Objective To understand clinical characteristics of primary pulmonary cryptococcosis (PPC),so as to provide reference for clinical diagnosis and treatment.Methods Clinical data of 32 patients who were confirmed PPC by pathological diagnosis in a hospital from January 2008 to June 2014 were analyzed retrospectively. Results All 32 cases were community-acquired infection ,26 male and 6 female (age between 17 and 62 years old, the average age was (35.53 ± 11 .29).Among 32 patients,8 had underlying diseases,2 were parturients,4 were carpenters,5 were pigeon keepers,3 were seafood transport drivers,and 10 lived in humid environment.Imaging findings:Solitary or multiple nodules and cluster shape(n=21),lobar consolidation(n=2),diffuse patchy shadow on bilateral lung (n=5),pulmonary cavity(n=3),and diffuse and mixed lesions (n=1).Pathological confirmation:di-agnosis through percutaneous lung biopsy(n=23),thoracoscopic surgery(n=7),and thoracotomy(n=2).Progno-sis:30 were cured,and 2 had marked effect.Conclusion PPC is commonly occurs in young and middle-aged immu-nocompetent persons,the onset is occult,clinical manifestations and imaging features lack specificity,can be easily misdiagnosed or omitted diagnosis,diagnosis is difficult,lack rapid diagnostic method in clinical practice,invasive pathological biopsy can be used as the basis of diagnosis;there is a controversy on therapy,adverse reaction of flu-conazol is mild,and has good therapeutic effect.

Genotype ; Humans ; beta-Thalassemia

Objective To investigate the correlation between the erythrocyte indices and the genotypes of alpha thalassemia.Methods337 carriers with various genotypes of alpha-thalassaemia ( iron deficiency,alpha-thalassemia double heterozygote and homozygote,α-compounding β-thalassemia and abnormal hemoglobinopathy were excluded) were classified into three groups based on different genotypes of alpha-thalassaemia including silent thalassemia group (ST,83 cases),α-thalassemia trait group (TT,210cases) and intermediate thalassemia group( IT,44 cases),and 154 healthy adults were randomly choosed as normal control The erythrocyte indices involving in RBC,Hb,MCV,MCH,MCHC and RDW-CV were retrospectively analyzed and the difference of which was compared by analysis of variance and SNK test among aboved-mentioned groups.ResultsThere were statistical significance among groups about erythrocyte indices except Hb F.The order of the level of MCV and MCH was NC [( 86.6 ± 5.2) fl,( 29.5 ± 2.1 ) pg] ＞ST[(80.1 ±3.3) fl,(26.7±1.3) pg] ＞TT[(68.5 ±3.4) fl,(22.0 ±1.2) pg] ＞IT[(66.6±7.1)fl,(20.0 ±2.2) pg,F =580.67,761.19,P ＜0.05].And the size of RDW-CV was IT(22.3 ±3.4)％ ＞TT (14.9±1.2) ％ ＞ST(13.8±1.6)％ ＞NC(13.2±1.4)％(F=347.25,P＜0.05).In ST group,the value of MCHC of -α3.7/αα subgroup( 335.6 ± 8.0) g/L was higher than that of -α4.2/αα subgroup( 330 ±7.2) g/L and αTα/αα subgroup (328.4 ±9.5) g/L(F=6.07,P ＜0.05).Meanwhile,in IT group,the value of MCV of αTα/--SEA subgroup( 70.1 ± 7.2 ) fl was higher than that of -α3.7/--SEA subgroup ( 63.4 ±5.9) fl and -α4.2/--SEA subgroup ( 64.1 ± 4.0 ) fl ( F =5.55,P ＜ 0.05 ).However,the value of MCHC of αTα/--SEA subgroup( 289.7 ± 21.2 ) g/L was lower than that of other two subgroups [( 306.3 ± 8.4 ),(306.1 ± 8.7) g/L,F =8.72,P ＜0.05].Except Hb A2 and Hb F,there was positive correlation between the number of deleted α-globin gene and that of RBC and RDW-CV ( r =0.318 and 0.580,P ＜0.01 ).Nevertheless,there was negative correlation between the number of deleted α-globin gene and that of the other erythrocyte indices (r =-0.483,-0.827,-0.744 and -0.684,P all ＜0.01 ).ConclusionsThere is close correlation between the degree of anemia and the number of deleted α-globin gene characterized by Hb reduction and RBC increasing.In addition,the anemia degree of non-deletional Hb H disease is severer than that of deletional Hb H,which of Hb H disease with -α4.2/--SEA is severer than that with -α3.7/--SEA.

ObjectiveTo evaluate the reliability of the probe melting curve analysis (PMCA) based on real-time fluorescent PCR assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.MethodsA total of 135 cases of peripheral blood samples were collected from Zhuhai Municipal Maternity and Child Healthcare Hospital between 2008 and 2010.All the samples were performed preliminary diagnosis according to the hematological data.Of these,119 cases were diagnosed as β-thalassemia trait,4 cases were diagnosed as severe thalassemia and 12 cases were normal.In addition,44 cases of amniotic fluid and 8 cases of cord blood with high-risk for severe β-thalassemia were also collected.The diagnostic reliability of the PMCA assay and reverse dot blot assay was evaluated on 187 above-mentioned cases by direct DNA sequencing analysis in a double-blind study.ResultsThe genotypes of 185 cases were detected accurately based on the PMCA assay except for two cases:one heterozygote with Ini( ATG ＞ AGG) was omitted and another heterozygote couldn't be distinguished between CD43 ( G ＞ T) and CD37 ( G ＞ A ).For the RDB assay,only one heterozygote with CD71-72 ( + T) was not detected accurately in the above-mentioned cases.Compared with the DNA sequencing analysis,the sensitivity,specificity,negative predictive value,positive predictive value and diagnostic efficiency of the PMCA assay were 98.75％,100.00％,93.10％,100.00％ and 98.93％,respectively.The corresponding value of the RDB assay were 99.38％,100.00％,96.42％,100.00％ and 99.47％,respectively.There were no significant between-group differences in the diagnostic efficiency of the two assays ( P ＞ 0.05 ).The results of prenatal diagnosis were in complete concordance with the follow up results,after the birth or induced labour of the fetuses.ConclusionsThe PMCA assay can be used as an alternative and verified method of RDB assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.

BACKGROUND: Looking for effective measures to ensure the survival of the implanted stem cells against ischemia-induced hypoxia becomes the major concern in the research of cell transplantation therapy for cerebral infarction.OBJECTIVE: To study the effects of human Persephin gene transfer on hypoxia-induced apoptosis of neural stem cells.DESTGN: A randomized controlled basic study on cells.SETTTNG: Department of Neurology, the Second Affiliated Hospital of Sun Yat-sen University.MATERTALS: This study was completed in the Lin Baixing Laboratory Center of the Second Affiliated Hospital of Sun Yat-sen University from July to December in 2006. Recombinant adenovirus pAdCMV persephin was constructed in our lab. C17.2 neural stem cells were kindly provided by Prof. Snyder, Harvard Medical University, USA. Trypsin and DMEM/F12 were purchased from Gibco Company (USA), fetal bovine serum (FBS) from Sijiqing Biological Engineering Materials Co. Ltd (Hangzhou, China); Poly-lysine from Sigma Company (USA), TUNEL assay kit and FuGENE kit from Roche Molecular Biochemicals Company (Swiss), and S-P immunohistochemical detection kit and DAB reaction kit from Mycine Biological Engineering Company (Fujian). Rat anti-human monoclonal Nestin antibody and rabbit anti-human polyclonal persephin antibody were manufactured by Santa Cruz Company (USA), and persephin anti-senseoligodeoxynucleotide (ODN) was synthesized by Shanghai Biological Engineering Company.METHODS: ① Interventions: C17.2 neural stem cells cultured in vitro were infected by recombinant adenovirus containing persephin gene, and they were divided into four groups: blank control group (Group A, in which the C17.2 neural stem cells were not treated with hypoxia), hypoxic group [Group B, in which the cells were cultured at 37 ℃ in anaerobic incubation containing N2 (0.95 in volume fraction) and CO2 (0.05 in volume fraction)], hypoxia + pAdCMV persephin infection group [Group C, where the cells were cultured under the conditions as in group B after pAdCMV persephin infection for 48 hours], and hypoxia + pAdCMV persephin infection + anti-sense persephin ODN group (Group D, where the cells were infected by pAdCMV persephin and anti-sense persephin ODN. ② Evaluation: The expression of Persephin protein was analyzed using Western blotting; Apoptotic index was detected with terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end-labeling (TUNEL) assay; The changes of apoptotic rate was determined with flow cytometry.MAIN OUTCOME MEASURES: Expression of Persephin protein; Apoptotic index; Apoptotic rate.RESULTS: ① Expression of Persephin protein: A specific band (relative molecular mass of 24 000) was detected by Western blotting in pAdCMV persephin infected cells, suggesting the successful expression of persephin gene.Interestingly, the cells infected with both pAdCMV persephin and anti-sense persephin ODN also showed the specific band of about 24 000, but with much less density, indicating that anti-sense persephin ODN could effectively inhibit the expression of pAdCMV persephin. However, this band was not presented in the blank control groups. ② Apoptotic index:The apoptotic index in group C was significantly lower than those in groups B and D (P＜0.01), but still higher than that of group A (P＜0.01), suggesting that persephin gene transfer could attenuate apoptosis to some extent. ③ Apoptotic rate: The apoptotic rate in groups B and D were obviously higher than that in group A (P ＜ 0.01), and it was lower in group C than in groups B and D (P＜0.01).CONCLUSION:Recombinant adenovirus can efficiently mediate Persephin gene transfer into C17.2 neural stem cells,resulting in high expression of the exogenous Persephin in vitro, which effectively reduces C17.2 neural stem cell apoptosis induced by hypoxia.

BACKGROUND:Adipose-derived stem cels are a kind of mesenchyam stem cels with multipotent differentiation capacity, which have more advantages than bone marrow mesenchymal stem cels in tissue engineering research. OBJECTIVE: To establish a method to isolate and purify adipose-derived stem cels from human subcutaneous adipose tissues folowed byin vitro amplification and osteoblastic/adipogenic differentiation.
METHODS: Adipose-derived stem cels were isolated from human subcutaneous adipose tissue and cultured by density gradient centrifugation and adherent culture. Cel morphology and growth features were observed under inverted microscope. Adipose-derived stem cels at passages 2 and 5 were selected for viability measurement using cel counting kit-8 method, and then cel growth curves were drawn. The immunophenotype identification was analyzed by flow cytometry. Passage 5 cels underwent osteoblastic/adipogenic induction to confirm the multi-differentiation potential.
RESULTS AND CONCLUSION: (1) Using density gradient centrifugation and adherent culture method, high-purity human adipose-derived stem cels can be successfuly isolated from human adipose tissues. (2) The growth process of human adipose-derived stem cels includes stagnant phase, logarithmic phase and plateau phase, which meets the growth rhythm of normal cels. Moreover, the population doubling time is shorter. (3). Human adipose-derived stem cels are positive for stem cel-related antigens, with low immunogenicity and the multi-differentiation potential. (4) Labeling human adipose-derived stem cels with DAPI is a simple efficient labeled method, and the labeling rate is high but the cytotoxicity is low

Objective To study the therapeutic effect of adjuvant chemotherapy and radiotherapy after operation in patients with mucoepidermoid carcinoma of parotid gland,and to screen the indicators ralated to the prognosis of tumor.Methods Eighty patients with mucoepidermoid carcinoma of parotid gland in First People′Hospital of Yibin of Sichuan Province from January 2005 to December 2009 were analysed retrospectively in our research.We studied the survival of patients who were treated wtih simple operation(30 cases)or postoperative adjuvant therapy(50 cases).Then we further analyzed the relationships between the prognosis of the patients and some variables (age,gender,smoking,alcohol drinking,lymph node metastasis,distant organ metastasis,treat-ment method,differentiation degree and T grading).Results Kaplan-Meier survival curves showed that patients with postoperative adjuvant therapy had longer PFS and OS than those without adjuvant therapy (94.4 months vs 69.3 months;114.9 months vs 96.7 months),with statistical significance (χ2 =11 .246,P =0.001 ;χ2 =15.803,P =0.001 ).COX univariate analysis showed that gender (χ2 =22.346,P =0.000),smoking (χ2 =7.891 ,P =0.041 ),lymph node metastasis (χ2 =12.371 ,P =0.005),distant organ metastasis (χ2 =9.81 3, P =0.002),treatment method (χ2 =25.261 ,P =0.000),differentiation degree (χ2 =4.361 ,P =0.006)and T grading (χ2 =5.336,P =0.01 4)were related to the PFS of patients.COX multivariate analysis showed that lymph node metastasis (χ2 =11 .003,RR =2.827,95%CI:1 .965-3.851 ,P =0.011 ),distant organ metastasis (χ2 =7.611 ,RR =0.472,95%CI:0.240-0.775,P =0.016),treatment method (χ2 =24.542,RR =5.390, 95%CI:3.585-9.602,P =0.000),degree of differentiation (χ2 =3.221 ,RR =2.1 1 8,95%CI:1 .845-4.719, P =0.009)and T grading (χ2 =4.336,RR =0.804,95%CI:0.681 -0.916,P =0.024)were related to the PFS of patients.COX univariate analysis showed that smoking (χ2 =4.551 ,P =0.008),alcohol drinking (χ2 =11 .742,P =0.048),lymph node metastasis (χ2 =14.886,P =0.009),distant organ metastasis (χ2 =6.71 3, P =0.005),treatment method (χ2 =22.411 ,P =0.000),degree of differentiation (χ2 =8.1 16,P =0.012)and T grading (χ2 =14.443,P =0.035)were related to the OS of patients.COX multivariate analysis showed that lymph node metastasis (χ2 =11 .711 ,RR =2.985,95%CI:1 .521 -3.999,P =0.005),distant organ metastasis (χ2 =5.390,RR =0.400,95%CI:0.201 -0.793,P =0.009),treatment method (χ2 =19.327,RR =5.086, 95%CI:3.241 -8.006,P =0.000),degree of differentiation (χ2 =7.084,RR =2.301 ,95%CI:1 .908-4.503, P =0.001 )and T grading (χ2 =1 3.229,RR =0.561 ,95%CI:0.348-0.867,P =0.040)were related to the OS of patients.Conclusion Adjuvant radiation and chemotherapy can obviously prolong the PFS and OS for the patients with mucoepidermoid carcinoma of parotid gland.Lymph node metastasis,distant organ metastasis,treat-ment method,differentiation degree and T grading can greatly influence the prognosis of patients with mucoepider-moid carcinoma of parotid gland,which can be used as independent prognostic indicators for the patients with mucoepidermoid carcinoma of parotid gland.

Purpose To investigate the diagnostic value of high-resolution CT reconstruction techniques on the same slice in hypertrophy of transverse process of the fifth lumbar vertebra (HTPL5V), and to provide a basis for clinical diagnosis and treatment. Materials and Methods Twenty-two cases of clinically diagnosed HTPL5V and 20 normal adults were examined with GE LightSpeed 16-slice spiral CT (36 cases) and Philips iCT 256-slice (6 cases). L5 transverse process and the fifth lumber nerve were reconstructed and observed on the workstations. Results In 22 cases of HTPL5V, there were 26 pseudarthrosis formation and 2 sides with L5 transverse process touching the sacral ala. In 28 sides the iffth lumber nerve traveled through false foramina of the HTPL5V including 6 cases of bilateral compression and 16 cases of unilateral compression. In 21 cases, the nerve was compressed by hyperosteogeny on 27 sides (96.4%) and 1 side due to stenosis (3.6%). On 25 sides (89.3%) the compressed nerves were curved in shaper. There was bulging and/or herniated lumbar disc on 9 sides in 7 cases (32.1%). Conclusion High-resolution CT reconstruction techniques can demonstrate the iffth lumbar nerve of HTPL5V and provide evidence for clinical diagnosis and treatment.

Objective To evaluate the curative effect of sedation on broncholiths through bronchoscope with holmi-um laser. Methods From Jan.2008 to Dec. 2015, 12 cases with broncholiths through bronchoscope with holmium laser, male 7, female 5; the age from 35 to 64.12 cases visit a doctor when they cough, panting, haemoptysis and cough out stones. The predilection sites of bronchinal calculus: left main bronchus in 6, right middle bronchus in 4, main bronchus in 2. Thoracic computerized tomography was performed in 12 patients, which showed bronchial intra-luminal high-density shadow with distal bronchial stenosis, bronchiectasis, COPD, hilar calcifications, or mediastinal lymph node calcifications. Through bronchoscopy examination was performed in 12 cases, Broncholiths were found in 9 patients and granulomatous lesion wrapping hard lesions in 3 patients.12 cases were treated by using Dexmedeto-midine combined with Sufentanil for sedation through of bronchoscope with holmiun laser. Results All the operations were successful, the operation time 45~90 min, average 60 min . During the surgery, the patient have stable heart rate, oxygen saturation without falling, blood pressure is stable, stable hemodynamics, the patients did not complain of discomfort, without obvious heart and lung failure and other complications occurred, the operation without bleed-ing, pneumothorax, complications. All cases of postoperative respiratory system were improved after operation. The average time stay in hospital was (2.5 ± 1.4) days. Follow-up for 1~24 months (mean, 6 months) in 12 cases found no recurrence of stones and serious respiratory tract infection. Conclusions The method of Dexmedetomidine com-bined with Sufentanil for sedation on broncholiths through of bronchoscope with holmiun laser is safey, feasible and therapeutic effects were clearly, which provides a new method for the clinical on Broncholiths.

Horizontal gene transfer (HGT), also Lateral gene transfer (LGT), is any process in which an organism transfers genetic material to another species that is not its offspring. With the increase of available genomic data, it has become more convenient to study the way to detect the genes, which are products of horizontal transfers among a given genome. There are few data about known horizontal gene transfers in three bacterium genomes under consideration, so the experiments, which simulated gene transfer by artificially inserting phage genes, were carried out. Combining the feature analysis methods of gene sequences with support vector machine (SVM), a novel method was developed for identifying horizontal gene transfers (HGT) in 3 fully sequenced bacterium genomes (Escherichia coli K12, Borrelia burgdorferi, Bacillus cereus ZK). According to our previous work, codon use frequency (FCU) was selected as the sequence feature, in respect that it is inherently the fusion of both codon usage bias and amino acid composition signals. In addition, another computational method was proposed considering strand asymmetry and predicting horizontal gene transfers of leading strand and lagging strand of genomes under consideration, respectively. To avoid the occasionality of simulating gene transfer through artificially inserting phage genes, 100 times of the transfer-and-recover experiment were repeated and arithmetic average of measurement for each genome being considered were reported to evaluate algorithm's performance. Ten-fold cross-validation was used for both parameter and accuracy estimation. The best results were obtained for C-Support Vector Classification (C-SVC) type by using the radial basis function kernel with ?=100, while for one-class SVM type the best performance was obtained using the polynomial kernel of three degree. The performance of the approach was compared with that of Tsirigos' method ,which is one of the best predictive approachs to date in detecting of horizontal transfer genes. Firstly, for the original method that did not consider the strand asymmetry, the C-SVC type has a high relative improvement(RI) of 31.47% on hit ratio for Escherichia coli K12, while the one-class SVM type has RI of 11.61% for Borrelia burgdorferi. Moreover, as theoretically expected, the method considering the strand asymmetry resulted in higher RI than the original method. In order to examine the approach's performance in detecting factual gene transfer events, the approach was applied in genome of Enterococcus faecalis V583. It is not only succeed in recovering all the seven factual horizontally transferred genes, also found that the whole segment from 7 kb upstream of gene EF2293 to 38 kb downstream of gene EF2299 was probably transferred into E. faecalis V583 genome simultaneously with the above seven genes.