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 study the effect of omega-3 polyunsaturated fatty acids (omega-3 PUFA) on inflammation in lung tissue of rats with severe scald and its mechanism.

METHODSSeventy-two adult SD rats were divided into sham scald group (SS, n = 8), treatment group 1 (T1, n = 32), treatment group 2 (T2, n = 32) according to the random number table. Rats in T1 group and T2 group were inflicted with 30% TBSA full-thickness scald, and then they were respectively injected with 100 g/L omega-3 PUFA (1 mL/kg) and 200 g/L long-chain fatty acid (2 mL/kg) via tail vein within 5 minutes after burn. The above two fatty acids with equivalent calories were continuously injected for 10 days (once a day). On post burn day (PBD) 1, 4, 7, and 10, serum level of TNF-alpha and level of macrophage inflammatory protein 1alpha (MIP-lalpha) in lung homogenate of T1 and T2 groups were detected, the levels of NF-kappaBp65 and macrophage migration inhibitory factor (MIF) in lung tissue of T1 and T2 groups were observed with immunohistochemical staining (recorded as score). Above-mentioned parameters were also determined in SS group. Data were processed with t test.

RESULTSThe levels of 4 parameters in T1 and T2 groups on PBD 1, 4, 7, 10 were higher than those in SS group (with t values from 3.411 to 8.782, P values all below 0.01), and those in T1 group on PBD 4, 7, 10 were lower than those in T2 group (with t values from 2. 321 to 2.785, P values all below 0.05). The serum level of TNF-alpha and levels of MIP-1alpha, NF-kappaBp65, and MIF in lung tissue in SS group was respectively (0.96 +/- 0.32) ng/mL, (76 +/- 16) pg/mL, 0.24 +/- 0.03, 1.31 +/- 0.03, and those in T1 and T2 groups all peaked on PBD 7 [(2.43 +/- 0.32) ng/mL, (210 +/- 56) pg/mL, 4.23 +/- 2.15, 4.69 +/- 1.83; (3.15 +/- 0.54) ng/mL, (274 +/- 64) pg/mL, 5.15 +/- 2.31, 5.37 +/- 2.16].

CONCLUSIONSOmega-3 PUFA can effectively reduce serum level of TNF-alpha and levels of MIP-1alpha, NF-kappaBp65, and MIF in lung tissue of rats with severe scald, showing that it has a protective effect against injury of lung tissue.

Animals ; Burns ; metabolism ; pathology ; therapy ; Chemokine CCL3 ; metabolism ; Fatty Acids, Omega-3 ; therapeutic use ; Female ; Inflammation ; metabolism ; Intramolecular Oxidoreductases ; metabolism ; Lung ; metabolism ; Macrophage Migration-Inhibitory Factors ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; blood

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

ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Cytotoxicity, Immunologic ; Dendritic Cells ; physiology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; Immunophenotyping ; K562 Cells ; Leukemia ; drug therapy ; immunology ; Lymphocyte Culture Test, Mixed

OBJECTIVETo explore the expression and sequence of ERCC1 gene in CdCl2-induced transformed human bronchial epithelial 16HBE cells at different stages.

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemcial staining (SP method) were used to measure the ERCC1 mRNA and protein expression in 16HBE cells at different passages treated with CdCl2 (the 5th, 15th, 35th passage, and neoplastic cells from tumors formed in nude mice). ERCC1 exon 3,exon 4 of the 16HBE cells and tumor cells from nude mice were amplified by polymerase chain reactions (PCR), the amplified DNA strips were purified,and the exons were detected by DNA analysis.

RESULTSDuring the passages of 16HBE cells treated with CdCl2, the expression of ERCC1 gene was decreased gradually. The ERCC1 gene mRNA and protein expression levels of the CdCl2-transformed 35th passage 16HBE cells and tumor cells from nude mice were significantly decreased comparing with those in non-transformed 16HBE cells (P < 0.01). In the CdCl2-induced tumorigenic cells in nude mice, there was adenine (A) deletion in 1st site of ERCC1 exon 4. The mutation was frame shift mutation.

CONCLUSIONThe decreased expression and mutation of ERCC1 gene may be the possible carcinogenic mechanism of CdCl2.

Animals ; Bronchi ; cytology ; Cadmium Chloride ; toxicity ; Cell Transformation, Neoplastic ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; metabolism ; Endonucleases ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Exons ; Frameshift Mutation ; Humans ; Mice ; Mice, Nude ; RNA, Messenger ; metabolism