Genotype ; Humans ; beta-Thalassemia

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.

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.

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.

Objective To evaluate the effects of stepwise and fine-grained dietary management on the diet control of liver cirrhosis patients complicated with gastrointestinal hemorrhage. Methods Eighty liver cirrhosis patients complicated with gastrointestinal hemorrhage were assigned to experimental group (41 cases) and control group(39 cases) by random digits table method. The patients in experimental group were treated with stepwise and fine-grained dietary management and the patients in control group were given conventional dietary treatments. The compliance diet, mastery of nutrition knowledge, malnutrition risk at admission, intake food, discharge and 2 months after discharge between two groups were monitored. The incidence of rebleeding was tracked after 1 year of discharge. Results The incidence of malnutrition risk in Child-Pugh class C was higher than that in Child-Pugh class B( Z=-4.3, P<0.05 ) . The incidence of malnutrition risk in patients with high education level was lower than that in patients with lower education( r=-0.453, P<0.05 ). The experimental group significantly outperformed control group in mastery of nutrition knowledge at discharge:80(70, 84) points vs. 52(42, 64) points, in compliance diet and malnutrition risk after 2 months after discharge:86(76, 91)%vs. 53(46,57)%, 1(1, 2) points vs. 2(1,3) points, the rebleeding rate in experimental group was significantly less than that in control group within 1 year after discharge 14.6%(6/41) vs. 35.9%(14/39), and the differences were statistically significant between two groups ( χ2 = 4.226- 51.232, all P < 0.05 ). Conclusions Stepwise and fine-grained dietary management can improve the mastery of nutrition knowledge, compliance diet and nutritional status, and reduce the occurrence of rebleeding, worth popularizing further in clinical care of liver cirrhosis patients complicated with gastrointestinal hemorrhage.

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.

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 investigate the clinical value of the probe melting curve analysis‐based PCR assay for the detection of three types of αT .Methods A total of 149 blood and prenatal archival DNA samples (6 of which were amniotic fluid samples)with three knownαT genes ,which included 63 carriers with Hb CS ,22 cases with Hb QS ,43 individuals with Hb WS and 1 double heter‐ozygote with Hb CS and Hb WS) as well as 20 samples with normalα‐globin gene sequence that had been confirmed by RBD com‐bined with DNA sequencing were selected to test the specificity of probe melting curve analysis by blind analysis .Results The probe melting curve analysis accurately detected 100 of the DNA samples previously characterized by S RBD combined with DNA sequencing .Conclusion Probe melting curve analysis‐based PCR assay for the detection ofαT is featured with rapidity and accuracy and can be applied to clinical and prenatal diagnosis .

BACKGROUND: Changes of elastic fibers in middle cerebral artery(MCA)is close related withthe aged cerebrovascular disease.OBJECTIVE: To observe the changes of elastic fibers of MCA in different aging rats.DESIGN: A descriptive and controlled study based on experimental animals.SETTING: Department of anatomy and central laboratory in a university.MATERIALS: Totally 36 healthy Wistar rats with either gender, weighing 200 - 280 g, were selected from the Animal Institute of the third medical military university of Chongqing[certification SCXX (army) 2002-007].INTERVENTIONS: Changes of elastic fibers of MCA of different aging rats were observed with light microscope, transmission electron microscope and image analysis system. MAIN OUTCOME MEASURES: ①) Major outcome: changes of elastic lamella in MCA of different aging rats; ②) Secondary outcome: ultramicrostructural changes of internal lamella under the transmission electron microscope. RESULTS: With the increase of age, the folded extent and quantity of internal elastic lamella were decreased, and the content of elastic fibers were also decreased significantly( P ＜ 0.01 ). However, the ratio of collagen fibers to elastic fibers was increased significantly( P ＜ 0.01 ) . In the aging group above 24 months, the internal elastic lamina thinned, delaminated and disrupted, and the lipid deposited in it. Endothelial cells and smooth muscle cells passed through the internal elastic lamina. CONCLUSION: Changes of elastic fibers may be related with the increased susceptibility to the cerebrovascular disease in aged people.