Objective A HPLC method was established to measure the contents of major nucleosides in Hyriopsis cumingii.Method Separation was performed on a Lichrosper-C_(18) column, pure water was used as mobile phase,the flow rate was 1.0mL min~(-1),the column temperature at 30℃and the detective walvelength at 254 nm.Results The contents of uracil,hypoxanthine, xanthine,uridine and thymine were 153.2～173.3?g?g~(-1),106.6～360.8?g?g~(-1),117.6～172.8?g?g~(-1),70.3～153.1?g?g~(-1) and 343.9～511.4?g?g~(-1),respectively. Conclusion This method showed good repeatability and flexibility.It could be used as a conventional method for determination of major nucleosides in H.cumingii.

The dynamic expression of cytokines and phenotypes during the differentiation process of dendritic cell precursors (pre-DCs) to mature dendritic cells (DCs) was investigated,and the effects of inflammatory stimulation with lipopolysaccharide (LPS) on DCs differentiation were understood.The differentiation of bone marrow cells isolated from Balb/c mice was induced to DCs in an 8-day cell culture system with RPMI-1640 complete culture medium containing 10％ FBS,20 ng/mL recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and 10 ng/mL recombinant mouse interleukin-4 (rmIL-4).On the day 3,6 and 7 after culture,DCs were divided into non-LPS group and LPS group,given 500 ng/mL LPS for 24 h stimulation and no stimulation respectively.The expression levels of CD 1 l c+,MHC-Ⅱ +,CD40+,CD80+ and CD86+ were detected by flow cytometry,and those of IL-2,IL-4,IL-10,IL-12 p70 and IFN-γ in the supernatant by ELISA.On the day 3 and 6 after culture,the expression of IL-2,IL-4,IL-10 and IFN-γ in DCs showed no significant differences between non-LPS group and LPS group,whereas the differences were significant at day 7.The expression levels of cytokines (for IL-2,IL-4,IL-10,IFN-γ and IL-12 p70:152.86±6.91,778.33±8.35,44.55±2.54,5.8.26±1.09 and 2423.00±57.21 pg/mL respectively) in LPS group were higher than those in non-LPS group,especially IL-12 p70 increased obviously at day 7.It was concluded that during the differentiation process of pre-DCs to mature DCs,LPS stimulates DCs producing large amounts of IL-12 p70 and Thl-type cytokines as compared with Th2-type cytokines,and day 7 may be a key time-point for DCs polarization.

The aim of the present study was to explore the effects of two different modes of intermittent hypobaric hypoxia (IHH) on myocardial ischemia/reperfusion injury in developing rat hearts. Postnatal male sprague-Dawley rats (n=72) were divided randomly into 3 groups: intermittent hypoxia at 3 000 m (IHH3000) group, intermittent hypoxia at 5 000 m (IHH5000) group and control group. The isolated hearts were perfused in the Langendorff apparatus, undergoing 30 min of global ischemia and 60 min of reperfusion. Cardiac function, coronary flow and lactate dehydrogenase (LDH) activity were recorded at 5 min before ischemia and 1, 5, 10, 20, 30, 60 min during reperfusion, respectively. The heart weight was measured at the end of the experiment. The results showed that: (1) There was no difference in body weight gaining between IHH3000 and control groups. The gain of body weight in IHH5000 group was much lower than that in IHH3000 and control groups (P<0.01). (2) Compared with that in the control group, the recovery of cardiac function in IHH3000 group was enhanced at 60 min after ischemia/reperfusion, coronary flow was increased, and LDH activity was decreased (P<0.05), meaning a cardioprotective effect occurred. There was no significant difference in heart weight between IHH3000 and control groups. In addition, cardiac function restored better in IHH3000 group after 42 d of hypoxic exposure than that after 28 d of hypoxic exposure (P<0.05). (3) Compared with that in the control group, the recovery of cardiac function in IHH5000 group was lower, coronary flow was decreased, and LDH activity was increased (P<0.05). There was a hypertrophy in the right ventricle in IHH5000 group. All changes indicated definitely that a detrimental effect developed in IHH5000 group. The results suggest that proper IHH can protect developing rat hearts against ischemia/reperfusion injury while this effect could be affected by the modes of intermittent hypoxic exposure.
Animals ; Heart ; Heart Ventricles ; Hypoxia ; Male ; Myocardial Reperfusion Injury ; prevention & control ; Protective Agents ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury

AIMTo elucidate the effect of CIHH on cellular immunity and humoral immunity in rat by using flow cytometry method, immunohistochemistry method and electron microscopy techniques.

METHODSForty-eight male adult Sprague-Dawley rats were randomly divided into 4 groups: control(CON) group, 14 days CIHH (CIHH14) group, 28 days CIHH (CIHH28) group, 42 days CIHH (CIHH42) group. The animals in CIHH groups were exposed to 14, 28 and 42 days hypobaric hypoxia(simulated 3 000 m altitude, 5 h per day), respectively. Half of the animals in each group was treated with normaxia and the other half animals were treated with acute hypoxia for 1 h. CD3, CD4, CD8 T lymphocytes, natural killer (NK) cells, IgG, cortisol, epirenamine and C-reactive protein were examined. The weight and ultrastructure of thymus and spleen were observed.

RESULTS(1) Compared with CON, both indexes of thymus and spleen in CIHH14 rats were increased significantly. Spleen index, but not thymus index, was increased in CIHH28 and CIHH42 rats. The thymocytes and spleen cytes in rat were injuryed during acute hypoxia, but the damage in CIHH rats was significant slighter than that in CON rats. (2) Compared with CON, CIHH28 and CIHH42, CD8 in CIHH14 rats were decreased, ratios of CD4/CD8 was increased and NK was decreased. (3) The rats of CON during acute hypoxia showed that CD4 was increased, CD8 was decreased, ratio of CD4/CD8 was elevated, and NK was increased. But there were no significant changes of CD3, CD4, CD8 and NK in CIHH28 and CIHH42 animals during acute hypoxia. (4) Compared with CON, CIHH28 and CIHH42, cortisol in CIHH14 rats was increased obviously, Epirenamine, cortisol and C-reactive protein in CON rats were increased, but there were no obvious changes in CIHH rats before and after acute hypoxia.

CONCLUSIONCIHH protects the immune function of rat against acute hypoxia, which is related with the regulation of neuroendocrine.

Altitude Sickness ; physiopathology ; Animals ; Atmospheric Pressure ; Hypoxia ; physiopathology ; Immunity, Cellular ; physiology ; Immunity, Humoral ; physiology ; Male ; Neuroimmunomodulation ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spleen ; immunology ; T-Lymphocytes ; immunology ; Thymus Gland ; immunology

OBJECTIVETo compare the long-term efficacy of procedure for prolapse and hemorrhoids (PPH) and Milligan-Morgan hemorrhoidectomy (MMH) in the treatment of III and IV degree internal hemorrhoids.

METHODSOne hundred patients were randomly divided into two groups and received PPH (n=42) and MMH (n=58) respectively. After two years, the efficacy, complications and function of defecation were compared.

RESULTSTwo years after operation, the morbidities of hydrorrhea (2.38% vs 20.69%, P=0.007), dermal neoplasm formation (9.52% vs 25.86%, P=0.040) and narrowing in the caliber of the stools (2.38% vs 18.97%, P=0.027) were significantly lower in PPH group than those in MMH group (P<0.05). The morbidities of overall complication (9.52% vs 25.86%, P=0.040) and overall abnormal function of defecation (9.52% vs 29.31%, P=0.017) were lower in PPH group than those in MMH group (P<0.05). However, there were no significant differences of the morbidity of relapse (14.29% vs 10.34%, P=0.549), patient satisfactory degree (92.86% vs 87.93%, P=0.636) and overall symptom recurrence rate (19.05% vs 25.86%, P=0.424) between the two groups.

CONCLUSIONSLong-term efficacies of procedure for prolapse and hemorrhoids and Milligan-Morgan haemorrhoidectomy in the treatment of III and IV degree internal hemorrhoids are similar. PPH has better safety, less complications and less effect on abnormal function of defecation compared with MMH.

Adolescent ; Adult ; Aged ; Female ; Hemorrhoids ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Mucous Membrane ; surgery ; Prolapse ; Sutures ; Treatment Outcome ; Young Adult

Adult ; Carcinoma ; virology ; Cervical Intraepithelial Neoplasia ; virology ; Female ; Human papillomavirus 16 ; isolation & purification ; Human papillomavirus 18 ; isolation & purification ; Humans ; In Situ Hybridization ; Middle Aged ; Papillomavirus Infections ; Uterine Cervical Neoplasms ; virology

The purpose of the present study was to investigate the effect of chronic intermittent hypobaric hypoxia (CIHH) on α(1)-adrenergic receptors and the role of alpha(1)-adrenergic receptors in the protection of CIHH against ischemic injury of myocardium. Sixty-six adult male Sprague-Dawley rats were randomly divided into four groups: control group (Con), 14-day CIHH treatment group (CIHH14), 28-day CIHH treatment group (CIHH28) and 42-day CIHH treatment group (CIHH42). CIHH rats were exposed to hypoxia mimicking 5 000 m altitude (p(B)=404 mmHg, p(O(2))=84 mmHg) in a hypobaric chamber, 6 h daily for 14, 28 and 42 d, respectively. Control animals lived in the same environment as CIHH animals except hypoxia exposure. After anesthesia with sodium pentobarbital (3.0-3.5 mL/kg body weight, i.p.), papillary muscle was taken from the right ventricle of rat and perfused with modified Tyrode's solution continuously, at constant temperature (37 °C) and perfusion speed (12 mL/min). Muscle contraction was evoked by electric stimuli. Different concentrations (1x10(-7), 1x10(-6) and 1x10(-5) mol/L) of phenylephrine (PE), an alpha(1)-adrenergic receptor agonist, were applied cumulatively to investigate the effect of PE on the mechanic contraction of right ventricular papillary muscles of rats in Con, CIHH14, CIHH28 and CIHH42 groups. Also, prazosin (1x10(-6) mol/L), an α(1)-adrenergic receptor antagonist, was used to investigate the role of α(1)-adrenergic receptor in the protective effect of CIHH on papillary muscle. The results showed: (1) PE increased the maximal isometric tension (P(max)) and maximal velocity of tension development (P(dT/dt)) of muscle contraction in a dose-dependent manner (P<0.05), and the increase of the muscle contraction was much greater in CIHH28 and CIHH42 rats than that in Con rats (P<0.05). Under 1x10(-5) mol/L of PE, the increases of P(max) and P(dT/dt) over the baseline were 51.2% and 44.5% in CIHH28 group, 48.6% and 44.5% in CIHH42 group, and 28.7% and 24.5% in Con group, respectively; (2) The contraction of papillary muscle decreased during simulated ischemia, but the decrease was slighter in CIHH rats than that in Con rats (P<0.05). The decreases in P(max) and P(dT/dt) were 59.6% and 53.6% in CIHH28 group, 60.4% and 49.9% in CIHH42 group, and 74.4% and 64.7% in Con group, respectively; (3) The protective effect of CIHH on ischemic papillary muscle was abolished by prazosin (1x10(-6) mol/L). The results of the present study suggest that CIHH increases the activity of α(1)-adrenergic receptor, which is possibly one of the mechanisms for the cardioprotection of CIHH.
Altitude ; Animals ; Heart Ventricles ; physiopathology ; Hypoxia ; metabolism ; Male ; Muscle Contraction ; Myocardium ; metabolism ; Papillary Muscles ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Adrenergic, alpha-1 ; metabolism

Objective To evaluate uncertainty for the determination of oxcarbazepine active metabolite (10-hydroxycarbazepine) in human se-rum by solid phase extraction -HPLC.Methods The uncertainty caused in the whole process of determination was analyzed.Sources of uncertainty were established and simplified, then each component of un-certainty, combined uncertainty and expanded uncertainty were calculat-ed by using the method validation data in accordance with the evaluation procedure.Results The expanded uncertainty for low, medium and high concentrations (3.18, 12.73, 50.91 μg· mL-1 ) of 10-hydroxy-carbazepine was 0.24, 0.44 and 1.66 μg· mL-1 , respectively ( k =2, P=95%).Conclusion The uncertainty of this method was mainly caused by calibration curve fitting ( especially low level of sample ) , serum drug extraction, serum sample preparation, HPLC error and measuring repeatability.

OBJECTIVETo identify the clinical and pathological features of acute myeloid leukemia with B lymphoproliferative disorders.

METHODSThe characteristics of 3 cases of acute monocytic leukemia with untreated chronic lymphocytic leukemia/monoclonal B-cell lymphocytosis were reported with literatures review.

RESULTSThe patients presented with a history of anemia, bleeding and/or fever. Acute monocytic leukemia was diagnosed by bone marrow morphology, cytochemistry and pathology studies. Immunophenotyping by flow cytometry analysis showed a significant population of absolute B-lymphocyte count of > 5×10(9)/L in a patients, similar to that of chronic lymphocytic leukemia.

CONCLUSIONSThe association of acute monocytic leukemia and untreated chronic lymphocytic leukemia/monoclonal B-cell lymphocytosis was a rare event. The abnormal B lymphocytes was likely to be misdiagnosis. Thus, it was important to combine several kinds of laboratory studies, especially flow cytometry to identify this rare disorder.

Aged ; B-Lymphocytes ; pathology ; Female ; Humans ; Leukemia, Monocytic, Acute ; complications ; diagnosis ; pathology ; Lymphocytosis ; complications ; diagnosis ; pathology ; Middle Aged

OBJECTIVETo investigate The modulating role of p38 mitogen-activated protein kinase (MAPK) in the expression of tumor necrosis factor-alpha in hepatic cells and its role in hepatic injury in severely burned rats.

METHODSTwenty-four adult healthy male SD rats were randomly divided into three groups (8 rats in each group): sham group, burn group, and burn with SB203580 group. A rat model of full-thickness burn injury covering 30% total body surface area (TBSA) was reproduced. The specific inhibitor of p38MAPK (SB203580 in 10 mg/kg) was given to the rats in the burn with SB203580 group at 15 minutes and 12 hours after burn. The serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured at 24 postburn hours (PBHs). The TNF-alpha mRNA expression in the liver was determined by real-time reverse transcription polymerase chain reaction, and the expression levels of p38MAPK and phosphor-p38MAPK in the liver were determined by Western blot analysis.

RESULTSThe serum levels of AST and ALT, and the expression of TNF-alpha mRNA in liver cells were significantly higher in burn group than those in sham and SB203580 groups (P < 0.05 or 0.01), but there was no difference between the two latter groups. It was indicated by Western blot results that there was no difference of p38MAPK expression in rat liver among the three groups (P > 0.05). The phospho-p38MAPK expression ratio among sham, burn and burn with SB203580 groups was 1.00:3.90:1.10. The phospho-p38MAPK expression was significantly lower in burn with SB203580 group than that in burn group (P < 0.01), but there was no significant difference compared with that in sham group (P > 0.05).

CONCLUSIONThe postburn activated p38MAPK in rat liver after severe burn injury enhances the expression of TNF-alpha mRNA and participates in the development of postburn hepatic injury.

Animals ; Blotting, Western ; Burns ; metabolism ; pathology ; Liver ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism