Object To establish a calli culture system for the rapid propagation of Speiraea japonica L. f.. Methods Callus and shoot induction were carried out on MS, 6, 7 V or B 5 media with different parts of the plant such as stem tip, tender leaves and petiole as explants. Results A calli culture system was established for the rapid propagation of S. japonica. Conclusion MS cultural medium was found to be most suitable for calli induction. MS with 2.0 mg/L 2,4 D+0.3 mg/L KT can induce calli when the explants were used for the induction, with stem tips being the most satisfactory explant. Clusters of seedlings can be induced on MS+2 mg/L BA+0.1 mg/L NAA and when these seedlings were transferred to 1/2 MS +0.25 mg/L BA+0.5 mg/L NAA medium, root were developed to give young seedlings.

BACKGROUND:There is few mesenchymal stem cells(MSCs) in the bone marrow,about one BMSC in 1?104-1?105 monocytes.Following in vitro isolation,purification and amplification,it is satisfactory for in vivo requirement.OBJECTIVE:To observe biological characteristics and potentiality of multi-directional differentiation of BMSCs in vitro.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Institute of Urinary Surgery,Union Hospital Affiliated to Fujian Medical University from October 2007 to December 2008.MATERIALS:A total of 30 clean male Sprague Dawley rats of inbreeding line were supplied by Silaike,Shanghai,China.METHODS:BMSCs from rats were isolated and cultured in vitro by using the whole bone marrow adherence method.When 80%-90% confluency,BMSCs were digested by trypsin.At the third passage,BMSCs were induced to differentiate into osteoblasts and adipocytes.MAIN OUTCOME MEASURES:Cell morphology was observed under the inverted phase contrast microscope.BMSCs surface markers were detected using flow cytometry.Osteogenic ability was examined by alizarin red staining.Adipogenic ability was measured by Oil red O staining.RESULTS:The primary cells and the passage cells were mostly fusiform in shape,to be similar to fibroblasts.Cell still kept a high potential of growth and amplification following over 10 subcultures.At the third passage,BMSCs were positive for CD44,CD90,CD106,but negative for CD34,CD45,CD11b.Following 21 days of osteogenic induction,cell alizarin red staining showed that alizarin red was positive in osteoblasts.Following 2 weeks of adipogenic induction,oil red O staining showed that red lipid droplet existed in adipocytes.CONCLUSION:Whole bone marrow adherence method can isolate,purify and amplify BMSCs in vitro.The obtained cells have general biological characteristics of MSCs,and also have potentiality of multi-directional differentiation.

Objective To investigate the expressions of fatty acid-binding protein 5 (FABP5)and dihydroli-poamide dehydrogenase(DLD)in skin lesions of symmetrical acrokeratoderma(SAK), and to explore their significance. Methods Biopsy specimens were obtained from skin lesions on the wrists and perilesional skin of 9 patients with SAK, and from normal skin in the wrists of 9 healthy volunteers (control group). Reverse transcription PCR (RT-PCR)and immunohistochemical staining were performed to measure the expressions of FABP5 and DLD in these specimens. Results RT-PCR showed no significant differences in the mRNA expressions of FABP5 or DLD between lesional, perilesional and normal control skin specimens(both P > 0.05). Immunohistochemically, there was a significant increase in the extent and intensity of staining for FABP5 in SAK lesions. Concretely speaking, FABP5 was strongly expressed in the stratum corneum, granular and spinous layers in SAK lesions, but weakly expressed in the stratum corneum, granular and spinous layers in perilesional skin, and only in spinous and basal layers in normal control skin. The expression of DLD decreased in SAK lesions, and was observed only in the stratum corneum and spinous layer in a few cases of SAK. However, the full-thickness epidermis stained positive for DLD in perilesional skin, with the nuclei and cytoplasm both stained deep brown. Conclusion The overexpression of FABP5 in SAK lesions may participate in dysdifferentiation of keratinocytes, while the down-regulation of DLD expression suggests an imbalance in energy metabolism.

Objective To study the feasibility of reducing radiation dose in aorto-iliac and lower extremity arteries CT angiography (CTA) with low tube voltage (100 kV) and automatic tube current modulation(ATCM).Methods Totally 61 patients requiting aorto-iliac and lower extremity arteries CTA for clinical reasons were prospectively enrolled in study.The patients were randomly assigned to 3 groups:Group A(120 kV),Group B (100 kV) and Group C (100 kV with automatic tube current modulation).Both quantitative and qualitative analysis were included in this research.Group C was divided into obese group(BMI≥ 24.9 kg/m2) and normal group(BMI ＜ 24.9 kg/m2).The radiation doses were analyzed respectively among two groups.Results The subjective evaluations of image quality for axial,MIP and VR were good.There were no significant differences among group A,B and C in SNR and CNR (P ＞0.05).The effective dose of group A,B and C was 8.20 mSv,5.36 mSv,and was 7.48 mSv,respectively Group B was 34.6％ less than group A.Group C was 39.5％ more than group B,and there was no significant differences between group A and C (P ＞ 0.05).The effective dose of group C1 was 7.11mSv,group C2 was 9.69 mSv,the E with group C1 were significantly less than group C2 (t =-3.163,P ＜0.05),the effective dose of group C1 was 13.3％ less than group A (Z =-2.822,P ＜ 0.05),but the group C2 was more than group A (P ＞ 0.05) and group B (Z =-3.426,P ＜ 0.05).Conclusions Lowkilovoltage (100 kV) CT scanning protocol is feasible in multi-detector CT angiography for aortoiliac and lower extremity arteries.Automatic low tube voltage (100 kV) with automatic tube current modulation (ATCM) scanning protocol can be used for someone with BMI less than 24.9 kg/m2.

Objective To investigate the effects of CD4+T cell depletion in BALB/c mice on the immunogenicity and virulence of replication-competent recombinant vaccinia virus. Methods Twelve BALB/c mice were inoculated with recombinant Tiantan vaccinia ( rTV, n=8 ) or vaccinia virus Tiantan strain ( VTT, n=4) by tail scarification after the depletion of CD4+T cells with anti-CD4 monoclonal anti-body( McAb) injected intraperitoneally for three days before virus inoculation.A control group without anti-body treatment was set up accordingly.Several parameters including the body weight, pocks on tail, viral shedding and the percentage of CD4+T cells were monitored.In the fourth week after virus infection, ovaries were collected from mice and viral loads in the tissue were titrated by plaque forming assay on chick embryo fibroblast ( CEF) cells.The specific cellular immune responses against vaccinia virus and HIV induced by inoculation of VTT and rTV respectively were detected by intracellular cytokine staining ( ICS) assay.ELISA was used to detect antibodies against vaccinia virus and HIV-1 gp120.Results All mice with or without CD4 McAb treatment showed typical poxvirus pocks on tails after inoculation of vaccinia viruses, but none of them developed secondary or satellite lesions.It took a longer time for CD4+T cell-depleted mice to heal from lesions, to regain body weights and to release viruses than the mice in control group.No vaccinia virus was detected in the ovaries of CD4+T cell-depleted mice or mice in control group.The mean absorbance( A) values for the detection of HIV-specific and vaccinia virus-specific antibodies in CD4+T cell-depleted mice with ELISA were respectively 0.119 and 0.168, which were significantly lower than those in mice of control group.The titers of neutralizing antibodies against vaccinia virus in McAb/rTV treated mice (1 ∶321) were lower than those in rTV treated mice (1 ∶1286) (P<0.05).The percentages of CD4+T cells secreting IFN-γ(0.654%) in McAb/rTV treated mice were significantly lower than those in rTV treated mice in the fourth week after immunization (P <0.0004).No significant differences with the vaccinia virus-specificCD8+ T cell responses were observed among mice with or without CD4+T cells depletion.Conclusion Thereplication and dissemination of replication-competent recombinant vaccinia virus could be effectively controlledin the mice with CD4+ T cell-depletion.The depletion of CD4+ T cells significantly reduced the humoraland CD4+ T cell responses, but had no effect on CD8+ T cell responses.

Objective To investigate the diagnostic value of the T-SPOT.TB in connective tissue disease(CTD) combined with tuberculosis.Methods This is a case-control study.Forty-four patients with CTD combined with tuberculosis were enrolled from Xiangya Hospital of Central South University from September 2011 to July 2012.Another forty-four CTD patients without tuberculosis were evaluated as a control group.The diagnostic value of T-SPOT.TB and risk factors of the false negative results by T-SPOT.TB were analyzed.Results The sensitivity of T-SPOT.TB (70.5％,31/44) was significantly higher than that of TST(27.3％,12/44) for CTD combined with tuberculosis patients (x2 =16.42,P ＜ 0.001).The specificity of T-SPOT.TB and TST were 93.2％ (41/44) and 88.6％ (39/44),respectively.There was no significant difference between the specificity (x2 =0.14,P =0.711).The positive predictive value of T-SPOT.TB was 91.2％ (31/34).The negative predictive value was 75.9％ (41/54).Youden's index was 0.64,and the positive likelihood ratio was 10.3.All the index were higher than that of TST (0.16 and 2.4).While the negative likelihood ratio which was 0.32 was lower than that of TST (0.82).When spot forming cell frequencies of T-SPOT.TB of PBMC was set to 38SFCs/106 PBMC,it had the best cut-off value.Age,use of glucocorticoids or immunosuppressant therapy,lymphocytopenia and hypoalbuminemia were not associated with false negative T-SPOT.TB assay.Conclusion The T-SPOT.TB assay is much more useful than TST for diagnosing CTD combined with tuberculosis.

To study the chemical constituents of Lysimachia patungensis Hand.-Mazz., silica gel column chromatography, reverse phase ODS column chromatography, MCI and Sephadex LH-20, were used to separate the 95% EtOH extract of the whole plant of Lysimachia patungensis Hand.-Mazz.. The structures of the isolated compounds have been established on the basis of chemical and NMR spectroscopic evidence as well as ESI-MS in some cases. Twelve phenolic compounds were obtained and identified as quercetin-3, 3'-di- O-alpha-L-rhamnoside (1), myricetrin (2), quercitrin (3), rutin (4), 2-hydroxynaringenin-4'-O-glucopyranoside (5), naringenin 7-O-glucopyranoside (6), liquiritin apioside (7), licochalcone B (8), tetrahydroxymethoxy chalcone (9), methyl-p-coumarate (10), 2, 4, 6-trihydroxy acetophenone-2-O-glucopyranoside (11) and vaccihein A (12). Among them, compound 1 is a new compound, and compounds 5, 11 and 12 are isolated from the genus Lysimachia L. for the first time, and the others are isolated from the plant for the first time.

Objective To evaluate the therapeutic effects and complications of modified facial mask for non-invasive ventilation (NIV) in elderly patients with respiratory failure.Methods A total of 132 elderly patients(107 males and 25 female,aged 78.5±8.6 years) treated with NIV from February 2008 to May 2011 were randomized into two groups:modified facial mask(group A,n=68,56 males and 12 females,aged 78.8±22.2 years) and control facial mask(group B,n=64,64 males and 13 females,aged 76.6±20.4 years).Duration of NIV,time in RICU(respiratory intensive care unit),length of hospital stay,risk for hospital acquired pneumonia (HAP),risk for invasive ventilation,cure rates,in-hospital mortality,NIV failure rate and cost were compared between the two groups.The complications of NIV,such as oropharyngeal dryness,skin damage of face and nose,abdominal bloating,gas leakage from mask were also compared between the two groups.Results Compared with group B,duration of NIV(12.2±2.3 d vs.18.4±3.6d),time in RICU(7.3±3.2d vs.14.6t5.4d),length of in hospital stay(16.6±4.2d vs.28.2±6.2)d,and cost(2.23±0.12 ten thousand yuan vs.4.23± 0.24 ten thousand yua) in group A were significantly decreased(t=9.72,14.91,13.08,10.81 respectively,all P＜0.05).The risk for invasive ventilation [2.9％ (2 cases) vs.43.8％(28 cases)],NIV failure rate [5.9％ (4 cases) vs.12.5％ (28 cases)] were also decreased in group A compared with group B(x2 =31.26,25.74,both P＜0.05).Compared with group B,The complications of NIV such as skin damage of face and nose[4.4％ (3 cases) vs.37.5％ (24 cases)],abdominal bloating [2.9％ (2 cases) vs.28.1％ (18 cases)],gas leakage from mask [8.8 ％ (6 cases)vs.50％(32 cases)] in group A were significantly decreased(x2 =31.26,25.74,all P＜0.05).Conclusions Modified facial mask for NIV is effective in the treatment of elderly patients with respiratory failure.The complications and in-hospital mortality are reduced with the application of modified facial mask for NIV and it is highly tolerated by patients.Modified facial mask for NIV is the first choice in the treatment in elderly patients with respiratory failure.

ObjectiveTo assess the cause of tadalafil failure and the feasibility of successfully rechallenging nonresponding patients.Methods A total of 80 consecutive erectile dysfunction ( ED ) patients who claimed poor response to tadalafil were enrolled into the study.A self-administered tadalafil-use questionnaire composed of eight questions was applied to assess how they had used tadalafil.Subjects were given thorough instruction based on individual answers and four doses of tadalafil 20mg.After a 2-week follow-up,end point efficacy of rechallenge was evaluated using the sexual encounter profile (SEP),which was recommended by international advisory panel in 2004.ResultsA total of 45 subjects had one or more areas of major suboptimal use of tadalafil:21.2％ did not know that sexual stimulation was necessary for tadalafil to work,87.5％ attempted to use tadalafil less than four times,57.5％ took a maximal dose less than 20 mg,and 84％ felt nervous or anxious.Of the 65 patients undergoing tadalafil rechallenge,30 patients answered “yes” to SEP2 and SEP3.The response rate to rechallenge was 46.2％ ( 30/65 ).ConclusionsInappropriate use of tadalafil was major cause of tadalafil non-pesponse.The efficacy of tadalafil could be improved to a better extent by education of patients.

Building up a harmonious type of physician-patient relationship in hospitals is an important contents in the construction of harmonious society.Major factors that jeopardize the physician-patient relationship include insufficiency of mutual communication,lack of human sympathy in medical treatment service,neglect of patients' right of informed consent.This article also puts forward some measures to solve these contradictions,such as reinforcing ethical training,improving physician-patient relationship,respecting patients' right of informed consent,and setting a psychological department for concerned issues,etc.