Objective:To optimize the processing technology of Moslae Herba processed with ginger juice, and to explore the changes of its volatile components in processing process. Method:The volatile components in Moslae Herba, ginger juice and Moslae Herba processed with ginger juice were extracted by steam distillation. Volatile components in these products were analyzed by HS-GC-MS and identified by NIST 11 standard mass spectra library. Gas chromatographic conditions were as following:HP-5MS elastic quartz capillary column(0.25 mm×30 m, 0.25 μm), helium as the carrier gas, flow rate of 1.0 mL·min^-1, injector temperature at 250℃, sample quantity of 0.2 μL, split ratio of 50:1, temperature program for initial temperature at 40℃, up to 60℃ with the heating rate at 5℃·min^-1, keep 2 min, up to 160℃ with the heating rate at 5℃·min^-1, keep 3 min, finally rise to 250℃ with the heating rate at 25℃·min^-1, keep it for 2 min and finish, mass spectrometry conditions were as following:electron impact ionization(EI), electron collision energy of 70 eV, ion source temperature at 230℃, the interface temperature at 280℃, quadrupole temperature at 150℃, no delay of solvent, electronic multiplier voltage at 2.188 kV, taking full scan mode, scanning range of m/z 35-550.Taking frying time, solid-liquid ratio and moistening time as factors, orthogonal test was adopted to optimize the processing technology with the comprehensive score of relative contents of thymol and carvacrol, number of volatile components and extracting amount of volatile oil as index. Result:A total of 27 volatile components were detected in Moslae Herba. There were 81 volatile components in Zingiberis Rhizoma Recens. The processed products of orthogonal test(No. 1-9) had 31, 38, 29, 35, 38, 33, 34, 22 and 26 volatile components, respectively. Extracting amount of volatile oil was in the order of Moslae Herba processed with ginger juice > Zingiberis Rhizoma Recens > Moslae Herba. The best processing technology was as following:moistening Moslae Herba with equal volume of ginger juice for 6 h, stir-frying for 8 min. Conclusion:Processing has certain impact on the extracting amount of volatile oil in Moslae Herba and the types of volatile components. This optimized technology is stable and feasible, which can provide experimental data for the quality evaluation of processed products of Moslae Herba, and lay a foundation for clarifying its processing mechanism.

OBJECTIVE: To evaluate whether ultrafine particulates (UFPs) have direct deleterious effects on cardiac function through activating MAPK signaling. METHODS: Langendorff-perfused Sprague-Dawley rat hearts were randomly divided into 2 groups (n=10/each group). In control group, the rat hearts were perfused with Tyrode's buffer for 40 min; in UFPs-treated group, the hearts were perfused with UFPs at a concentration of 12.5 mg/L. Cardiac function was determined by measuring left ventricular developed pressure (LVDP), left ventricular peak rate of contraction and relaxation (±dp/dt_max) and coronary flow (CF). The levels of malondialdehyde (MDA), superoxide dismutase (SOD), total anti-oxidant capacity (TAOC) were detected in order to evaluate cardiac oxidative stress via the thiobarbituric acid assay, water soluble tetrazolium salt assay and colorimetry, respectively. The expressions of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium were observed using immunohistochemical staining and Western blots. RESULTS: No significant changes in cardiac function were detected before and after the perfusion in control group while UFPs perfused hearts showed a decline in cardiac function in a time-dependent manner (all P < 0.05). In UFPs-treated group, LVDP, +dp/dt_max, -dp/dt_max and CF were statistically reduced from (82.6±2.1) mmHg, (1 624±113) mmHg/s, (1 565±116) mmHg/s, (12.0±0.2) mL/min to (56.8±4.4) mmHg, (1 066±177) mmHg/s, (1 082±134) mmHg/s, (8.7±0.3) mL/min (all P < 0.05), respectively. Furthermore, The comparison between the two groups observed that UFPs perfusion caused a significant decrease in cardiac function at 30 and 40 min compared with the control group (all P < 0.05). At the end of the perfusion, the level of MDA was increased from (0.98±0.14) nmol/L to (1.95±0.18) nmol/L, while SOD and TAOC were reduced from (12.50±1.87) U/mL and (6.83±1.16) U/mL to (6.50 ±1.04) U/mL and (3.67±0.82) U/mL (all P < 0.001) in UFPs group, respectively. In coincidence with these changes, immunohistochemistry and Western blots results showed that the levels of p-p38 MAPK, p-ERKs and p-JNKs in the myocardium significantly increased in UFPs group as compared with control group (all P < 0.05). CONCLUSION The results of this study demonstrated that the short-term exposure of UFPs to the isolated rat hearts has direct and acute toxic effects on cardiac function, probably related to attenuation of anti-oxidative capacity and activation of MAPK signaling pathways.
Animals ; Heart ; Malondialdehyde/metabolism* ; Myocardium ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the effect of interfering ADAM10 on proliferation and apoptosis of multiple myeloma MM.1S cells, and its possible mechanism.

METHODSFour pairs of shRNA-coding sequences directed against different sites of ADAM10 mRNA were designed and inserted into lentiviral vector plasimd pLVshRNA-EGFP(2A) Puro for constructing the sh/ADAM10-1, sh/ADAM10-2, sh/ADAM10-3, sh/ADAM10-4 and sh/Con. These plasmids and lentiviral packaging plasmids were co-transfected into the packaging cells 293FT, then the virus particles were collected and the viral titer was assayed after concentration, and these viral particles were transfected to MM.1S cells. The flow cytometry was used to sort GFPcells. Real-time quantitative PCR, and Western blot were used to detect the effect of interfering the ADAM10 gene by lentiviral vector mediated shRNA. The proliferation-inhibition curve was plotted by CCK-8 method, the cell viability and apoptosis were detected by flow cytometry with Annexin V and 7-AAD staining, the transcripts of pro-apoptosis gene BAD, BAK, BIK, anti-apoptotic genes BCL-2, c-Myc and Notch1 target gene Hes-1 were detected by real-time PCR.

RESULTSLentivirus vector was successfully constructed, that could specifically interfere ADAM10 expression. Interfering ADAM10 gene could inhibit the MM.1S cell proliferation and induce apoptosis. After the interferencing ADAM10 gene the mRNA levels of pro-apoptosis gene BAD, BAK and BIK were increased, and the mRNA levels of anti-apoptotic genes BCL-2 and c-Myc were reduced. Q-PCR results showed that the mRNA level of Notch1 were increased, but that of Hes-1 were reduced.

CONCLUSIONDown-regulated ADAM10 expression can significantly inhibit multiple myeloma MM.1S cell proliferation and promote the apotosis. Its mechanism may be related to Notch1 signaling pathways.

OBJECTIVE: To evaluate the safety and efficacy of allogeneic natural killer (NK) cells in the treatment of primary hepatocellular carcinoma (HCC), and to elucidate the mechanism of NK cells therapy. METHODS: Twenty-one patients with primary HCC treated with allogeneic NK cells at the Fifth Medical Center of the PLA General Hospital were followed up for 1 year. Peripheral blood mononuclear cells (PBMCs) were isolated from patient-related donors and cultured in vitro for 15 days and infused to the patients in two consecutive days. Clinical data and laboratory data were collected and analyzed, including survival, clinical features, imaging changes, hematology, immunology, and biochemical indicators to evaluate the safety and efficacy of allogeneic NK cell therapy. The changes of peripheral blood lymphocyte subsets after treatment were also analyzed to explore the possible anti-tumor mechanisms. RESULTS: (1) Of the 21 patients with primary HCC, 11 patients were treated once, 5 patients were treated twice, and 5 patients were treated 3 times. After allogeneic NK cells infusion, 10 patients had fever, 1 patient had slight hepatalgia and 1 patient had slight headache, no other adverse events occurred including acute and chronic graft-versus-host disease (GVHD). They resolved spontaneously within 8 hours without other treatment. (2) The total disease control rate was 76.2% during one-year follow-up. Among them, the patients with Barcelona clinic liver cancer (BCLC) stage A had a disease control rate of 100%, stable disease (SD) in 10 cases; BCLC stage B patients had a disease control rate of 60%, partial response (PR) in 1 case, and SD 2 in cases; BCLC stage C patients had a disease control rate of 50%, complete response (CR) in 1 case, and 2 cases of PR. (3) The frequencies of NK cells and CD8+ T cells in peripheral blood were significantly lower than that before at 24 hours after treatment, and the frequencies of CD4+ T cells and CD4/CD8 were significantly higher than the baseline. CONCLUSION Allogeneic NK cells have good safety and efficacy in the treatment of primary HCC. The anti-tumor effect of the allogeneic NK cells may play an important role in the activation of the patient's natural immune system and delay disease progression, suggesting that allogeneic NK cells combined with sorafenib may be a very effective treatment for advanced HCC, and further large-sample multicenter randomized controlled clinical trials are needed to validate this result.
Carcinoma, Hepatocellular ; Graft vs Host Disease ; Humans ; Killer Cells, Natural ; Leukocytes, Mononuclear ; Liver Neoplasms