OBJECTIVE:To study improvement effect of modified mahuang chanyi decoction on allergic rhinitis(AR)model guinea pigs and its mechanism. METHODS:32 guinea pigs were randomly divided into normal group (0.9% Sodium chloride in-jection),model group (0.9% Sodium chloride injection),experimental [Modified mahuang chanyi decoction,5.18 g (medicinal materials)/kg] group and control [Fluticasone propionate nasal spray,50 μg/(guinea pig·d)] group. Except for normal group,other groups received ovalbumin 30 mg and calmogastrin 30 mg intraperitoneally to induce AR model. Except for control group,other groups were given relevant medicine intragastrically,twice a day for consecutive 15 d. Nasal symptoms of guinea pigs were ob-served and recorded,and then analyzed and scored. The protein expression of nasal mucosa SHP-1 was detected by immunohisto-chemistry,the serum contents of IL-4 and IFN-γ were determined by enzyme-linked immunosorbent assay. RESULTS:Compared with normal group,model group suffered from AR symptom,and nasal symptoms score was increased;and the protein expression of nasal mucosa SHP-1 and the serum content of IL-4 increased,while the content of IFN-γ decreased(P<0.05). Compared with model group,AR symptom of experimental group and control group were improved;nasal symptoms score,the protein expression of nasal mucosa SHP-1 and the serum content of IL-4 decreased while the content of IFN-γ increased(P<0.05). CONCLUSIONS:Modified mahuang chanyi decoction shows certain improvement effect on AR,and its mechanism is associated the decrease of IL-4 content and the increase of IFN-γcontent in serum.

Objective To explore the effect of PDCA circulation on the training of clinical nursing teachers.Methods PDCA circulation was applied to the training of clinical nursing teachers byPP(planning),D(doing),C(Checking),A(acting). The results of theoretical exams,teaching skills of teachers and the student’s satisfaction with clinical teaching were compared.Result The application of PDCA circulation in clinical nursing teachers training significantly improved the theoretical exam results and teaching skills of the teachers, increased student’s satisfaction with the ways and the skills of teaching,compared to the results before its use(P<0.05).Conclusions The application of PDCA circulation in training clinical nursing teachers can effectively improve the ability of clinical teachers and increase the student’s satisfaction with teachers,thus improve the quality of nursing teaching.

AIM: To activate hepatic stellate cells (HSC) in vitro in precision-cut liver slices (PCLS) stimulated by acetaldehyde for studying and screening anti-fibrotic drugs. METHODS: PCLS were prepared by the vibratome, and incubated with 700 ?m?L~-1 acetaldehyde for 0, 2, 4 and 6 h. The medium and homogenate were retained to determine glutathione S-transferase (GST) activity, lactate dehydrogenase (LDH) leakage and hydroxyproline (Hyp) content. PCLS were prepared for paraffin sections. The expression of ?-smooth muscle actin (?-SMA) was evaluated by immunohistochemistry, and the result was analyzed with image analysis software. RESULTS: The leakages of GST and LDH were increased significantly compared with those in 0 h group (P

Objective To investigate the genotype distribution of extended-spectrum?-lactamases(ESBLs) and AmpC?-lacta- mases produced by Escherichia coli and Klebsiella pneumoniae in 10 teaching hospitals of China.Methods 90 clinical strains of E.coli and 61 strains of K.pneumoniae isolated in 2003 and confirmed to produce ESBLs were collected from 10 teaching hos- pitals in China.Analytical isoelectric focusing was used to measure the pI of the?-lactamases.Conjugation experiment was used to study the transfer of cefoxitin resistance.Plasmid-mediated AmpC enzyme genes were amplified and sequenced by multiplex polymerase chain reaction (MPCR).Results The prevalence of ESBL-producing E.coli and K.pneumoniae was about 50% in Wuhan,Nanjing and Jinan.The prevalence of ESBL-producing E.coli was lower than K.pneumoniae in Beijing.However,in other hospitals the prevalence of ESBL-producing E.coli was a little higher than K.pneumoniae.About 24.4% of ESBL-pro- ducing E.coli isolates and 19.4% of ESBL-producing K.pneumoniae isolates were resistant to cefoxitin.Cefoxitin-resistant i solate was identified in all hospitals except Shenyang.Major genotype of ESBL-producing isolates was CTX-M.The CTX-M-9 group was the most common group,followed by CTX-M-1.More K.pneumoniae isolates produced both ESBLs and AmpC en- zyme than E.coli.The genotype was CTX-M/DHA-1.The PCR results of 3 transconjugants producing both ESBLs and AmpC enzyme were the same as their donor isolates.Conclusions The genotype of ESBL-producing isolates is mainly CTX-M-9 group in these teaching hospitals.More K.pneumoniae isolates produced both ESBLs and AmpC enzyme than E.coli.Most of these isolates are due to geno type CTX-M/DHA-1,which can spread through plasmid.

Objective To evaluate the feasibility and safety of taking arbitrary position in the early period after laparoscopic cholecystectomy.Methods 186 patients who underwent laparoscopic cholecystectomy from Jun.2010 to Oct.2010 were randomly divided into two groups,observation group (n =94) and control group ( n =92) ; the former group was instructed to freely mobilize 2 hours after the operation,while the latter group was administered with traditional nursing.Abdominal distension,pain,urinary retention of both groups after laparoscopic cholecystectomy were compared.Results The observation group happen abdominal distension 4 cases,urinary retertion in 3,lumbar muscle ache 15 cases and control group happen abdominal distension 13 cases,urinary retention in 12 cases,lumbar mascle ache 60 cases,and muscle soreness after laparoscopic cholecystectomy were obviously lower in the observation group,and the differences between both groups were statistically significant ( x2 =5.460,6.087,46.885 ; P ＜ 0.05 ).Conclusions Taking arbitrary position in the early period 2 hours after laparoscopic cholecystectomy is safe and feasible,it can also reduce the occurrence of abdominal distension,urinary retention and muscle soreness,mitigate pain,dizzy,palpitation and lack of energy,and facilitate patients' early recovery.

A simple and selective ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) assay was developed for the determination of the human plasma protein binding of four bioactive flavonoids (such as orientin and vitexin) in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C18 column with a mobile phase composed of 0.1%formic acid in acetonitrile and 0.1%aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITY?TQD system was operated under the multiple reaction monitoring (MRM) mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations (r40.99) of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.

Objective To investigate the effect of ischemic preconditioning (IPC) on the expression of a disintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS-1), and to study whether the application of small interfering (si)RNA specifically targeting ADAMTS-1 would help to recover IPC protection in the aged heart. Methods The 32 young (4 months) and 32 aged(24 months) male Sprague-Dawley (SD) rats were assigned randomly to IPC group (n=20) and sham operated group (n= 12) respectively. Myocardial samples from the ischemic-reperfused region were harvested for detecting the ADAMTS-1 expression. In addition, the 110 aged SD rats were assignedrandomly to ADAMTS-1 siRNA group and control group (n=55, each). The effects of ADAMTS-1siRNA transfcction on the expression of ADAMTS-1 protein, myocardial infarction survival rate,heart function and myocardial infarction size after IPC were observed.Results Twenty-four hours after IPC, the ADAMTS-1 protein expression increased significantly in iscbemic-reperfused region both in young and aged rats (P＜0. 05), and the protein expression was higher in aged rats than in young rats (P＜0.05). In young-IPC group, the absorbency showed ADAMTS-1 protein expression at 0 hrs and 24 hrs after IPC were 0. 05±0.01 and 0.12±0.03 by immunohistochemical staining, and were 0.68±0. 16 and 1. 17±0.21 by Western blots respectively. In aged-IPC group, the absorbency showed ADAMTS-1 protein expression at 0 hrs and 24 hrs after IPC were 0.07±0. 03 and 0.21 ±0.04 by immunohistochemical staining, and were 0. 76±0. 21 and 1. 48±0. 17 by Western blots. In the aged rats, ADAMTS-1 siRNA transfection inhibited ADAMTS-1 protein expression (0. 66±0. 19and 0.78±0.21, by Western blots at 0 hrs and 24 hrs after IPC, P＞0.05), but didn't improve myocardial infarction survival rates [ADAMTS-1 siRNA group and sham operated group: 14.3% (5/35) vs. 17.1 %(6/35), P＞0.05], left ventricular fractional shortening [(14.0±3.2)% vs. (13.0±2.9)%, P＞0.05] and myocardial infarction size[(39.0±4.1)% vs. (38.0±5.3)%, P＞0.05].Conclusions ADAMTS-1 expression induced by IPC increases significantly in aged versus in young rats. ADAMTS-1 knockdown by siRNA inhibits ADAMTS-1 protein expression but cannot recover the age-associated loss of IPC protection.

OBJECTIVETo observe the change in the amount of sialic acids on hepatocellular carcinoma (HCC) cell membrane.

METHODSSurgical specimens of HCC and liver cirrhosis tissues were obtained from 28 patients to prepare carcinoma cell and hepatocyte suspensions by collagenase digestion. For assay of α2, 3 and α2, 6-sialic acids, the cells were suspended in the staining buffer containing either fluorescein isothiocyanate-Maackia amurensis lectin (FITC-MAL) or fluorescein isothiocyanate-Sambucus nigra bark lectin (FITC-SNA) and incubated for 1 h, respectively. Flow cytometric analysis was carried out to measure the mean fluorescence intensity (MFI) on the cell surface.

RESULTSIn both FITC-MAL- and FITC-SNA-incubated HCC cells, the MFI on the cell surface was greater than that of the hepatocytes.

CONCLUSIONBoth of α2, 3 and α2, 6- sialic acids increases significantly on the hepatocyte membrane after the carcinomatous change.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Membrane ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Sialic Acids ; metabolism

OBJECTIVETo explore practical protocols for cloning bone marrow-derived hepatic stem cells in vitro.

METHODSThe cell fraction rich in CD117(+) cells and CD184(+) cells was separated from fresh bone marrow by density gradient centrifugation and cultured for 0, 7 and 14 days in high-glucose DMEM supplemented with or without 10% autologous serum or in serum-free high-glucose DMEM. All the media were supplemented with different concentrations of hepatocyte growth promoting factors (HGPF), thrombopoietin (TPO) and interleukin-3 (IL-3). The quantitative changes of CD117(+) cells and CD184(+) cells were measured by flow cytometry.

RESULTSThe optimal effect for cell cloning was achieved with high-glucose DMEM with 10% autologous serum group supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3. At day 7 of cell culture in this media, the quantity of CD117(+) cells and CD184(+) cells increased by 6.55 and 6.20 folds, and by 11.62 and 20.57 folds at day 14, respectively.

CONCLUSIONIt is practical for cloning bone marrow-derived hepatic stem cells in high-glucose DMEM with 10% autologous serum supplemented with 40 microg/ml HGPF, 50 ng/ml TPO, and 10 ng/ml IL-3.

Bone Marrow Cells ; cytology ; Cell Culture Techniques ; Clone Cells ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; physiology ; Humans ; Liver ; cytology ; Proto-Oncogene Proteins c-kit ; metabolism ; Stem Cells ; cytology ; Thrombopoietin ; pharmacology

Hematopoiesis is coordinated by a complex regulatory network of transcription factors that involves proliferation, differentiation and maturation of a very small population of pluripotent hematopoietic stem cells with self-renewing and differentiating into various specialized and distinct blood cell types. Malfunction of transcription factors may lead to diseases such as acute myeloid leukemia (AML). The purpose of this study was to investigate the expression pattern of transcription factor mRNA in acute myeloid leukemia (AML) cells during in vitro differentiation. The 2 human leukemic cell lines HL-60 and NB4 had been used as model cell lines. Differentiation of HL-60 and NB4 cells was induced by all-trans retinoic acid (ATRA) for 4 days. Morphological changes were observed by May-Grunwald Giemsa stainings, the CD11b expression level was detected by flow cytometry. Transcription factor mRNA profiles (PU.1, C/EBPα, ε, γ, GATA-1, GATA-2) were determined by real time RT-PCR during in vitro HL-60 and NB4 differentiation; The expression level of transcription factor mRNA was relatively quantitatively analyzed by using 2(-ΔΔCT) and compared with control group. The results showed that the expression levels of PU.1 and C/EBP ε mRNA in NB4 differentiation group were 5.75 and 6.16, respectively, which were significantly higher than those in untreated group; while the expression level of C/EBPα, γ, GATA-1, GATA-2 mRNA in NB4 differentiation group were 62%, 31%, 63% and 8.7% respectively, which were significantly lower than those in untreated group; In HL-60 differentiation group, the expression levels of PU.1, C/EBPα, ε were 1.97, 1.95 and 2.35 respectively, which were significantly higher than those in untreated group; while the expression levels of C/EBPγ, GATA-1, GATA-2 in HL-60 differentiation group were 20%, 21% and 18% respectively, which were significantly lower than those in untreated group. It is concluded that dysregulation of transcription factors is a key contributing factor in the pathogenesis of acute myeloid leukemia.
Cell Differentiation ; drug effects ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; RNA, Messenger ; genetics ; Transcription Factors ; metabolism ; Tretinoin ; pharmacology