Objective:To construct an adenovirus-mediated anti-sense RNA targeting K-ras exon 1 of SW1990 cell line and observe its effect on ceil proliferation and apoptosis after transferred into SW1990 cell line.Methods:K-ras exon 1 cDNA was cloned into shuttle vector pShuttle-CMV and the resultant plasmid was confirmed by enzyme digestion and PCR.Clones with inverted insertion were selected and co-transferred into E.coli BJ5183 with an adenoviral backbone plasmid pAdEasy-1 to produce recombinant plasmid by homologous recombination.Recombinants were then selected and transfected into 293 cell line to produce recombinant adenovirus.Recombinant adenovirus production was confirmed by PCR analysis and was amplified and purified;the virus titer was determined.Ad-LacZ was used to infect SW1990 cells and the infection efficiency was observed by X-gal staining.SW1990 cells was infected with the recombinant adenovirus and their proliferation and apoptosis were determined by MTT and annexin V/PI FCM assay.Results:A 282 bp target gene fragment was acquired by PCR;the titer of recombinant adenovirus was 7.6?10~8 pfu/ml before purification by CsCl_2 gradient centrifugation and 5.0?10~(10)pfu/ml after CsCl_2 gradient centrifugation.When the recombinant adenovirus was at 100 MOI,the infection efficiency of SW1990 cells nearly reached 100%.The transfection of recombinant adenovirus significantly inhibited SW1990 cell proliferation(P

OBJECTIVETo observe the clinical effects of needle-pricking therapy, a newly medical and minimally invasive technique, for functional retrograde ejaculation and to explore its mechanism. Methods Thirty-six patients with functional retrograde ejaculation were randomly divided into an observation group(19 cases) and a control group(17 cases) In the observation group,needle-pricking therapy was used at Guanyuan(CV 4) and bilateral sacral plexus nerve,lumbar 1 nerve and greater occipital nerve stimulating points,once a week. In the control group, midodrine tablets were prescribed orally,three times a day. All the treatment was given for 9 weeks. The clinical effects of the two groups were observed, and the levels of luteinizing hormone(LH), testosterone(Tes) and estra4 diol(E2) were compared between the two groups.

RESULTSThe total effective rate of the observation group was, 89. 47%(17/19), which was better than 47.06% (8/17) of the control group(P<0. 05). The LH and Tes were obviously increased and E2 was decreased compared with those before treatment in the observation group(all P< 0. 01). Tes was raised(P<0. 05) and E2 was apparently declined in the control group(P<0. 01). After treatment, the differences of serum LH and Tes were statistically significant between the two groups(both P<0. 01).

CONCLUSIONNeedle-pricking therapy has advantages for functional retrograde ejaculation probably in that stimulating lumbosacral nerves can strengthen the function of pelvic floor muscles and urethral expansion muscle and regulate sexual gland axis.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Ejaculation ; Humans ; Male ; Middle Aged ; Needles ; Sexual Dysfunction, Physiological ; physiopathology ; therapy ; Treatment Outcome

Objective:To search for a method for radical resection of pancreatic and duodenal malignancy involving the mesentery root and for the long post-operation survival of patients.Methods:From Jan.2004 to Aug.2006,a total of 26(16 male and 10 female. aged 27-70)patients with pancreatic and duodenal malignancy involving the mesentery root were treated in our department.The patients included 3 with duodenal malignancy and 23 with pancreatic malignancy.Curative resection was performed by the extended pancreaticoduodenetomy(Whipple procedure)combined with mesentery root resection(MRR)for all patients.The outcomes,safety and the post-operation survival rate were analyzed retrospectively.Results:Thirteen patients were treated with Whipple procedures combined with MRR,9 were treated with partial portal vein/superior mesenteric vein(PV/SMV)and reconstruction of the vessel,and 4 patients received pre-shunt between PV and SMV with artificial vessel graft before the extended Whipple and MRR procedures.The operation time was 2.5 to 7(4.4?1.1)hour,and blood loss was 300 to 5 000(1892?1414)ml with the blood transfusion of 0 to 5 600(2 100?1 586)ml.There was no death in our group and 7(27%)had post-operation complication.The post-operation hospital stay was 10 to 30 days.The pathologic examination showed negative surgical margins for all specimens.The tumor size was 4 to 10 (6.17?2.03)cm.After a follow-up of 9 to 38 months,the pain was relieved in all patients.One of the 3 patients with duodenal adenocarcinoma had liver metastasis at 10 months after operation,and the other 2 survived 10 months and 27 months without evidence of tumor reccurence.The patient with pancreatic micro-adenocarcinoma died of local reccurence 9 months after operation.The patient with neuroendocrine carcinoma died of organ failure 24 months after operation.The patient with lymphoma have survived for 24 months after operation.The 1-year and 2-year accumulated survival rates in the 20 cases with pancreatic ductal cancer were 86.6% and 45.6%. respectively.Conclusion:The extended Whipple procedure with MRR is safe and effective.It can obtain R0 resection in patients with malignant tumors(over 5 cm in diameter)in the head,neck and uncinate process of the pancreas and duodenal.

OBJECTIVETo study the cultural characteristics of Armillaria mellea (A. mellea ) on solid media.

METHODSA. mellea was cultured on semi-solid agar medium in dark conditions. Effects of different media, carbon sources, nitrogen sources, and temperature on growth and morphology of A. mellea were observed. The contents of polysaccharide, mannitol, glucose, and reducing sugars in A. mellea during different stages of development were determined.

RESULTSThe biomass and morphology of A. mellea were different in various media. Sugars were more effective carbon sources than the relevant sugar alcohols. Little molecular carbon sources such as alcohol and glycerol could be utilized by A. mellea, but starch only could be utilized slowly. Either organic or inorganic nitrogen sources could be uptaken and utilized effectively by A. mellea. No evidence was found that VitB1 affects the growth of A. mellea. The growth cycle on wort medium at 30 degrees C was shorter than that at 25 degrees C for 7 days. In logarithmic growth phase and stable phase, the polysaccharide contents of A. mellea were 9.24% and 4.70% respectively, while the mannitol contents were 10.08% and 10.58% respectively; glucose and reducing sugar contents remained low level in the whole growth stage.

CONCLUSIONSCarbon sources have a more remarked effect on the growth of A. mellea than the nitrogen sources do. Optimal temperature for the growth of A. mellea ranges 20-30 degrees C. Mannitol accumulates more than other little molecular carbohydrates in A. mellea.

Agaricales ; chemistry ; growth & development ; Culture Media ; Mannitol ; analysis

Adult ; Air Pollutants ; adverse effects ; Humans ; Male ; Military Medicine ; Monitoring, Physiologic ; Noise, Occupational ; adverse effects

OBJECTIVEThe Chinese Children's Leukemia Group (CCLG)-acute lymphoblastic leukemia (ALL) 08 protocol for childhood ALL was established in 2008. This study aims to evaluate the drug-related toxicities of CCLG-ALL 08 protocol in the treatment of childhood ALL.

METHODSA total of 114 children with newly diagnosed ALL were treated with the CCLG-ALL 08 protocol. The protocol was divided into five phases: remission induction (VDLD), early reinforcement (CAM), consolidation therapy, delayed reinforcement (DIa & DIb) and maintenance treatment. Drug-related toxicities in each phase were evaluated according to the Common Terminology Criteria for Adverse Events version 4.0.

RESULTSToxicities were more frequent in phase VDLD than other treatment phases, including hepatotoxicity (87.7%), dental ulcer (20.2%), hyperglycemia (20.2%), prolonged activated partial thromboplastin time (21.1%) and decreased fibrinogen (34.2%), with the incidence rates of severe adverse events at 7%, 0, 1.3%, 0.8% and 2.7% respectively. The incidence of allergic reaction to L-ASP was significantly higher in phase DIa than in phase VDLD (28.0% vs 7.9%; P<0.01), and there were no longer any allergic reactions in 15 patients who received continuing treatment with pegaspargase instead. There was no severe arrhythmia, myocardial ischemia, decreased left ventricular function, osteonecrosis, myopathy, organ failure or treatment-related mortality.

CONCLUSIONSThe drug-related toxicities of CCLG-ALL 08 protocol are common in phase VDLD, but they are mild and reversible. There is no treatment-related mortality. The CCLG-ALL 08 protocol for childhood ALL is safe.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; Asparaginase ; adverse effects ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Remission Induction

OBJECTIVEBy means of the detection of the numbers of CD34(+) cells and eosinophils (EOS), and the level of IL-5 in peripheral blood from normal controls and patients with allergic rhinitis pre- or post-treatment, the role of EOS-stem cells paths for treatment effect in allergic rhinitis (AR) was studied so as to find the convenient and quick indicators which could be used to evaluate the clinical therapeutic effect and adjust the methods of the hormone therapy.

METHODSThere were 2 groups. (1) experimental group: 44 patients, including 24 males and 20 females with a age range of 7 to 68 years old. The patients received the treatment of fluticasone spurt for four weeks. (2) CONTROL GROUP: 30 cases, from normal health examination. The EOS numbers, CD4(+) cell numbers and the IL-5 level were examined in control group as well as before and after therapy in experimental group.

RESULTSThe IL-5 level and CD numbers before therapy in experimental group were (88.25 +/- 33.47) ng/L, (9.24 +/- 2.15)/10(5), significantly higher than that after therapy and experimental group which were (44.34 +/- 16.32) ng/L, (6.31 +/- 1.83)/10(5) and (31.24 +/- 8.43) ng/L, (3.47 +/- 1.32)/10(5) respectively. The IL-5 level was positively correlated with the CD34(+) number pre-and post treatment in experimental group (r = 0.64, P <0.01; (r = 0.61, P <0.01). The EOS number was positively correlated with the level of IL-5 (r = 0.64, P <0.01).

CONCLUSIONSIL-5 and CD34(+) cells can be regarded as indicators to evaluate the therapeutic effect. The EOS, CD34(+) cells and the level of IL-5 in the peripheral blood are correlated with the pathogenesis of AR, suggesting that there is a related path between the local nasal tissue of AR patients and the marrow.

Adolescent ; Adult ; Aged ; Androstadienes ; therapeutic use ; Anti-Allergic Agents ; therapeutic use ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; Case-Control Studies ; Child ; Eosinophils ; immunology ; Female ; Fluticasone ; Humans ; Interleukin-5 ; blood ; Male ; Middle Aged ; Rhinitis, Allergic, Perennial ; blood ; drug therapy ; Stem Cells ; Young Adult

Objective To explore a sensitive method and index to evaluate renal functional reserve (RFR) in patients with early diabetic nephropathy (DN) using protein loading-scintirenography.Methods Fifty subjects were studied and divided into 3 groups.Group one (G1) consisted of 14 healthy volunteers;Group two (G2) consisted of 15 patients with type 2 diabetes mellitus (DM) and normoalbuminuria; Group three (G3) consisted of 21 patients with type 2 DM and microalbuminuria.All subjects underwent baseline and protein loading-99 Tcm-DTPA scintirenography within one week.RFR was calculated as the difference between stimulated and baseline glomerular filtration rate (GFR), time of peak ( Tb ), time of half excretion ( C1/2 ), residual rate at 20 min ( C20/b ) .Variance analysis and t-test were used to analyze the group differences.Results ( 1 ) The RFR in terms of GFR had statistical difference between any two groups (t=14.884, 32.180, 16.042, all P＜0.01).After protein-loading, the GFR of G1, G2 and G3 increased 20.1, 10.9 and 2.2 ml·min-1·1.73 m-2 respectively.Therefore, the RFR decreased before microalbuminuria in type 2 DM patients.(2)There was statistical difference between the RFR of G1 and G2 in terms of C1/2 (t = 5.505, P＜0.05 ), and between G1 and G3 ( t = 8.914, P＜0.01 ).(3) There was statistical difference of the RFR in terms of TP between G1 and G3 (t = 5.690, P ＜ 0.01 ).(4) There was statistical difference of the RFR in terms of C20/b between G1 and G3 (t= 4.376, P＜0.05 ).Conclusions 99Tcm-DTPA protein loading-scintirenography is an effective method for measuring RFR to evaluate early DN in type 2 DM patients.

This paper is to explore the reasonable applications of automatic exposure control in computed radiography, and to improve the quality of CR images. It is very important to select a suitable KV value in automatic exposure control in computed radiography. At the same time, a suitable ionization chamber, correct density compensation and necesary post-processing should be selected.
Adolescent ; Adult ; Aged ; Automation ; Child ; Child, Preschool ; Female ; Head ; diagnostic imaging ; Humans ; Image Processing, Computer-Assisted ; Infant ; Lumbar Vertebrae ; diagnostic imaging ; Male ; Middle Aged ; Quality Control ; Radiation Dosage ; Radiographic Image Enhancement ; methods ; Radiography, Thoracic ; Tomography, X-Ray Computed ; methods ; standards

To construct Fv antibodies against H5N1 Avian influenza virus hemagglutinin,extracted mRNA from B lymphoblastoid cell lines secreting anti-HA antibodies was used and the VH and VL genes were amplified by RT-PCR and linked together by splicing overlap extension (SOE) with (Gly4 Ser)3 linker. The recombinant plasmid was then transformed to E. coli BL21(DE3) and sequence analysis indicated the total length of scFv was 714 bp and the expression of Fv was validated by PAGE and Western blot.
Animals ; Antibodies ; genetics ; metabolism ; pharmacology ; Birds ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression Regulation ; Hemagglutinins ; immunology ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; metabolism ; Influenza A Virus, H5N1 Subtype ; drug effects ; immunology ; Influenza in Birds ; virology ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; genetics ; metabolism ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Proteins ; immunology