Objective To investigate the method of directed differentiation dendritic cells from embryonic stem cells(ESC) and to amplify high purity DCS in vitro for immunity therapy.Methods E14 ESC line were generated ESC-derived dendritic cells(ES-DC) in complete medium further supplemented with granulocyte-macrophage colony-stimulating factor(GM-CSF) and interleukin-3(IL-3).ES-DCs was used flow cytometry to determine CD11c,CD80,CD86,MHC-Ⅱ cell surface phenotype. Lipopolysaccharide (LPS) were added to induce the ES-DCs matured. The matured ES-DCs was harvested 24 hours later to be identified with morphology, transmission electron microscopy, analyzed by flow cytometry and compared with the immatured ES-DCs phenotype. The antigen presenting was evaluated by mixed lymphocyte responses.Results The ES-DC had obviously dendritic processes under scanning electron microscope . The immature DCs express low level of CD11c(4.33±0.23)%,CD80 (7.62±0.19) %, CD86 (4.77±1.22) % and MHC-Ⅱ (9.68±0.15) %, but the mature DCs express higher lerve of CD11c(47.36±2.68)%,CD80 (74.4±1.47) %, CD86 (29.77±2.00) % and MHC-Ⅱ (87.56±2.75) %. MLR showed that ES-DCs could effectively stimulate lymphocyte to proliferate.Conclusion These results provide evidence that DCs can be generated from E14 ESC with GM-CSF and IL-3, express high level of CD11c,CD80, CD86, MHC-Ⅱ and can effectively stimulate lymphocyte to proliferate. ES cells may become new origin for DCs which provided the immunotherapy.

OBJECTIVETo explore the effect and feasibility of acupuncture in uterine involution after cesarean section.

METHODSSeventy cases of cesarean section in primipara were randomized into an observation group (33 cases) and a control group (37 cases). In the control group, the conventional treatment was adopted after cesarean section. In the observation group, on the basic treatment as the control group, acupuncture was applied at Sanyinjiao (SP 6), Hegu (LI 4), Qihai (CV 6) and Guanyuan (CV 4). In 2 h after operation, acupuncture started, once a day, continuously for five times. The daily height of uterine fundus, daily uterine fundus decreasing degree, postpartum blood loss, lochia duration and ultrasonic B test in 42 days of postpartum were compared between the two groups.

RESULTSThe therapeutic methods of the two groups all promoted uterine contraction. The postpartum height of uterine fundus in the observation group was lower than that in the control group (all P<0.05) and the uterine fundus decreasing degree was higher than that in the control group (all P<0.05). The blood loss in 2 h, 6 to 12 h and 24 to 48 h of postpartum was less than that in the control group (all P<0.05). The lochia duration in the observation group was shorter than that in the control group (P<0.05).

CONCLUSIONAcupuncture promotes uterine contraction, reduces postpartum blood loss and lochia duration and benefits uterine involution after cesarean section.

Acupuncture Therapy ; Adult ; Cesarean Section ; adverse effects ; Female ; Humans ; Postoperative Complications ; physiopathology ; prevention & control ; therapy ; Uterine Contraction ; Young Adult

Adult ; Aged ; Aged, 80 and over ; Carcinoma in Situ ; virology ; Carcinoma, Squamous Cell ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; Microarray Analysis ; methods ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; diagnosis ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Uterine Cervical Neoplasms ; virology

Objective To explore the modulatory effect of 5-HT2A receptors on the discharge activities of inspiratory neurons in medial region of nucleus retrofacialis of neonatal rats. Methods Experiments were performed in vitro brainstem slice preparations from neonatal rats. These preparations included the medial region of nucleus retrofacialis with the hypoglossal nerve rootlets retained. The rhythmic discharges of the inspiratory neurons and activities of the hypoglossal nerve rootlets were simultaneously recorded by using microelectrodes and suction electrodes, respectively. Roles of 5-HT2A receptors in modulation of the discharge activities of inspiratory neurons were investigated by administration of the 5-HT2A receptor agonist 1-(2,5-dimethoxy-4-iodopbenyl)-2-aminopropane (DOI), and its specific antagonist ketanserine dissolved in modified Kreb's solution for perfused slices. Results In DOI group, the inspiratory time (TI) was (0.864±0.07)s, expiratory time (TE) was (10.78±1.06)s, respiratory cycle (RC) was (11.79±1.64)s, integral amplitude (IA) was (357.98±37.21)(μV·s) and the peak discharge frequency (PF) was (37.83±3.66)Hz. In control group, they were (0.68±0.06)s, (13.89±2.14)s, (14.77±1.92)s, (273.57±24.39)(μV·s), and (29.92±4.50)Hz, there were significant differences between the 2 groups (Pa<0.01). In ketanserine group, TI was (0.55±0.07)s, TE and RC were (18.43±3.28)s and (20.17+2.91)s respectively, IA and PF were (214.37±33.52)(μV·s) and (22.17±3.92)Hz, there were significant differences between ketanserine group and DOI, control group (Pa<0.01). Conclusion 5-HT2A receptors take part in modulate the discharge activities of inspiratory neurons in neonatal rat brainstem slices.

Objective To explore the modulation roles of gamma-aminobutyric acid A(GABAA) receptor on respiratory rhythmical discharge activity in brainstem slice of neonatal rats.Methods Experiments were performed on in vitro neonatal rats brainstem slices.The slices containing the medial region of the nucleus retrofacialis(mNRF)with the hypoglossal nerve rootlets retained was prepared in which the respira-tory-related rhythmic discharge activity(RRDA)were recorded from hypoglossal nerve rootlets by suction electrode.The possible role of GABAA receptor on RRDA was investigated by administration of GABA and its antagonist bicuculline in modified Kreb′s solution(MKS),for perfusion of the brainstem slices.Eighteen slices positive of RRDA were randomly divided into three equal groups:the first group of slices was perfused with different concentrations(10,20,40,60 ?mol/L) GABA and the most effective concentration was selected;the second group of slices was perfused with 10 ?mol/L bicuculline;the third group of slices was perfused with 10 ?mol/L bicuculline plus 40 ?mol/L GABA.The RRDA in 3 groups were observed.Results GABA decreased RRDA,40 ?mol/L was the most effective concentration.It shortened inspiratory time(TI),decreased integral amplitude(IA),prolonged respiratory cycle(RC) and expiratory time(TE) respectively.By contrast,10 ?mol/L bicuculline produced opposite effects on RRDA,it prolonged TI,increased IA,shortened RC and TE respectively.40 ?mol/L GABA plus 10 ?mol/L bicuculline had no significant effects on RRDA.Conclusion GABA plays an important role in modulate RRDA of neonatal rats via GABAA receptor.

The present study was carried out to determine the role of histamine H(1) and H(2) receptors in the generation of basic respiratory rhythm. Neonatal (aged 0-3 d) Sprague-Dawley rats of either sex were used. The medulla oblongata slice containing the medial region of the nucleus retrofacialis (mNRF) and the hypoglossal nerve rootlets was prepared and the surgical procedure was performed in the modified Kreb's solution (MKS) with continuous carbogen (95% O(2) and 5% CO(2)), and ended in 3 min. Respiratory rhythmical discharge activity (RRDA) of the rootlets of hypoglossal nerve was recorded by suction electrode. Thirty medulla oblongata slice preparations were divided into 5 groups. In groups I, II and III, histamine (5 μmol/L), H(1) receptor specific antagonist pyrilamine (10 μmol/L) and H(2) receptor specific antagonist cimetidine (5 μmol/L) was added into the perfusion solution for 15 min separately. In group IV, after application of histamine for 15 min, additional pyrilamine was added into the perfusion for another 15 min. In group V, after application of histamine for 15 min, additional cimetidine was added into the perfusion for another 15 min. The discharges of the roots of hypoglossal nerve were recorded. Signals were amplified and band-pass filtered (100-3.3 kHz). Data were sampled (1-10 kHz) and stored in the computer via BL-420 biological signal processing system. Our results showed that histamine significantly decreased the respiratory cycle (RC) and expiratory time (TE), but changes of integral amplitude (IA) and inspiratory time (TI) were not statistically significant. Pyrilamine induced significant increases in RC and TE, but changes of TI and IA were not statistically significant. Cimetidine had no effects on RC, TE, TI and IA of RRDA. The effect of histamine on the respiratory rhythm was reversed by additional application of pyrilamine but not cimetidine. Taken together, with the results mentioned above, histamine H(1) receptors but not H(2) receptors may play an important role in the modulation of RRDA in the medulla oblongata slice preparation of neonatal rats.
Animals ; Animals, Newborn ; Cimetidine ; pharmacology ; Female ; Histamine ; pharmacology ; Histamine H1 Antagonists ; pharmacology ; Histamine H2 Antagonists ; pharmacology ; Hypoglossal Nerve ; physiology ; In Vitro Techniques ; Male ; Medulla Oblongata ; physiology ; Pyrilamine ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Histamine H1 ; physiology ; Receptors, Histamine H2 ; physiology ; Respiration

To investigate the effects of nikethamide on the generation and modulation of rhythmic respiration of neonatal rats and the role of 5-HT(2A) receptor in this course, experiments were performed on the transverse medullary slices of neonatal rats (both sexes, 1-3 d) in vitro. The slices containing the medial region of the nucleus retrofacialis (mNRF) with the hypoglossal nerve rootlets were prepared in which the respiratory-related rhythmic discharge activity (RRDA) was recorded from the hypoglossal nerve rootlets by suction electrode. The possible role of nikethamide on RRDA was investigated by administration of an agonist of 5-HT(2A) receptor, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI), and an antagonist of 5-HT(2A) receptor, ketanserine, dissolved in modified Krebos solution (MKS). Thirty slices were randomly divided into five groups: Group 1: the slices were perfused with different concentrations of nikethamide (0.5, 1, 3, 5, 7, 10 μg/mL), and the most effective concentration was selected; Group 2: the slices were perfused with DOI (40 μmol/L); Group 3: the slices were perfused with ketanserine (40 μmol/L); Group 4: the slices were perfused with ketanserine + DOI; Group 5: the slices were perfused with nikethamide, then perfused with nikethamide + ketanserine after washout of nikethamide. Nikethamide increased RRDA in transverse medullary slices at 0.5-7 μg/mL, and 5 μg/mL was the most effective concentration. DOI increased RRDA with prolonged inspiratory time (TI), increased integral amplitude (IA), and shortened respiratory cycle (RC). Ketanserine decreased RRDA with shortened TI, decreased IA and prolonged RC. Ketanserine + DOI had no significant effects on RRDA. The effects of nikethamide on RC and IA were totally and partially reversed by additional application of ketanserine, but the effect of nikethamide on TI was not influenced by ketanserine. It is proposed that nikethamide increases RRDA partly via 5-HT(2A) receptors.
Animals ; Animals, Newborn ; Female ; In Vitro Techniques ; Male ; Medulla Oblongata ; drug effects ; physiology ; Nikethamide ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptor, Serotonin, 5-HT2A ; metabolism ; Respiration ; drug effects ; Respiratory Center ; physiology ; Serotonin

Objective:To determine the half lethal dose(LD50) of paraquat in rats and to establish a relatively safe and stable pathological animals model of pulmonary fibrosis.Methods: Ninety-six SD rats totally in half genders.Fifty SD rats in half genders were randomly divided into 5 groups,each had 10 rats.Feed the rats with different doses of liquor of paraquat intraperitoneally one time and definite the half lethal dose of one and two weeks.After that,prepare another forty-six SD rats,also in half genders,as intonication group,twenty-eight rats were treated with the liquor of paraquat in dosage of 18 mg/kg intraperitoneally.As control group,sixteen rats were treated with equivalent volume of normal saline.Observe the toxic symptom daily and rats were sacrificed on day 1,3,5,7,14,21,28,35 and 42 respectively for the histological examination.Results: The half lethal doses of intraperitoneal paraquat of 1 and 2 weeks were 18.27 and 17.29,with 95% confidence intervals of 16.61-20.09 and 15.99-18.67,respectively.After intraperitoneal paraquat injection at the dose of 18 mg/kg,typical toxic symptoms were observed at different times in the rats.The whole process of acute lung injury and fibrosis induced by paraquat intoxication could be seen with the naked eyes or under the light microscope.Conclusion: Paraquat has a strong toxicity to rats.A proper dose of paraquat solution can not only reduce the number of experimental rats,but also induce typical pulmonary fibrosis in rats.

Objective To identify risk factors and preventive measures of nosoconial infections in patients with non-Hodgkin lymphoma (NHL). Methods Clinical data of 65 NHL patients admitted from January to December 2007 were retrospectively analyzed. Results According to WHO classification (2001), 58 patients were with B-cell lymphoma, 7 were with T-cell lymphorna. All patients received CHOP regimen as initial chemotherapy and 23 of them were with nosecomial infections. Logistic regression analysis demonstrated that age, length of stay, pathological type, bone marrow involvement, levels of serum lactate dehydrogenase (LDH), beta2-microglobulin and invasive treatment were identified as risk factors of nosocomial infections. Respiratory tract infections and infections with gram-negative microorganisms were the most popular. Conclusion High nosocomial infection rate is found in NHL patients, and control of risk factors may effectively prevent nosocomial infections in NHL patients.

OBJECTIVE:To establish a method for the content determination of matrine,sophocarpine,emodin,rhein,chloro-genic acid,sarkosaponin a,aloe-emodin in Kuhuang injection.METHODS:UPLC-MS/MS was adopted. The determination was per-formed on Phenomenex Kenetix C18 column with mobile phase consisted of methanol-0.1% formic acid(gradient elution)at the flow rate of 0.5 mL/min. The column temperature was set at 20 ℃,and injector temperature was set at 10 ℃. The equilibrium time was 4 min and sample size was 5 μL. Ionization mode was electrospray ionization with spray voltage of 5500 V and -4500 V,at-omizing air pressure of 4.14×105 Pa,heater pressure of 4.48×105 Pa,curtain air of 1.72×105 Pa and ion source temperature of 600 ℃. The work mode was multiple reaction monitoring mode. RESULTS:The linear range were 1.25-80.0 ng/mL for matrine (r=0.9993),1.10-70.0 ng/mL for sophocarpine(r=0.9995),0.16-10.5 ng/mL for emodin(r=0.9993),2.61-168 ng/mL for rhe-in (r=0.9993),1.50-96.0 ng/mL for chlorogenic acid (r=0.9993),1.48-94.5 ng/mL for sarkosaponin a (r=0.9996) and 6.11-391 ng/mL for aloe-emodin(r=0.9991). The limits of quantification were 0.061,0.109,0.041,1.313,0.500,0.492,3.055 ng/mL,limits of detection were 0.025,0.054,0.016,0.656,0.150,0.148,1.528 ng/mL. RSDs of precision,stability and reproducibility tests were all no more than 3%. The recoveries of them were 95.45%-99.45%(RSD=1.43%,n=6),97.50%-101.00%(RSD=1.50%,n=6),95.67%-101.73%(RSD=2.85%,n=6),97.17%-100.57%(RSD=1.16%,n=6),95.19%-98.90%(RSD=1.71%, n=6),95.38%-103.85%(RSD=3.39%,n=6),95.50%-101.17%(RSD=1.20%,n=6),respectively. CONCLUSIONS:The method is simple,effective,accurate,reliable and suitable for simultaneous determination for 7 components in Kuhuang injection.