To measure the relative affinity and type Specificity of neutralizing monoclonal antibodies against the hexon antigen of adenovirus type 7,a simple ELISA double antibody binding system has been developed.By this method,ralative affinity of nine MAbs was estimated from the antibody concentrations at approximately 50% of plateau binding and were ranked.The theoretical basis for this method was discussed. Hybridomas secreting antibodies of desired affinity can be selected at an early stage after fussion by measuring relative affinity of hybridomas supernatants.The type specificity was judged by the difference between end-point concentration against adeno- virus type 7 and type 3.And the relation between affinity and specificity of MAbs was discussed.

Objective To screen for differential expression of genes in bone marrow mesenchymal stem ceils (MSCs) stimulated by electromagnetic fields (EMFs) and to study the mechanism of differentiation of the bone mar- row MSCs induced by EMFs.Methods Mouse bone marrow MSCs were isolated and cultured in vitro.The third- passage cells were stimulated by EMFs,then the total RNA was extracted,purified to cDNA and reversely transcribed to yield a cRNA probe.The cRNA probes from stimulated and control cells were labelled with Cy5 and Cy3,respec- tively.The two samples were hybridized with a mouse oligo microarray,and the hybridization signals were scanned. The reverse transcriptase polymerase chain reaction (RT-PCR) was used to identify the interesting genes:Duspl and Rabl.Results A total of 153 differentially expressed genes were found comparing the stimulated group and the control group.There were 74 up-regulated genes and 79 down-regulated genes in the stimulated group.Semi-quantita- tive RT-PCR confirmed that the Duspl and Rabl mRNA expression levels of the stimulated group were up-regulated. Conclusion The gene expression profiles of the bone marrow MSCs were altered after EMF exposure.The genes are involved in cell proliferation and differentiation,transcription control,metabolism and response to stress.These re- sults provide deeper insight into the mechanism of differentiation of the bone marrow MSCs.

Objective To study the effects of 50 Hz magnetic fields on the in vitro proliferation of human osteosareoma cell line MG-63 with different cell densities.Methods Four different magnetic intensities(1 mT, 2 mT,3 roT,4 mT)were used to stimulate the cells,and the experiment was repeated with different cell densities. The method of MTT was employed to evaluate the level of proliferation.Results Fifty Hz magnetic fields signifi- cantly affected the level of proliferation of human osteosareoma cell line MG-63,and the 2 mT intensity exerted the greatest influence on it.The effects of the magnetic field differed with different cell densities.Conclusion The effect of 50 Hz magnetic fields on the in vitro proliferation of human osteosarcoma cell line MG-63 was not only relat- ed to the magnetic intensity,but also the cell density,

ObjectiveTo assess the safety and efficacy of retrograde ureteroscopy lithotomy (URSL)assisted antegrade percutaneous nephrolithotomy (PCNL) for complex upper ureteral calculi in semisupine-lithotomy position.MethodsFrom March 2007 to December 2010,a total of 95 patients with complex upper ureteral calculi underwent retrograde URSL assisted antegrade PCNL in semisupine-lithotomy position.Ureteral calculi size was 12 mm × 6 mm to 38 mm × 15 mm,24 cases combined with renal calculus.Firstly retrograde URSL was performed,once the stone fragments moved up to renal pelvis,a 16-22 F PCNL working channel was established under the ultrasound guidance through which lithotripsy was performed using an ureteroscope.Finally a 6-7 F double-J tube was indwelled.ResultsOperations were successfullycompleted in 93 patients.However,in it 2 patients were converted to open surgery because of significantureteral distortion due to previous open surgery.Operative time was(42.7 ± 14.9) min; estimated blood loss was(34.5 ± 26.1 ) ml.The ureteral calculi clearance rate was 100.0％,and renal calculus clearance rate inthose combined with renal calculus was 95.8％ (23/24).There were no major intraoperative and postoperative complications excepted early urinary leakage in 2 cases and fever ≥39℃ in 3 cases.ConclusionsRetrograde URSL assisted antegrade PCNL in semisupine-lithotomy position is safe and feasible for complex upperureteral calculi,especially non-opaque calculi,combined with renal calculus,easily ascending ureteral calculi and large calculi burden which has low calculi clearance rate after URSL.The outcomes are encouraging with fewer complications.It also avoids intraoperative change of patient's position.

The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.

OBJECTIVETo sum up the clinical experience in the management of benign prostatic hyperplasia (BPH) with the prostate weighing over 80 ml by transurethral resection of the prostate (TURP) combined with 2 μm continuous-wave laser vaporesection (LVR).

METHODSWe retrospectively analyzed the clinical effects of TURP combined with 2 μm LVR in the treatment of 46 cases of BPH with the prostate volume > 80 ml.

RESULTSAll the operations were successfully accomplished. The operation time and intraoperative blood loss were (112.0 ± 20.0) min (range 86-176 min) and (77.9 ± 25.9) ml (range 50-200 ml), respectively. The catheters were withdrawn at 7 days after surgery. Transient urinary incontinence occurred in 6 cases and secondary hemorrhage was found in 2 postoperatively. Six-month follow-up revealed no urethral stricture or other complications. Compared with the baseline, the international prostate symptom score (IPSS) was significantly decreased at 6 months after operation (26.3 ± 1.8 vs 11.6 ± 1.7, P <0.05), and so were the quality of life (QOL) score (5.3 ± 0.7 vs 1.3 ± 1.1, P <0.05) and post-void residual urine (PVR) ([115.5 ± 55.6] ml vs [19.9 ± 11.6] ml, P <0.05). However, the maximum urinary flow rate (Qmax) was remarkably increased from (4.1 ± 2.6) ml/s to (16.2 ± 1.7) ml/s (P <0.05).

CONCLUSIONTURP combined with 2 μm LVR is safe and effective for the treatment of BPH with the prostate volume >80 ml.

Aged ; Blood Loss, Surgical ; Humans ; Laser Therapy ; methods ; Male ; Middle Aged ; Organ Size ; Prostate ; pathology ; Prostatic Hyperplasia ; pathology ; surgery ; Quality of Life ; Retrospective Studies ; Transurethral Resection of Prostate ; methods ; Treatment Outcome ; Urethral Stricture ; Urinary Incontinence ; etiology ; Urinary Retention

OBJECTIVETo explore the effect of Compound Tongtu Granule (CTG) on intestinal permeability in elderly sepsis patients.

METHODSEighty elderly sepsis patients were randomly assigned to the experimental group and the control group by randomized double blinded method, 40 in each group. On the basis of conventional antiseptic treatment program, patients in the experimental group took CTG, while those in the control group took placebos. The dosage for CTG or placebos was 14.3 g each package, one package each time, twice daily for 14 successive days. Patients' abdominal symptoms and signs, levels of serum inflammatory factors (high-sensitivity C-reactive protein and procalcitonin), levels of plasma endotoxin, and the intestinal permeability (IP, represented by urinary lactulose/mannitol excretion rate) were compared between the two groups before and after treatment.

RESULTSAfter 14-day treatment, patients in the experimental group had improved abdominal symptoms, increased frequency of defecation, significantly decreased levels of plasma endotoxin and IP, when compared with the control group (P < 0.05).

CONCLUSIONCTG could improve the intestinal barrier function in elderly sepsis patients.

Aged ; C-Reactive Protein ; metabolism ; Calcitonin ; metabolism ; Calcitonin Gene-Related Peptide ; Defecation ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Endotoxins ; metabolism ; Humans ; Intestines ; metabolism ; Permeability ; Protein Precursors ; metabolism ; Sepsis ; drug therapy ; physiopathology

Background：Global Handwashing Day (GHD) is a handwashing promotion campaign organized by the Global Public-Private Partnership of Handwashing with Soap. In China, it has been promoted by the Chinese public health authorities, international organizations and multinational corporations through various channels including social media such as Sina Weibo, the leading Chinese microblogging site similar to Twitter. The objective of this study is to qualitatively assess Chinese social media users’ reactions to a health promotion campaign using Global Handwashing Day (GHD) 2012 as an example.Methods：We conducted a qualitative content analysis of 552 Weibo posts generated on GHD 2012 by Weibo users with 1000 or more followers with the Chinese keyword for “handwashing.” We categorized the Weibo posts into groups by keywords that frequently appeared in the data set. These groups were either exact reposts of an original post, or they conveyed similar information.Results：We observed the interconnections between traditional media and social media in handwashing promotion. Social media were found to serve as amplifiers of contents provided by traditional media. We observed the contextualization of global hygiene messages in a unique national social media market in China.Discussion：Our study showed that social media and traditional media are two interconnected arms of the GHD campaign in China. Our analysis demonstrated that public health campaigns in China can be evaluated using social media data. The themes and topics identified in this study will help public health practitioners evaluate future social media handwashing promotion campaigns.

OBJECTIVETo construct T vectors based on Xcm I recognition site and optimize the PCR fragments for its ligation.

METHODSWe firstly cloned the human histone H4 cDNA containing one Xcm I recognition site at both its 5' and 3' end into pCDNA 3.0 vector and then generated T vector with pCDNA 3.0 backbone by cutting the recombinant plasmid with Xcm I. To increase the ligation efficiency, the primers were firstly phosphorylated before DNA fragments amplification and then the PCR products were treated with Taq DNA polymerase and dATP after PCR amplification. Two DNA fragments with the length of 312 bp and 1 329 bp were ligated to it and the ligation mixture was transformed into E. coli DH5α competent cells and the positive rates of the transformants were evaluated by PCR and DNA agarose gel electrophoresis.

RESULTSOur results showed that the T vector produced by our method could ligate to the target DNA fragments with high efficiency. Besides, the phosphorylation state of the primers used for PCR amplification is also an important factor determining the cloning efficiency. What's more, as for longer DNA fragments, retreatment with Taq DNA polymerase and dATP after PCR amplification and purification could improve the ligation efficiency significantly.

CONCLUSIONOur protocol may overcome the dependence on blue/white screening to get positive clones and provide a potent way to generate T vectors and ligate them to the target PCR fragment.

Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; Genetic Vectors ; Histones ; genetics ; Humans ; Polymerase Chain Reaction ; methods

OBJECTIVETo investigate the anti hepatitis E virus (HEV) and HEV RNA in acute and convalescent sera of patients with NonA-E acute hepatitis.

METHODSThe serum samples were taken from 95 patients who were diagnosed as acute NonA-E hepatitis. Enzyme immunoassay (EIA) was used for detecting anti-HEV Immunoglobulin G (IgG, Genolable and Wantai EIA anti-HEV kits). RT-PCR amplification of HEV RNA was based on the open reading frame 2 region of HEV and the PCR products were sequenced.

RESULTSSera from 95 patients who were negative for anti-HEV in acute phase were followed up for 11-35 days to detect the anti-HEV antibody in recovery phase, 16/95 (16.84%) were positive for anti-HEV (wantai EIA anti-HEV kits). Ten (62.50%) were positive for HEV RNA in acute phase. Sequence analysis showed that 4 were HEV genotype. 6 were HEV genotype; 12/95 (12.50%) were positive for anti-HEV (Genolable EIA anti-HEV kits). Seven were positive for HEV RNA; 4 belonged to HEV genotype, 3 were HEV genotype.

CONCLUSIONIt is significant and necessary to detect anti HEV antibody and HEV RNA in patients with HEV infection during acute phase and convalesent phase.

Acute Disease ; Amino Acid Sequence ; Base Sequence ; Convalescence ; Genotype ; Hepatitis Antibodies ; blood ; Hepatitis E ; genetics ; immunology ; virology ; Hepatitis E virus ; genetics ; immunology ; Humans ; Immunoglobulin G ; blood ; RNA, Viral ; blood ; genetics ; Sequence Homology ; Viral Proteins ; genetics