Objective To research the effect of ulinastatin on T-cytoimmunity in patients with infertility undergoing laparoscopic surgery. Methods Forty patients scheduled for receiving laparoscopic surgery were equally randomized into two groups, ulinastatin group (Group U) and control group (Group C). Ulinastatin was given to patients in the Group U at a dose of 20 × 104 U before anesthetic. No ulinastatin was given to patients in the Group C. Patients′venous blood samples for T-lymphocyte subset (CD3+,CD3+CD4+,CD3+CD8+) and CD3+CD4+/ CD3+CD8+ ratio calculation were collected before the surgery (T0) and at 0 h (T1),1st day (T2),3rd day (T3) after the surgery. Results CD3+ had less difference at T1~3 compared with T0 in the Group C but raised obviously at T2~3 in the Group U. CD3+CD4+ were only raised at T3 compared with T0 but raised obviously at T2~3 in the Group U. CD3+CD8+ were raised obviously at T2~3 compared with T0 in the Group C but had less difference in the Group U. CD3+CD4+/CD3+CD8+ had less difference at T1~3 compared with T0 in the Group C but raised obviously at T3 in the Group U. Conclusion The application of ulinastatin in laparoscopic surgery could significantly produce protective effect on T-cytoimmunity.

An endophytic bacterium strain EBS05 from Cinamonum camphra was identified as Bacillus subtilis by morphological taxonomy and sequence analysis of 16S~23S rRNA intergenic spacer regions. Properties of antimicrobial compound produced by EBS05 were assayed. The active compound had the maximum absorbance peak at ?213.5 nm. The antimicrobial activity was stable in solution with pH value from 5 to 8, and decreased significantly in solution with pH value less than 4.0 or more than 9.0. The antimicrobial compound had thermodynamics stability. Its activity changed a little after treated at 60?C~80?C for two hours, and compared with 65% original activity after treated at 1?105 Pa for 30 minutes. The active substance had high resistance to ultraviolet radiation and protease K. Antimicrobial compound was soluble in alcohol solu- tion, which was easily dissolved in methanol and ethanol, but not dissolved in ethyl acetate, acetonitrile and petroleum et al.

In order to explore the clinical hypolipidemic features of Daidai flavone extract, the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats were studied and compared. The study established the quantitative determination method of naringin and neohesperidin in plasma by UPLC-MS. Study compared the pharmacokinetics differences of naringin and noehesperidin in normal and hyperlipemia rats on the basis of establishment of hyperlipemia model. Results indicated that the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats showed significant differences. The C(max) of naringin and neohesperidin in hyperlipemia rats plasma after oral administration of Daidai flavone extract increased obviously, while t1/2, MRT and AUC0-24 h decreased, compared to normal rats. But t(max) showed no differences to that of normal rats. The results further proved Daidai flavone extract would have better hypolipidemic effect in the hyperlipemia pathological status. And the characteristic active ingredients naringin and noehesperidin were the material base of Daidai flavone extract to express the hypolipidemic effect.
Animals ; Citrus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Flavones ; chemistry ; Hyperlipidemias ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the regulatory effect of Ligustrazine Injection (LI) on the cellular immune function in patients undergoing autologous blood transfusion (ABT).

METHODSEnrolled were 60 patients scheduled for receiving selective lumbar surgery at the Department of Spinal Orthopedics, First Hospital Affiliated to Guangzhou University of Traditional Chinese Medicine during October 2009 to June 2010. They were equally randomized into two groups, the trial group and the control group. LI was given to patients in the trial group by intravenous dripping at the dose of 2 mg/kg 30 min before autologous blood collection. The LI (at the final concentration of 0.005%) was added in the heparin saline solution and the washing saline for recycle blood. No LI was given to patients in the control group. They received the same treatment of the trial group. The operation time, the amount of blood loss and blood transfusion were recorded. Patients' venous blood samples were collected for determining cytokines including interleukin-2 (IL-2), interleukin-10 (IL-10), interferon-gamma (IFN-gamma) by ELISA and calculating IL-2/IL-10 ratio before surgery (T1), 1 h (T2), 1 day (T3), and 5 days (T4) after ABT.

RESULTSThere was no statistical difference in the amount of blood loss and blood transfusion, the levels of IL-2, IL-10, IFN-gamma, or IL-2/IL-10 at T1 between the two groups (P>0.05). Compared with T1 of the same group, the level of IL-2 decreased at T(2-4), IL-10 increased and IL-2/IL-10 decreased at T(2-3) in the two groups. The level of IFN-gamma decreased at T(2-4), IL-2/IL-10 increased at T4, the level of IL-10 decreased at T4 in the control group (P<0.05, P<0.01). The level of IL-10 decreased at T4 in the trial group with statistical difference (P<0.05, P<0.01). Compared with the control group, the level of IL-2, IFN-gamma, and IL-2/IL-10 at T(2-4) were obviously higher in the trial group. But the IL-10 level was lower in the trial group than in the control group at T(2-4) (P<0.05, P<0.01).

CONCLUSIONThe application of LI in ABT had regulatory effects on the balance of cytokines.

Adult ; Aged ; Blood Transfusion, Autologous ; Female ; Humans ; Immunity, Cellular ; drug effects ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-2 ; blood ; Male ; Middle Aged ; Postoperative Period ; Pyrazines ; therapeutic use ; Spine ; surgery ; Young Adult

In the treatment of hypertensive crisis, the novel Rho kinase inhibitor DL0805-2 can rapidly lower systematic blood pressure, reduce pulmonary artery pressure, and has a significant protective effect on lung injury. This experiment intends to evaluate the efficacy of DL0805-2 against pulmonary arterial hypertension (PAH) and preliminarily reveals its underlying mechanism. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences. Sprague Dawley (SD) rats were randomly divided into DL0805-2 low, medium, and high dose groups (1, 3, and 10 mg·kg^-1), bosentan positive control group, model group, and blank control group. The drug was administered daily on the 7th day after model establishment by monocrotaline injection. On the 25th day of the experiment, relevant indicators were examined to observe the therapeutic effect of DL0805-2 on pulmonary hypertension. DL0805-2 significantly relieved the abnormal changes in the physiological parameters related to PAH induced by monocrotaline, including reducing right ventricular systolic pressure, alleviating cardiac damage caused by pressure overload, and reducing the levels of endothelin-1 and inflammatory factors in lung tissues. DL0805-2 also attenuated pulmonary arteries remodeling. It was preliminarily discovered that DL0805-2 exerts preventive and therapeutic effect on PAH through Rho-kinase pathway. Our results suggested that DL0805-2 had good therapeutic effects on monocrotaline-induced PAH rat model. It intervened early in the disease process, effectively prevented the development of the disease, and reduced the mortality of the diseased animals. The mechanism is related to Rho-kinase pathway.

AIM To establish an HPLC method for the content determination of six constituents in Shiyifang Vinum (Notoginseng Radix et Rhizoma,Dipsaci Radix,Carthami Flos,etc.).METHODS The content determination of notoginsenoside R1,ginsenoside Rg1,ginsenoside Rb1 and asperosaponin Ⅵ was performed on a 30℃ thermostatic Inertsil(R) ODS-3 C18column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1.0 mL/min in a gradient manner,and the detection wavelength was set at 203 nm.The content determination of brucine and strychnine was conducted on a 30 ℃ thermostatic Geminni(R) C18 110(A) column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-mixed solution of 0.01 mol/L sodium heptanesulfonate and 0.02 mol/L potassium dihydrogen phosphate flowing at 1.0 mL/min in an isocratic elution manner,and the detection wavelength was set at 260 nm.RESULTS Six constituents showed good linear relationships within their own ranges (r ＞ 0.999 0),whose average recoveries were 98.52％-99.96％ with the RSDs of 2.0％-2.3％.CONCLUSION This simple,accurate and reproducible method can be used for the quality control of Shiyifang Vinum.

OBJECTIVETo explore the expression and sequence of human MutS homologue 2 (hMSH2) during different stages of human bronchial epithelial (16HBE) cells induced by cadmium chloride (CdCl2).

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical staining (SP method) were used to measure the hMSH2 mRNA and protein expression in 16HBE cells and its different passage cells treated by CdCl2 (the 5th, 15th, 35th passage, and neoplasm cells from nude mice's tumor tissue). hMSH2 exon 6, hMSH2 exon 7, hMSH2 exon 8, hMSH2 exon 9, hMSH2 exon 12 of the 16HBE cells and neoplasm cells from nude mice's tumor tissue were amplified by polymerase chain reactions (PCR). The amplified DNA strips were purified. Then the exons were detected by DNA analysis.

RESULTSDuring the passages of 16HBE cells treated with CdCl2, the expression of hMSH2 gene were decreased gradually. The hMSH2 gene mRNA and protein expression levels of the CdCl2 transformed 35th 16HBE cells and tumorigenic cells of nude mice significant decreased compared with non-transformed 16HBE cells (P < 0.01). In the tumorigenic cells of nude mice induced by CdCl2, there were thymine (T) deletion in 1st, 2nd and 7th site of hMSH2 exon 8, there were adenine (A) deletion in 20th and 182th site of hMSH2 exon 9, there were adenine (A) insertion in 241st site of hMSH2 exon 12. All the mutations were frame shift mutation.

CONCLUSIONThe expression decreased and the mutation of hMSH2 gene may be the possible carcinogenic mechanism for CdCl2.

Animals ; Bronchi ; cytology ; Cadmium Chloride ; toxicity ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; genetics ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Humans ; Mice ; Mice, Nude ; MutS Homolog 2 Protein ; genetics ; metabolism ; Mutation ; RNA, Messenger ; genetics

BACKGROUNDThe evoked electromyography (EMG) is frequently used to identify facial nerve in order to prevent its damage during surgeries. Partial neuromuscular blockade (NMB) has been suggested to favor EMG activity and insure patients' safety. The aim of this study was to determine an adequate level of NMB correspondent to sensible facial nerve identification by evaluating the relationship between facial EMG responses and peripheral NMB levels during the middle ear surgeries.

METHODSFacial nerve evoked EMG and NMB monitoring were performed simultaneously in 40 patients who underwent tympanoplasty. Facial electromyographic responses were recorded by insertion of needle electrodes into the orbicularis oris and orbicularis oculi muscles after electrical stimulation on facial nerve. The NMB was observed objectively with the hypothenar muscle's twitching after electrical stimulation of ulnar nerve, and the intensity of blockade was adjusted at levels of 0, 25%, 50%, 75%, 90%, and 100% respectively with increased intravenous infusion of Rocuronium (muscle relaxant).

RESULTSAll of the patients had detectable EMG responses at the levels of NMB or= 75%. A significant linear positive correlation was present between stimulation thresholds and NMB levels while a linear negative correlation was present between EMG amplitudes and NMB levels.

CONCLUSIONSThe facial nerve monitoring via facial electromyographic responses can be obtained when an intraoperative partial neuromuscular blockade is induced to provide an adequate immobilization of the patient. The 50% NMB should be considered as the choice of anesthetic management for facial nerve monitoring in otologic microsurgery based on the relationship of correlation.

Adult ; Ear, Middle ; surgery ; Electromyography ; methods ; Facial Nerve ; physiology ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Neuromuscular Blockade ; methods ; Otologic Surgical Procedures ; methods ; Young Adult

OBJECTIVETo evaluate the protective effect of ligustrazine on T-lymphocyte immunity of patients undergoing autologous blood transfusion (ABT).

METHODSForty American Society of Anesthesiologist (ASA) I - II patients scheduled for receiving selective lumbar surgery, with pre-operational anticipatory blood loss > 400 mL and without any diseases of blood or endocrinal system were enrolled. They were equally randomized into two groups, the trial group and the control group. Ligustrazine was given to patients in the trial group by intravenous dripping at the dose of 2 mg/kg, 30 min before auto-blood collection, also by mixed in the washing saline and heparinized saline solution to make the final concentration of ligustrazine 0.005%. No ligustrazine was given to patients in the control group. The amount of blood loss and autotransfused were measured and recorded; and patients' venous blood samples for T-lymphocyte subsets (CD3, CD4, CD8) determination and CD4/CD8 ratio calculation were collected at different time points, i. e. before surgery (T0) and at 1 h (T1), 1st day (T2) and 5th day (T3) after ABT.

RESULTSCD3 decreased obviously at T1 and T2 in both groups (P < 0.05 or P < 0.01), while at T3, it restored to baseline in the trial group, but remained at the low level in the control group (P < 0.05). Moreover, levels of CD3 at T1-T3 were lower in the control group than those in the trial group respectively (P < 0.05 or P < 0.01). CD4 decreased obviously at T1 (P < 0.01) in both groups, it recovered at T2 in the trial group, but the recovering in the control group was T3, so comparison of CD4 level between groups showed significant difference at T1 and T2 (P < 0.05 or P < 0.01). As for CD4/CD8 ratio, it decreased obviously at T2 and T3 in the control group (P < 0.05), but unchanged in the trial group, showing statistical difference between groups (P < 0.05).

CONCLUSIONLigustrazine had definite protective effect on T-lymphocyte immunity in patients undergoing ABT, which was presented by the milder inhibition and quicker recovery of immunity.

Adult ; Aged ; Blood Transfusion, Autologous ; Female ; Humans ; Male ; Middle Aged ; Pyrazines ; pharmacology ; T-Lymphocyte Subsets ; drug effects ; immunology ; Young Adult

OBJECTIVETo explore the changes of surface antigen and function of rituximab on dendritic cells derived from patients with Primary immune thrombocytopenia (ITP) to further understand the effective mechanism of immunotherapy.

METHODSThe peripheral blood mononuclear cells (PBMCs) were isolated from remission patients with ITP before and after low-dose rituximab infusion, and the PMNCs were stimulated for 5 days by rhGM-CSF and rhlL-4 in 5% CO2 air at 37°C incubator. Then all of DCs were cultured with TNF-α for 48 hours. The morphology of DCs was monitored under inverted microscope daily, and the surface antigens of the DCs were analysed by flow cytometry, meanwhile the levels of IL-12p70 and TGF-β1 in supernatants were detected by ELISA, mix lymphocyte reaction was performed by MTT assay.

RESULTS(1) Rituximab-treated-DCs showed no obvious tree-like protruding compared with untreated-DCs. The former cells were small and most of nucleus were centric. (2) The expressions of HLA-DR, CD80, CD83 and CD86 on rituximab-treated-DCs \[56.37 ± 3.95)%, (36.41 ± 2.82)%, (30.45 ± 4.61)% and (41.98 ± 4.17)%, respectively\] were significantly lower than those untreated-DCs \[(73.71 ± 7.61)%, (55.14 ± 7.30)%, (80.91 ± 7.09)% and (59.03 ± 3.43)%, respectively\](all P < 0.05), the concentration of IL-12p70 was significantly lower, \[(66.87 ± 4.29)% vs (50.17 ± 14.52)%\], while that of TGF-β1 \[(9.70 ± 0.31)%\] higher than the untreated-DCs \[(2.70 ± 0.36)%\] (P < 0.05). (3) The abilities to activate T cells proliferation of rituximab-treated-DCs reduced compared with untreated-DCs.

CONCLUSIONThe surface antigen of ITP-DCs and the concentration of IL-12p70 reduced after the low-dose rituximab infusion. The abilities to activate T cells proliferation reduced while the concentration of TGF-β1 increased. Rituximab may achieve its therapeutic effect on ITP by downregulating the immunoreactivity of DCs.

Antibodies, Monoclonal, Murine-Derived ; administration & dosage ; therapeutic use ; Cell Proliferation ; Cells, Cultured ; Dendritic Cells ; cytology ; metabolism ; secretion ; Female ; Humans ; Interleukin-12 ; metabolism ; Lymphocyte Activation ; Male ; Rituximab ; T-Lymphocytes ; immunology ; Thrombocytopenia ; drug therapy ; immunology ; metabolism ; Transforming Growth Factor beta1 ; metabolism