This study aimed to explore Semaphrin4D (Sema4D) expression and clinical significance in non-small cell lung cancer (NSCLC), and to define the roles and mechanisms of Sema4D in regulating the malignant behaviors of A549 cells by small interfering RNA (siRNA). Firstly, immunohistochemistry revealed that Sema4D was more frequently expressed in NSCLC than in lung benign lesion (P<0.05) and its overexprssion was associated with low differentiation (P<0.05), poor pTNM staging (P<0.05) and occurrence of lymph node (LN) metastasis (P<0.05). Endogenous Sema4D expression was suppressed by Sema4D siRNA in A549 cells overexpressing Sema4D. Protein levels of Sema4D, total Akt and p-Akt were examined by Western blotting. Cell proliferation, migration and invasion abilities were measured by MTT assay and Transwell assay respectively. Results showed that Sema4D siRNA significantly suppressed phosphorylation of AKT in A549 cells, but it did not alter total AKT expression. In addition, efficient down-regulation of SemaD significantly inhibit cell proliferation (P<0.05), migration (P<0.05) and invasion (P<0.05) in A549 cells. These findings suggest that Sema4D might serve as a reliable tool for early prediction of NSCLC poor prognosis. Sema4D could play an important role in promoting tumor proliferation, migration and metastasis in the NSCLC, by influencing the Akt protein phosphorylation. Inhibition of Sema4D may be a useful approach for the treatment of NSCLC.

BACKGROUND:There is little evidence on the use of al ogeneic tendon graft in the reconstruction of ankle joint.
OBJECTIVE:To explore the clinical outcome of anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon graft in patients with chronic ankle instability.
METHODS:Twenty-six patients with chronic lateral instability underwent anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon. There were 18 cases of simultaneous injury or chalasia in calcaneofibular ligament and anterior talofibular ligament, and 8 cases of anterior talofibular ligament injury or chalasia. The ankle joint function was evaluated according to AOFAS scale and Good classification. The affect ankle and healthy ankle were compared in the extension, plantar flexion activity, and metaleg activity.
RESULTS AND CONCLUSION:Al the 26 patients were fol owed up for 9-24 months with a mean of 15 months. No cases appeared recurrent ankle lateral instability. The mean AOFAS score in the group of calcaneofibular ligament and anterior talofibular ligament was improved from (48.4±3.7) points preoperatively to (88.2±3.8) postoperatively, while that in the group of anterior talofibular ligament was improved from (50.0±6.4) points preoperatively to (89.5±3.4) points postoperatively. According to Good score, there were excellent in 19 feet, good in 6 feet, fair in 1 foot, with an excellent and good rate up to 96%. No serious complication was occurred in this group. Anatomical reconstruction of the lateral ligaments with cryopreserved al ogeneic tendon graft can increase the tendon-bone contact area, improve the rate of tendon healing, and enhance the stability of ankle joint in patients with chronic ankle instability. Further studies are needed to verify its long-term efficacy.

Objective To develop a HPLC-MS/MS method for the determination of aconitine and study thein vitro metabolic stability of aconitine in dog tissue homogenates.Methods The chromatographic separation was performed on a C18 column.The mobile phase consisted of acetonitrile and water with 0.2％ formic acid and 5 mmol/L ammonium acetate.A triple quadrupole tandem mass spectrometer equipped with an electrospray ionization interface source was used for the quantitative determination in the positive selective reaction monitor mode.Aconitine was incubated with dog tissue homogenates and samples were withdrawn at different time points and precipitated by acetonitrile with internal standards citalopram.Results Aconitine showed good linear relationship over the range from 5 to 500 ng/ml.The recoveries of aconitine were between 85.73％ and 92.12％ at three QC concentration levels.The intra- and inter-day precisions were 5.32％ - 8.95％ and 5.45％ - 8.86％,respectively.After incubation,about 20％ of aconitine were cleared in the liver and small intestine,and t1/2 were 460.6 and 521.3 min,respectively.But none was metabolized in the stomach and kidney.Conclusion These results demonstrated that aconitine was mainly metabolized in the liver and small intestine at a slow rate.

An analytical method was developed for the simultaneous extraction and determination of eighteen fluoroquinolones (FQs), tetracyclines (TCs) and sulfonamides (SAs) antibiotics from soils using solid phaseextraction and liquid chromatography tandem mass spectrometry.Soil sample was firstly extracted by phosphate buffer at pH 3 in combination with 50% of organic modifier acetonitrile, then purified and concentrated by SAX and HLB column.Qualitative and quantitative analysis were carried out for the analyte under the MRM mode after the chromatography separation on Kromasil C_(18)(250 mm x4.6 mm, 5 μm) column.The range of recoveries (in percent) for FQs, TCs, SAs, in the soil matrix was 67.20%-88.98%, 62.23%-85.36%, 55.76%-97.37% with 1.1%-17.2% of relative standard deviation respectively in two different concentra tions.The limits of quantification (LOQ, S/N = 3) were 3.36-8.88 jig/kg, 0.56-0.91 μg/kg and 0.07-1.85 μg/kg for FQs, TCs and SAs, respectively.This method was successfully used to detect 18 anti biotics in 6 soil samples with different land types in Tianjin.Results showed some of the antibiotics in the arable soil were detected, with concentrations of 1.72-119.57 μg/kg.

Objective To evaluate the effects of the chronic Keshan disease's self-management in Liangshan Prefecture of Sichuan Province. Methods According to "The Serf Management Program of Chronic Keshan Disease in Liangshen Prefecture of Sichuan Province", 56 chronic Keshan disease patients were selected in the personalized self-manngemant evaluation under the instructions by endemic disease specialists and the rural doctors. Evaluation was based on changes of indexes such as the clinical symptoms, general health conditions, the electrocardiogram, X-ray, the heart function, etc, before treatment and 3 and 6 months following the treatment. Results Clinical signs and symptoms of the patients were significantly improved 3 and 6 months after the treatment, and the improvement was more obvious 6 months than 3 months following the treatment(P<0.05 or< 0.01). After treatment for 3 months, the patients'electrocardingram and heart function did not show obvious change (X2=0.05,039, P0.05); hut obvious improvements Eexcept X ray results(X2=0.61 ,P0.05)] were seen 6 months after treatment (X2=4.36,16.84, P<0.05 or<0.01). Altogether, among the 56 patients evaluated after treatment for 6 months, none achieved the clinical cure standard, 26 cases(46.3%) showed significant improvement, 17 cases (30.4%) were stable, 5 cases (8.9%) were aggravated,one case (1.8%) lost contact, and 6 cases (10.6%) died. Conclusion The project of the chronic Keshan disease's self-management is suitable for the present situati,on of the endemic regions and can he introduced to many places in our country.

OBJECTIVETo observe the effects of Shenfu Injection (SI) on immune function in patients during perioperative cardiopulmonary bypass (CPB).

METHODSNinety patients scheduled for CPB were randomly divided into 3 groups, the group A treated with SI before CPB, the group B treated with SI after CPB and the group C (the control group), 30 cases in each group. Counts of leukocyte, neutrophil, lymphocyte and T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) and serum levels of IgG, IgA, IgM, C3 and C4 were investigated at different time-points, i.e. one day before CPB, immediately after anesthesia and finishing CPB, and the 1st, 3rd and 8th post-operational days.

RESULTSIn all the three groups, counts of leukocyte and neutrophil rose immediately after CPB and the high level maintained on the 1st and 3rd post operational days, it somewhat lowered on the 8th day but still higher than that before operation, showing insignificant difference among the 3 groups. As for the other indexes, they declined obviously and immediately after CPB and began to restore on the 8th day to a certain extent, and the restoration were better in the group A and B, as the changes were milder.

CONCLUSIONSI could improve the immune function and reduce the risk of post operational infection in patients during perioperative CPB.

Adjuvants, Immunologic ; therapeutic use ; Adult ; Aged ; Cardiopulmonary Bypass ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Heart Diseases ; immunology ; surgery ; Humans ; Injections, Intravenous ; Intraoperative Period ; Male ; Middle Aged ; Phytotherapy ; Postoperative Complications ; prevention & control ; T-Lymphocyte Subsets ; drug effects

Objective To analyze characteristics of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype B/C infectors. Methods 19 HIV-1 recombinant subtype B/C infectors infected within 1 year, 40 chronic infectors infected for more than 3 years were enrolled in this cohort study. Elispot assay was used to observe HIV-1 specific T lymphocyte responses in HIV-1 recombinant subtype B/C infectors. Results Nef-specific T lymphocyte responses of interferon-gamma secretion were identified in 15 Chinese HIV-1 recombinant subtype B/C infectors infected within 1 year. The specific T lymphocytes were mainly targeted at four peptides which span from Nef 83 to 135: EVA7081.1, EVAT081.5, EVA7081.6 and EVAT081.48. Responses were identified in 29(75. 2%) infectors with more than 3 years of infection and the specific T lymphocytes were mainly targeted at three peptides which span from Nef 63 to 101 : EVA7081.43, EVA7081.44, EVAT081.45, EVA7081.47, EVA7081.48 and EVA7081.49. The average magnitude of response in infectors with less than 1 year of infection was 284. 13 SFC/106 PBMC. The average magnitude of response in infectors with more than 3 years of infection was 152. 44 SFC/106 PBMC. There was a significant difference between the two groups (U = 91. 000, P = 0. 002). Conclusions HIV-1recombinant subtype B/C infectors at different stages of diseases (less than 1 year and more thank 3 years) can recognize central region of Nef. The magnitude of Nef-specific IFN-γ secretion T lymphocyte responses in this cohort gradually decrease with disease progression.

Objective To analyze character of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype C/B' and subtype B' infectors and to identify the common immunodominant re-gions recognized by these infectors. Methods Fifty-nine HIV-1 recombinant subtype C/B' infectors and 27 subtype B' infectors were tested by IFN-γ enzyme linked immunospot (ELISPOT) assay using HIV-1 C/B' Nef overlapping peptides. Results Nef-specific T-cell responses of IFN-γ secretion were identified in 44 (74.58%) out of 59 HIV-1 recombinant subtype C/B' infectors. Ten peptides, EVA7081.1,5, 6, 7,43, 44, 45, 47, 48, 49 were mainly recognized. Amino acid position was from Nef63 to 115 and 117 to 139. Twenty of 27 (74.07%) HIV-1 subtype B' infectors recognized peptides. EVA7081.1,2, 43 and 49 were mainly recognized. Amino acid position was from Nef 63 to 77 and 87 to 119. There was no correlation be-tween the Nef-specific IFN-production of HIV-1-specific T cells responses and viral load or CD4 T cell count in both subtype infectors. Conclusion The immunodominant regions, from Nef63 to 77 and 87 to 115 were recognized by both Chinese HIV-1 recombinant subtype C/B' infectors and subtype B' infectors. These re-gions could be used in design of vaccine.

This study aimed to explore Semaphrin4D (Sema4D) expression and clinical significance in non-small cell lung cancer (NSCLC), and to define the roles and mechanisms of Sema4D in regulating the malignant behaviors of A549 cells by small interfering RNA (siRNA). Firstly, immunohistochemistry revealed that Sema4D was more frequently expressed in NSCLC than in lung benign lesion (P<0.05) and its overexpression was associated with low differentiation (P<0.05), poor pTNM staging (P<0.05) and occurrence of lymph node (LN) metastasis (P<0.05). Endogenous Sema4D expression was suppressed by Sema4D siRNA in A549 cells overexpressing Sema4D. Protein levels of Sema4D, total Akt and p-Akt were examined by Western blotting. Cell proliferation, migration and invasion abilities were measured by MTT assay and Transwell assay respectively. Results showed that Sema4D siRNA significantly suppressed phosphorylation of AKT in A549 cells, but it did not alter total AKT expression. In addition, efficient down-regulation of SemaD significantly inhibit cell proliferation (P<0.05), migration (P<0.05) and invasion (P<0.05) in A549 cells. These findings suggest that Sema4D might serve as a reliable tool for early prediction of NSCLC poor prognosis. Sema4D could play an important role in promoting tumor proliferation, migration and metastasis in the NSCLC, by influencing the Akt protein phosphorylation. Inhibition of Sema4D may be a useful approach for the treatment of NSCLC.
Aged ; Antigens, CD ; biosynthesis ; Carcinoma, Non-Small-Cell Lung ; metabolism ; mortality ; pathology ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Disease-Free Survival ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; mortality ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Semaphorins ; biosynthesis ; Survival Rate

Amino Acid Sequence ; Base Sequence ; Klebsiella pneumoniae ; drug effects ; enzymology ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; beta-Lactamases ; chemistry ; genetics