China ; epidemiology ; Cross Infection ; epidemiology ; Hospitals ; Humans

OBJECTIVETo investigate changes in the adherent ability, the expression of adhesion related proteins Pyk2 and paxillin during HL-60 cells differentiation into granulocyte-monocyte induced by low-dose (LD) bufalin in combination with all-trans retinoic acid (ATRA), and to explore the effects of bortezomib on cellular adhesion and the expression of Pyk2 and paxillin.

METHODSThe expression of CD11b was detected by flow cytometry, cellular adherence ability by MTT assay, and the expressions of Pyk2, paxillin and tubulin by Western blot.

RESULTSThe combination of 5 nmol/L bufalin and 30 nmol/L ATRA induced HL-60 cells differentiation in a time-dependent manner, the percentages of CD11b positive cells treated for 2 d and 4 d being (20.0 +/- 2.8)% and (75.0 +/- 5.3)%, respectively, with the increasing of cellular adherence ability. Meanwhile the expressions of Pyk2 and Paxillin were also up-regulated in a time-dependent manner. Bortezomib suppressed HL-60 cell adhesion in a dose-dependent manner. At concentrations of 1 nmol/L and 10 nmol/L the adherence level were (7.8 +/- 0.1)% and (5.3 +/- 0.3)%, respectively, with down-regulation of Pyk2 but not Paxillin.

CONCLUSIONPyk2 is involved in the regulation of cellular adherence function. Bortezomib might inhibit HL-60 cells adhension function by down-regulation of Pyk2 expression.

Boronic Acids ; pharmacology ; Bortezomib ; Bufanolides ; pharmacology ; Cell Adhesion ; drug effects ; Cell Proliferation ; drug effects ; Focal Adhesion Kinase 2 ; metabolism ; HL-60 Cells ; Humans ; Paxillin ; metabolism ; Pyrazines ; pharmacology ; Tretinoin ; pharmacology

ObjectiveTo analyze Echinococcus infection in definitive and intermediate hosts in different zones of Qinghai plateau,Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,and to provideascientificbasisfor developing controlstrategiesagainstEchinococcosisinfection. Methods Echinococcosis infection in definitive hosts,dogs and foxes,was identified by morphological observation; in domesticated and wild intermediate host animals was identified by anatomy and pathology; some of the suspected samples were further identified by molecular biological methods.ResultsStray dogs in different zones of Qinghai plateau were infected with Echinococcus granulosus,the infection rates were 38.71％(300/775),49.60％(124/250),and 9.76％(4/41 ) in Qinghai southem plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant(x2 =25.72,P ＜ 0.01 ).in addition,only Qinghai southern plateau dogs were infected with Echinococcus multiloularis,and the infection rate was 16.04％(98/611).The infection rates of fox with Echinococcus multilocularis were 22.89％(38/166) and 30.77％(12/39) in Qinghai southern plateau and Qilian mountain-Hehuang valley,respectively,and wolves were also found to be infected with Echinococcus granulosus in the same areas.The infection rates of domesticated sheep,yaks,goats and pigs with Echinococcosis were significantly different statistically in those different areas(x2 =82.70,41.82,212.63,194.58,all P ＜ 0.01 ).The infection rates of sheep and yaks were higher[43.43％(5664/13 042),49.47％(2917/5896),52.99％ (887/1674),42.18％ (779/1847),50.70％ (1049/2069),52.90％ (685/1295) ] in three areas.The infection rates of goats and pigs [3.26％ (7/215),0.00％ (0/108)] in Qinghai southern plateau were lower than that of other two areas[ 19.51％(119/610),26.91％(43/1598),47.91％(343/716),21.91％(71/324)].The infection rates of Ochotona curzoniae with Echinococcosis were 6.21％ (243/3910),1.80％ (3/167) and 0.00％ (0/199) in Qinghai southern plateau,Qilian mountain-Hehuang valley and Chaidamu basin,respectively,and the difference was statistically significant (x2 =18.50,P ＜ 0.01 ).Moreover,wild intermediate hosts of Echinococcosis,such as Microtus fuscus,Lepus oiostolus,Pseudois nayaur,Procapra picticaudata,and Prodorcas gutturosa were found to be infected only in Qinghai southern plateau.ConclusionsHuman is faced with a threat of Echinococcosis infection from various definitive hosts in different zones of Qinghai plateau.And stray dogs are the most crucial factor.The life-cycles of Echinococcus are very complicated in Qinghai plateau.Qinghai plateau is a key area in prevention and control of Echinococcosis infection in China.

Antibody against the angiotensin AT1 receptor (AT1-Ab) could disturb placental development. The placenta is the key organ between mother and fetus. Placental damage will seriously impair fetal growth and development in utero, leading to intrauterine growth restriction (IUGR). Based on the fetal origins of adult disease (FOAD) hypothesis, IUGR could increase a propensity to develop adult onset cardiovascular disease (CVD). The present study was designed to determine whether vascular function has changed in the adult offspring of AT1-Ab positive pregnant rats. Twenty four female rats (8-week-old, AT1-Ab negative) were randomly divided into two groups, immunized and vehicle groups. Immunized group received active immunization to establish AT1-Ab-positive model, while vehicle group was subjected to Freund's adjuvant without antigen. After 8 weeks of immunization, the antibody titers in sera from the female rats were detected by enzyme-linked immunosorbent assay (ELISA). Then all the female rats were mated with normal Wistar male rats and became pregnant. Immunized/vehicle group offspring rats (I offspring/V offspring) were raised to 40-week-old under standard chow feeding. Then the two groups' offspring rats were given a high-salt diet for 12 weeks (4% NaCl in chow feeding). Systolic blood pressure (SBP) was measured dynamically by noninvasive blood pressure system. The vascular ring experiment was performed to detect vascular function and reactivity. As detected by ELISA, the titers of antibody peaked at the 8th week (OD values: 2.75 ± 0.08 vs 0.33 ± 0.01, P < 0.01 vs vehicle group at the same time point). There was no significant difference of SBP between the two groups' offspring rats during the high-salt diet (P > 0.05). Isolated thoracic aortic rings of I offspring had significantly decreased constriction under norepinephrine treatment (P < 0.01 vs V offspring) and significantly decreased dilation under acetylcholine treatment (P < 0.05 vs V offspring). These results suggest that the offspring of AT1-Ab-positive pregnant rats are more susceptible to vascular functional abnormality while being fed high-salt diet.
Animals ; Antibodies ; blood ; Cardiovascular Diseases ; etiology ; physiopathology ; Disease Susceptibility ; Female ; Fetal Growth Retardation ; physiopathology ; Immunization ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Wistar ; Receptor, Angiotensin, Type 1 ; immunology ; Sodium Chloride, Dietary ; administration & dosage ; adverse effects

Objective To evaluate the value of positron emission tomography(PET)-CT imaging combined with continual detection of CA_(125)in serum for diagnosis of early recurrent ovarian epithelial carcinoma.Methods Twenty six patients received PET-CT imaging,who were all diagnosed as primary epithelial ovarian cancer of stage Ⅱ-Ⅳ and had complete remission after cytoreductive surgery and multiple courses of chemotherapy in Shandong Provincial Cancer Hospital.After a steady period,all patients experienced progressive rising of CA_(125)values 3 times in 2 months.But no positive lesion was found by CT, or although suspicious positive focus was found,the recurrent and(or)metastatic extent was not definite. Out of 26 patients,16 were delivered rechemotherapy and(or)radiotherapy,and 10 received re- cytoreductive surgery.Results(1)Of 26 patients,the value of CA_(125)was more than 35 kU/L in 17,and in 14 of 17,pelvic or abdominal cavity recurrence was diagnosed by CT and PET-CT,and 4 showed simuhaneously distant metastasis on PET-CT.In the remaining 3 patients of which CT findings were negative,2 had pelvic and abdominal cavity recurrence,and one had bone metastasis on PET-CT.Of 9 patients with progressive rising CA_(125)levels but the value was less than cut-off(

In order to separate and purify the PEGylated recombinant human granulocyte stimulating factor (rhG-CSF) at large laboratory-scale level, a two-step ion-exchange chromatographic separation procedure was designed. Cation-exchange chromatography was applied first to separate PEGylated rhG-CSF from un-reacted rhG-CSF, followed by anion-exchange chromatography to dissolve individual PEG-rhG-CSF species (mono-, di- and tri-PEGylated rhG-CSF) and remove the free PEG. The molecular weight of individual PEGylated rhG-CSF was determined by MALDI-TOF and SDS-PAGE. MALDI-TOF mass spectrometry revealed that the molecular weights of mono-, di- and tri-PEGylated rhG-CSF are 23.8 kD, 28.6kD and 33.8kD, respectively. Cell proliferation activity was detected by MTT assay using NFS-60 cell. The in vitro residual bioactivity of mono-, di- and tri-PEGylated rhG-CSF were 90%, 75% and 43% respectively, comparing with the un-conjugated rhG-CSF. These results indicated that the un-conjugated rhG-CSF and excess free PEG can be removed completely and the three conjugate species can be purified into homogeneity by the two consecutive ion-exchange chromatographic steps. The purification procedure is easy to scale-up, high in performance and recovery.
Chromatography, Ion Exchange ; methods ; Granulocyte Colony-Stimulating Factor ; biosynthesis ; chemistry ; isolation & purification ; Humans ; Polyethylene Glycols ; chemistry ; isolation & purification ; Recombinant Proteins

To develop a sensitive and specific high performance liquid chromatography-tandem mass spectrometric (HPLC-MS/MS) method for the determination of mosapride in human plasma, mosapride and internal standard tamsulosin were extracted from plasma with liquid-liquid extraction, then separated on a Waters ACQUITY UPLC BEH C18 column (50 mm x 2.1 mm, 1.7 microm ID) with gradient elution at flow-rate of 0.25 mL x min(-1). The mobile phase was water (containing 0.3% formic acid) and acetonitrile under gradient conditions. Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 422 --> m/z 198 and m/z 409 --> m/z 228 were used to quantify mosapride and the internal standard, respectively. The linear calibration curve was obtained in the concentration range of 0.17 - 68.00 ng x mL(-1). The lower limit of quantification was 0.17 ng x mL(-1). The inter- and intra-day precision (RSD) was less than 13%, and the accuracy (RE) was within +/- 6.3% calculated from QC samples. The method was used to determine the concentration of mosapride in plasma after a single oral dose of 5 mg mosapride citrate to 20 healthy male Chinese volunteers. The method has been proved to be selective, sensitive, rapid and suitable for pharmacokinetic study of mosapride.
Administration, Oral ; Area Under Curve ; Benzamides ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Gastrointestinal Agents ; administration & dosage ; blood ; pharmacokinetics ; Humans ; Male ; Morpholines ; administration & dosage ; blood ; pharmacokinetics ; Sensitivity and Specificity ; Serotonin Receptor Agonists ; administration & dosage ; blood ; pharmacokinetics ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry ; methods

Objective To observe the inhibitory effect of CDglyTK double suicide gene on the growth of C6 glioma in vivo. Methods Thirty 4- to 6-week-old male Balb/c nude mice with subcutaneous injection of C6 glioma cells were randomized into two groups for injections with 5x105 CFU of CDglyTK (treatment group) or 50 μL PBS (control group) for 5 consecutive days. RT-PCR was performed to examine the mRNA expression of the CDglyTK fusion gene in the tumor cells. All the mice received intraperitoneal injections of 5-FC (500 mg/kg) and GCV (60 mg/kg) for 7 days, and the tumor volume, weight and mice survival time were observed. The effect of CDglyTK gene transfer on the apoptosis of the C6 glioma cells was evaluated using flow cytometry. Results RT-PCR demonstrated effective expression of the fusion gene in C6 glioma cells. In the fourth week of tumor cell inoculation, the tumor in the control group grew to the volume of 20 mmx30 mm with occasional death of the mice, and in week 6, all the control mice died. In the treatment group, the tumors showed no obvious growth with a volume of around 5 mm, and some of tumors even disappeared; no death of the mice occurred at the end of the third month. By gross observation, the tumors in the control mice were large and red with rich blood supply, but those in the treatment group showed reduced volume. On day 21 following tumor cell inoculation, the weight of the tumors was significantly greater in the control group than in the treatment group (2.51±0.58 vs 0.35±0.26 g, P<0.05), and the growth inhibition rate was 86.1% in the latter group. Flow cytometry showed significantly higher apoptotic rate of the tumor cells in the treatment group than in the control group (34.41%±5.2% vs 2.92%±1.3%, P<0.05), and electron microscope demonstrated the formation of apoptotic bodies in the tumor cells in the former group. Conclusion Significant antitumor effects can be obtained with the CKglyTK fusion gene in combination with the two prodrugs.

OBJECTIVETo analyze the effect and safety of bronchoscopic cryosurgery in children with lower airway stenosis caused by granulation and fiber hyperplasia.

METHODTwenty-two patients had undergone bronchoscopic examination and cryosurgery, 18 patients with atelectasis caused by Mycoplasma pneumoniae pneumonia, 2 patients with endobronchial tuberculosis (EBTB), 1 patient with atelectasis caused by granulation tissue after bronchial foreign body, and 1 case who had severe dyspnea, with tracheal stenosis after long-term endotracheal intubation during surgery for heart disease. All the patients under went bronchoscopic cryosurgery for several times, 1 case with severe tracheal stenosis was operated by electric coagulation before cryosurgery. Before and after the treatment, all the patients were evaluated based on clinical symptoms, tracheal lumen by bronchoscopy, chest CT for atelectasis and pulmonary function for tracheal stenosis.

RESULTThe patients were treated with cryosurgery for 1-4 times. Sixteen cases were markedly improved, clinical symptoms disappeared completely, no granulation and fiber hyperplasia were found during bronchoscopy, and all the atelectasis were cured. After the treatment, the bronchial lumen diameter of the patient with tracheal stenosis was increased from 2 mm to about 5 mm, the tidal volume increased from 3.0 ml/kg to 8.8 ml/kg. Five cases were effectively improved, the clinical symptoms also disappeared, during bronchoscopy and bronchial alveolar lavage (BAL), no granulation and fiber hyperplasia were found, but some distal bronchial tubes were atresia or narrowed, the lumen was patent, the atelectasis recovered partially. In 1 case the treatment was ineffective, as the clinical symptoms and granulation disappeared, but because almost all distal bronchi were atresia or narrow, the lumen was not patent during BAL, there was no change on chest CT. The total effective rate was 21/22 (95.5%). No severe complications occurred during and after the procedures. All the patient were followed up for 1 - 12 months, no case had recurrence.

CONCLUSIONTreatment with bronchoscopic cryosurgery is a safe and effective methods to tracheal stenosis caused by granulation tissue and fiber hyperplasia in children.

Bronchial Diseases ; etiology ; surgery ; Bronchoscopy ; Child ; Child, Preschool ; Cicatrix ; complications ; Constriction, Pathologic ; Cryosurgery ; Female ; Granuloma ; complications ; Humans ; Lung Diseases ; etiology ; surgery ; Male ; Tracheal Stenosis ; etiology ; surgery

OBJECTIVETo evaluate the effect of bronchoalveolar lavage (BAL) through bronchoscopy in diagnosis and treatment of accidental inhalation of liquid and powdery foreign bodies in children.

METHODTwenty-one cases of accidental inhalation were classified into the following 4 groups according to the materials inhaled: 3 cases inhaled the lipidic material; 2 cases inhaled chemical material; 15 cases inhaled hydrosoluble materials; 1 case inhaled powdery material. Eighteen cases were hospitalized within 3 days after the accidents, which included 16 cases treated within 1 day in hospital. And 3 cases were hospitalized 3 days after the accident. After hospitalization, all patients were immediately treated with direct aspiration and lavage clearance method through bronchoscopy. The inhaled foreign materials were cleared as soon as possible, which reduced the absorption, stimulation and infection of respiratory mucosa. The therapeutic effect was evaluated by endoscopic findings, clinical manifestation, laboratory examination and chest imaging. At the same time, decision was made on the second BAL and the prognosis was estimated.

RESULTTwenty-one cases had dyspnea of certain degree. The bronchial mucous membrane of all the 21 cases showed hyperemia, edema and exudate attached. Lavage fluid looked like broth and contained much of the inhaled material. After two to five times treatment, 18 cases were cured (86%), but 3 cases died (14%). One of these 3 cases fell into an oil well, 1 into mosquito repellent solution, another into manure pit. The patients who could not survive had severe airway obstruction, which was difficult to clean and made the problem deteriorated progressively. One of the patients was sent to a hospital 3 days after the accident and another case developed fungal infection, pneumothorax and finally respiratory failure.

CONCLUSIONBAL through bronchoscopy is necessary for children who accidentally inhaled liquid and powdery materials. This treatment is safe and effective in diagnosis and treatment of such problems in children. The effect of treatment is influenced by types of inhaled material, the time of lavage clearance through bronchoscopy, and certain complications, such as infection etc.

Bronchoalveolar Lavage ; Bronchoscopy ; Child ; Child, Preschool ; Female ; Foreign Bodies ; therapy ; Humans ; Infant ; Inhalation Exposure ; Male ; Powders