Lentiviral vectors have drawn considerable attention recently and show great promise to become important delivery vehicles for future gene transfer manipulation. In the present study we have optimized a protocol for preparation of human immunodeficiency virus type-1 (HIV-1)-based defective lentiviral vectors (DLV) and characterized these vectors in terms of their transduction of different cells. Transient co-transfection of 293T packaging cells with DNA plasmids encoding lentiviral vector constituents resulted in production of high-titer DLV (0.5-1.2 × 107IU/mL), which can be further concentrated over 100-fold through a single step ultracentrifugation. These vectors were capable of transducing a variety of cells from both primate and non-primate sources and high transduction efficiency was achieved using concentrated vectors. Assessment of potential generation of RCV revealed no detection of infection by infectious particles in DLV-transduced CEM, SupT-1 and MT-2 cells. Long-term culture of transduced cells showed a stable expression of transgenes without apparent alteration in cellular morphology and growth kinetics. Vector mobilization to untransduced cells mediated by wild-type HIV-1 infection was confirmed in this test. Challenge of transduced human T-lymphocytes with wild-type HIV-1 showed these cells are totally resistant to the viral infection. Considering the effective gene transfer and stable gene expression, safety and anti-HIV activity, these DLV vectors warrant further exploration for their potential use as a gene transfer vehicle in the development of gene therapy protocols.

OBJECTIVETo explore the essence of Spleen-Kidney Deficiency in middle-aged patients.

METHODSInvestigation was carried out in 773 cases of 50-69 years old to assay 34 parameters in them, including blood lipid, oxidation and anti-oxidation related substance, sex hormone, liver and renal function, immune function, blood routine, blood pressure and lung vital capacity, etc.

RESULTSSpleen Deficiency Syndrome is closely related with lipid metabolism disorder; Kidney-Qi Deficiency Syndrome is closely related with hypoimmune function; Kidney-Yin Deficiency Syndrome is closely related with lipid metabolism disorder and hypertension, and Kidney Yang Deficiency is closely related with weakness of anti-oxidation capacity, hypoimmunity, the internal environment disorder of sex hormone and aging, manifested as multiple functions abating.

CONCLUSIONSpleen Deficiency, Kidney-Qi Deficiency and Kidney Yang Deficiency are different layers of a gradually developed and aggravated pathological process, but Kidney-Yin Deficiency could not be listed into this layer.

Aged ; Aging ; CD4-CD8 Ratio ; Cholesterol ; blood ; Diagnosis, Differential ; Female ; Humans ; Kidney Diseases ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Splenic Diseases ; blood ; Superoxide Dismutase ; blood ; Yang Deficiency ; blood ; Yin Deficiency ; blood

Objective To probe the oprⅠ gene in rat model with Pseudomonas aeruginosa septicemia by FQ-PCR,and compare the sensitivity and specificity between FQ-PCR and traditional germiculture,and check the change of oprI gene before and after the antibiotic therapy as to rapidly judge its sensitivity.Methods The standard Pseudomonas aeruginosa with five different concentration were prepared,the drug-sensitive test wbre used to find lhe sensitive antibiotics.120 SD rats were random divided into five groups,five different concentrations of Pseudomonas aeruginosa were injecked into the rats with the same volume.Six rats of each group were picked up for taking blood for culture at the time points of Oh,12h,24h,and 48h after narcotization.Finally,the oprⅠ gene of each blood samples were checked with FQ- PCR.72 rats were random divided into three groups,therapeutic group,treated group and control group.Pseudomonas aeruginosa with the concentration of 1×109 CFU/ml were injected into those rats.Sensitive antibiotics,insensitive antibiotics and 0.9% NaCl were given to the therapeutic,treated and control group rats respectively.Six rats of each group were picked up for taking blood for culture at the time point of Oh,12h,24h,and 48h after narcotized.Finally,the oprⅠ gent of each blood sample were checked with FQ-PCR.Results The blood culture were positive in each period of the concentrations 1×109 CFU/ml and 1×108 CFU/ml.Results of FQ-PCR showed that the copy number decreased with time going,all of which were positive.The blood culture were positive at the time points of Oh and 12h with the concentrations of 1×107 CFU/ml and 1×106 CFU/ml,were positive with concentration of 107 CFU/ml at the time point of 24h,but negative with concentration of 107 CFU/m at the time point of 48h,and negative with the concentration of 1×106 CFU/ml at the time points of 24h and 48h.The blood culture were negative in each period of the concentration of 1×105 CFU/ml,and the results of FQ-PCR were negative.The blood culture were positive in each period of both treated and control group,but negative in each period of therhpeutic group,all the results of FQ-PCR were positive.Conclusion The coincidence rate between the method of FQ-PCR and trgditional germicuhure were 100%.Though the sensitivity of FQ-PCR was not increased,the time needed by diagnosis was shorter After treated with effective antibiotic,fhe sensitivity of FQ-PER to diagnosis Pseudomonas aeruginosa septicemia was higher than that of traditional germicuhure,and the experiment time was shorter.Detected the changes of the oprⅠ gene copies number may be helpful to estimate the sensitivity of antibiotic.

We reported a case of cryptogenic organizing pneumonia ( COP) who was admitted to the hospital in July 2008 and reviewed the Chinese literature of COP from 2003 to 2008. The most common symptoms of COP are fever, cough and exertional dyspnea. The imaging characteristics of COP are similar to those of pneumonia, therefore is often misdiagnosed as pneumonia with a high misdiagnosis rate. Lung biopsy is the main method for pathological diagnosis; polypoid growth of granulation tissue was noted within respiratory bronchioles, small airways and alveolar spaces.

Fifty-six patients diagnosed as early stage lung cancer by spiral CT scan,underwent CTguided percutaneous transthoracic biopsy with a successful rate of 100%.Forty nice cases were confirmed pathologically including adenocarcinoma in 30 cases,squamous carcinoma in 14,alveolar cell carcinoma in 2 and small cell lung cancer in 3.Three cases were diagnosed as chronic inflammatory lesion,2 cases as tuberculosis and the diagnosis was not confirmed in 2 cases.The results suggest that CT-guided percutaneous transthoracic biopsy is an effective diagnostic measure for patients with stage Ⅰ-Ⅱ lung cancer.

Objective To evaluate the early clinical efficaoy of hybrid procedure for infants less than six months old with ventricular septal defect and coarctation of aorta.Methods From January 2010 to July 2011,20 patients with ventricular septal defect and coarctation of aorta received hybrid procedure in our center.The body weight was (4.5 ± 1.6) kg ( ranged from 1.9 kg to 6.5 kg) and the age was ( 56 ± 45 ) days ( ranged from 18 days to 6 months).The pressure gradient of the coarctation of the aorta ranged from 30 mm Hg to 56 mm Hg,5 patients of them were diagnosed as hypoplasty of aortic arch.The size of the ventricular septal defect ranged from 8 mm to 16 mm.Results The mortality was zero in all the 20 cases during the surgery,and the mobidity was 20％ (4/20).The complications were pneumonia in 2 cases,infective endocarditis in 1 case and pneumothorax in 1 case.The diameter of coarctation of the aorta ranged from 1.5 mm to 3.4 mm,and the size of the balloon ranged from 4 mm to 12 mm.The pressure gradient of the coarctation of the aorta decreased to 0 to 27 mm Hg.The bypass time ranged from 40 minutes to 87 minutes,and the crossclamp time of the aorta ranged from 20 minutes to 41minutes.The atrial septal defects were repaired and the patent ductuses were ligated during the surgery without leaving the sternum open.The total operation time was (4.0 ± 0.7 ) hours ( ranging from 3.0 hours to 5.2 hours).The mean ventilation time was (2.2 ± 1.4) days and mean ICU stay time was (5 ± 3 ) days.All the patients were followed up for ( 10.0 ± 3.6) months without aneurysm in arch and obstruction in airway.The residual obstructive pressurc gradicnt in the aortic arch ranged from 12 mm Hg to 35 mm Hg and 2 patients received reintervention.One patient received re-balloon dilation and the other received surgery.The cardiac function reached NYHA Ⅰ - Ⅱ in all eases.Conclusion The early outcome of the hybrid procedure (balloon dilation of the coartation of the aorta and surgical repair of ventricular septal defect) for infants with ventricular septal defect and coarctation of aorta was satisfying,which could avoid from circulatory arrest.It is a relatively safe procedure which could be the optional method for one-stage surgical repair.

BACKGROUND:Degradable polymer materials initiate the degradation process immediately after implantation. How to regulate the degradation of these materials is rarely reported at present. OBJECTIVE:To study the effect of ultrasonic wave on control ing the degradation of polymer materials. METHODS:The sample is made ofε-caprolactone/L-lactide copolymer, and its core was coated with low density polyethylene on the surface with the fol owing four different methods. (1) The core surface was firstly covered with CaCl 2 powder, and then coated with polyethylene. (2) The core was firstly coated with polyethylene and coarsened for 3 hours. (3) The core surface was firstly covered with CaCl 2 powder, and then coated with polyethylene, and coarsened for 3 hours. (4) The core was directly coated with polyethylene. The four kinds of specimens obtained were embedded in pork for ultrasonic bombardment experiment in vitro. RESULTS AND CONCLUSION:In the specimens prepared with methods 1 and 4, the lyophobic layer could protect core materials before ultrasonic treatment, and no absorption peak was found at 631 nm. After ultrasonic treatment, the lyophobic layer was destroyed, toluidine blue dye was released, leading to change the color of immersion solution and increase the absorption peak at 631 nm. In the specimens prepared with methods 2 and 3,the lyophobic layer cannot exhibit the protection effects, the absorption peak was found at 631 nm. Under electron microscope, the appearance of the specimens in four groups was changed obviously. It is feasible to control the starting of the degradation by coating the degradable copolymer with LDPE and using ultrasonic as a trigger.

Objective To investigate the expression of Mta-1 in bladder transitional cell carcinoma (BTCC)and to analyze its correlation with the clinical staging,pathologic grading,metastasis and recur- rence,and to explore the possible molecular mechanisms.Methods Samples of 42 cases of BTCC and 12 normal bladder mueosa tissues were examined with immunohistochemical analysis for the expression of Mta- 1,ER,u-PA and PAI-1.Endothelial cells were stained by anti-CD34,and microvascular density(MVD)of carcinoma tissue was calculated.The correlation of Mta-1 expression with the invasion,metastasis,angiogene- sis and recurrence of BTCCs was analyzed;and the correlation of Mta-1 expression with ER,u-PA,PAI-1,and MVD was also analyzed.Results The positive rate of Mta-1 expression in BTCCs was 73.8%(31/42) , while it was 0.0% in normal bladder mucosa tissues(P＜0.01).The expression level of Mta-l increased with the higher clinical stages and pathologic grades of BTCCs;it was higher in recurrence group(100.0% , 15/15)than in non-recurrence group(59.3%,16/27),and high in metastasis group(100.0%,14/14) than in non-metastasis group(60.7%,17/28)(P＜0.05).The expression level of ER increased with the lower clinical stages and pathologic grades of BTCCs;the positive rate of ER expression was 0.0% in 14 ca- ses with metastasis and was 53.6% in 13 of 28 cases without metastasis(P＜0.05);and the rate was 6.7% in 1 of 15 cases with recurrence and 44.4% in 12 of 27 cases without recurrence(P＜0.05).Negative cor- relation was found between Mta-1 and ER expression(r=-0.739,P＜0.01).The positive rate of u-PA ex- pression(59.5%,25/42)was significantly higher in BTCCs than that in normal bladder mucosa tissues (16.7%,2/12)(P＜0.05).Positive correlation was found between u-PA and Mta-1 expression(r= 0.875),while negative correlation was found between u-PA and PAI-1 expression(r=-0.535).The posi- tive rate of PAI-1 expression in normal bladder mucosa tissues(50.0%,6/12)was significantly higher than that in BTCCs(19.0%,8/42)(P＜0.05).In addition,negative correlation was found between PAI-1 and Mta-1 expression(r=-0.706).And positive correlation was found between MVD in BTCCs marked by an- ti-CD34 and Mta-1 expression(r=0.683).Conclusions Mta-1 is highly expressed in BTCCs,and it correlates closely with tumor pathologic grades,clinical stages,recurrence and metastasis.Mta-1 up-regulates the expression of u-PA and down-regulates that of PAI-1,which is associated with invasion and metastasis and acts as an angiogenic mediator in BTCCs.A negative correlation is found between Mta-1 and ER in inva- sion and metastasis of BTCCs.

Objective: To evaluate the occupational health risk of key posts exposed to cement dust in four cement production enterprises, and to provide reference for cement pneumoconiosis prevention and control. Methods: Four Chinese typical cement enterprises and key posts exposed to cement dust were selected to carry out occupational health investigation and detection, and three risk assessment methods were used to assess their occupational health risk levels, including semi-quantitative comprehensive index method, semi-quantitative contact ratio method and risk rating method of International Mining and Metal Commission ( ICMM ). Meanwhile, the differences and consistencies among different assessment methods were compared. Results: Dust free silica content ranged from ( 4.70±2.01 ) % to ( 5.63±2.48 ) %,and the total and respirable dust concentrations exposed by bagged cement loaders and cement baggers exceeded Chinese permissible concentration-time weighted average（ PC-TWA ）. The results of semi-quantitative comprehensive index method showed that all the types of work were at high risk of total and respirable dust, while the results of the other two assessment methods showed that bagged cement loaders and cement baggers were at a extremely high or intolerable risk. There were no significant differences among three risk assessment methods whether in terms of total dust or respirable dust ( P>0.05 ). ICMM risk rating method and contact ratio method showed highly positive correlation in term of respirable dust ( rs=0.894, P=0.016 ), but not in term of total dust ( rs=0.733, P=0.097 ). However, the correlations of comprehensive index method with the other two methods were unable to conduct. Conclusion Bagged cement loaders and cement baggers are at high occupational health risk levels. Moreover, semi-quantitative contact ratio method and ICMM risk rating method have high positive correlation in term of respirable dust, the applicability of comprehensive index method still needs further study.

OBJECTIVETo categorize diffuse large B-cell lymphoma (DLBCL) into germinal center B cell-like (GCB) and non-germinal center B cell-like (non-GCB) subgroups by immunohistochemistry; and to investigate the underlying prognostic significance.

METHODSImmunohistochemical study for CD10, bcl-6 and MUM1 was performed on 133 cases of DLBCL. The cases were then categorized into GCB and non-GCB subgroups. The 5-year overall survival and 5-year progression-free survival rates were compared between the GCB and non-GCB groups, and among the cases with different immunohistochemical expression or with different IPI.

RESULTSAmongst the 133 case studied, CD10 was expressed in 33.1%, while bcl-6 was positive in 34.6% and MUM1 in 45.1%. CD10 expression had a favorable impact on 5-year overall survival (P=0.041) and 5-year progression-free survival (P=0.031). On the other hand, bcl-6 expression had a favor able impact on 5-year progression-free survival (P=0.044). Expression of MUM1 carried an adverse effect on 5-year overall survival (P=0.031) and 5-year progression-free survival (P=0.028). GCB immunophenotype was demonstrated in 40.6% of the cases, while 59.4% showed a non-GCB profile. GCB DLBCL had a significantly longer 5-year overall survival (P=0.004) and 5-year progression-free survival (P=0.003), as compared with the non-GCB group. When dividing the cases into two groups according to their IPI score (IPI=0 to 1 and IPI=2 to 5), it turned out that the 5-year overall and progression-free survival rates of the GCB group were significantly higher than those of the non-GCB group (P=0.019 and 0.014 respectively in cases with IPI of 0 to 1 and P=0.006 and 0.009 respectively in cases with IPI of 2 to 5). The non-GCB cases with a IPI of 2 to 5 had the poorest prognosis.

CONCLUSIONDLBCL subgrouping by immunohistochemistry and analysis of the subgrouping with IPI is feasible and useful in predicting clinical outcome.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; B-Lymphocytes ; pathology ; Child ; Child, Preschool ; DNA-Binding Proteins ; metabolism ; Disease-Free Survival ; Female ; Follow-Up Studies ; Germinal Center ; pathology ; Humans ; Immunohistochemistry ; Interferon Regulatory Factors ; metabolism ; Kaplan-Meier Estimate ; Lymphoma, Large B-Cell, Diffuse ; classification ; immunology ; pathology ; Male ; Middle Aged ; Neprilysin ; metabolism ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; Survival Rate ; Young Adult