patients than in the control patients. Conclusion:Acute moderate hemodilution could significantly enhance the neuromuscular blocking effect of vecuronium and prolong its duration of action.

Objective To evaluate the effects of different treatments to middle turbinate by endo-scopic sinus surgery (ESS) on olfaction in patients with chronic sinusitis and nasal polyps. Methods ESS was performed on 74 cases of chronic sinusitis and nasal polyps. Proper treatments divided into the group of retaining middle turbinate, the group of forming middle turbinate and the group of partial endonasal middle turbinectomy which were applied to different pathological changes of middle turbinates. The olfactory function of the 74 patients was tested through the CCCRC olfactory testing method pre-operation and twenty-four weeks post-operation. The results were analyzed by analysis of variance. Results The olfaction after treat-ment of the three groups was significantly better than that of pre-operation (P < 0.05), but there was no sig-nificant difference among the three groups (P>0.05). Conclusion Different treatments to middle turbinate by ESS is no obvious effects on olfaction.

Objective To analyze the application of recombinant gp125 proteins in serological diagnosis of nasopharyngeal carcinoma (NPC). Methods B cell epitopes of EBV gp125 protein were forecasted by internet server and the peptides were synthesized and coated to microplate for the detection of EBV-IgA antibody in NPC patients and healthy donors by ELISA. Results Six peptides were synthesized, wherein two showed low specificity and others displayed significantly higher positive rate in the NPC group than in the healthy group (P <0.001). Conclusion The BALF4 gene peptides are successfully synthesized and epitopes with high specificity and strong immunogenicity are screened for the preparation of gp125 antigen with high specificity and sensitivity.

Objective To screen and identify the novel genes from the adult worm cDNA library of Schistosoma japonicum by the expressed sequence tags (EST) strategy.Methods The cDNA clones were selected randomly and the ESTs were obtained.The novel gene was searched for homologue identity with NCBI blast programmer.The homologue of the two sequences with a high identity was compared at amino acid and nucleotide level with the BLAST programmer,and the analysis of protein was carried out with the pcgene software. Results There was a novel gene of Schistosoma japonicum which included 1 677 bp coding for 424 amino acid residues, and it was given the accession number of sequence in Genbank(AY336497).The cDNA sequence was homologous to the known ornithine aminotransferase gene of Xenopus laevis. The identity of amino acid sequence was 66%. The academic pI was 8.52, and the antigen determinant was probably from 795 to 846 on the cDNA sequence. Conclusion EST strategy is an effective measure to discover new genes of Schistosoma japonicum. The novel gene is homologous to the ornithine aminotransferase gene of Xenopus laevis.This study is helpful to further sequential and functional search of the gene.[

Objective To relate the results of surface electromyography (sEMG) of the femoris among patients with hemiplegia after a brain lesion with scores on the modified Ashworth scale (MAS),and to decide whether sEMG can be used as a quantitative index for evaluating spasm.Methods The affected quadriceps femoris of sixty hemiplegic stroke survivors were assessed by the same therapist using the MAS.sEMGs were also recorded and the root mean square and average sEMG readings from the vastus medialis (VM),rectus femoris (RF) and vastus lateralis (VL) were extracted.The correlation between these sEMG indexes and the MAS grading was analyzed.Results The average sEMG signals from the VM,RF and VL were strongly correlated with the MAS grading,and they increased with increasing MAS grade.Spearman correlation analysis demonstrated highly significant correlation (P ≤ 0.001) when the quadriceps femoris was moved passively.Conclusions Surface EMG signals from the VM,RF and VL correlate well with MAS grades,rising when the affected quadriceps femoris of a stroke survivor is moved passively.sEMG can present the electrical activity during passive movements of the quadriceps femoris and can be useful in spasticity assessment and clinical treatment.

Objective To investigate the effect of CYP3A5 and C5 gene polymorphisms on the concentration/dose ratio of tacrolimus in Chinese liver transplant patients during the early posttransplantation period.Methods A total of 100 adult patients who underwent primary liver transplantation (LT) were enrolled.Tacrolimus dosage and trough blood concentration were detemined at first week after liver transplantation.Two single-nucleotide polymorphisms were genotyped and analyzed in both donor and recipient groups.The relationship between gene polymorphisms and tacrolimus concentration/dose ratio (C/ D ratio) was analyzed.Results The distribution of allele A in C5 rs17611 was 63.5 ％ among donors and 58.5％ among recipients.For CYP3A5,the rs776746 allele G represented the major alleles in both donors and recipients (71％ and 72％,respectively).The tacrolimus C/D ratio of recipients carrying allele AA in C5 rs17611 was significantly higher than that of recipients carrying the C5 rs17611 allele G.Both donor and recipient CYP3A5 rs776746 polymorphisms were highly correlated with the tacrolimus C/D ratio at first week after liver transplantation.No linkage disequilibrium between CYP3A5 rs776746 and C5 rs17611 polymorphisms was found (D'max =0.392,r2max =0.034).Recipient CYP3A5 rs776746 allele A,C5 rs17611 genotype AA,and donor CYP3A5 rs776746 allele A were associated with rapid tacrolimus metabolism.With increasing number of these alldes,tacrolimus C/D ratio was reduced during the one week after transplantation.Conclusion Recipient C5 rs17611 polymorphism is a new genetic locus that influences tacrolimus metabolism in patients after OLT during the early post-transplantation periocd.

Objective To investigate the clinical features of urothelial carcinoma in a single center.Methods The clinical data of 2 115 urothelial carcinoma patients admitted to Peking Union Medical College Hospital from January 2003 to December 2012 were reviewed retrospectively.Of the 2 115 patients,1414 cases were bladder carcinoma,329 cases were ureter carcinoma and 372 cases were renal pelvic carcinoma.Statistical method was used to analyze the differences of distribution and pathological grading and staging of bladder,ureter and renal pelvic carcinoma among different ages,genders,and urban and rural patients.Results The proportion of low-grade carcinoma was 83.5％ for bladder,68.7％ for ureter and 46.5％ for renal pelvic carcinoma,and the proportion of non-muscle invasive carcinoma was 84.9％,74.8％ and 59.7％ respectively,and significant difference was found in pathological grading and staging among the three groups (P =0.000).In bladder and renal pelvic carcinoma patients,males were common,while females were more common in ureter carcinoma patients.There was significant difference in gender among the three groups (P =0.000),while there was no significant difference in pathological grading and staging between male and female patients(P＞0.05).89.7％ (1 898 cases) were concentrated in the 41-80 years old.The grading and staging of urothelial carcinoma in young,middle-aged and elderly groups showed significant difference (P＜ 0.05).The proportions of high-grade and muscle invasive urothelial carcinoma patients in rural areas were both higher than that in urban areas,the difference in pathological grading and staging between urban and rural patients was significant (P＜0.05).Of the 129 cases (6.1％) with multifocal urothelial carcinoma,there were 89 cases (70.0％) with high grade and 62 cases (48.1％) with muscle invasive carcinoma.Significant difference was found in pathological grading and staging between multifocal and single urothelial carcinoma (P=0.000).Conclusions In the uretholial carcinoma of Peking Union Medical College Hospital in the past ten years,the proportion of bladder carcinoma is the highest,while ureter and renal pelvic carcinoma have higher proportion of high grade and stage.The proportion of patients in the 41-80 years old is higher.The pathological grading and staging of urothelial carcinoma is positively correlated with age,but there is no significant correlation in gender.The patients with high grade and stage carcinoma in rural areas are more than that in urban areas.Multifocal urothelial carcinoma has higher malignancy.