OBJECTIVE:To establish a rapid determination method for 9 kinds of glucocorticoid and 8 kinds of non-steroidal anti-inflammatory drugs illegally added into anti-rheumatic TCM patches. METHODS:UPLC-MS/MS method was adopted. The samples were separated on Acquity UPLC? BEN C18 column with methanol-water(90:10,V/V)as mobile phase at flow rate 0.4 mL/min. The column temperature was set at 30℃,and sample size was 1μL. The ionization mode was electrospray ion source(ESI):spray needle voltage of 5500 V,ion source spray GS1,GS2 pressure of 50,50 psi,ion source temperature of 500 ℃,scanning range of m/z 50-500. The multiple reaction monitoring(MRM)combined with information-dependent acquisition (IDA) and en-hanced product ion scanning(EPI)was used to rapid qualitative analysis. RESULTS:The detection limits for 17 kinds of chemical components were not more than 0.400 μg/mL. RSDs for precision,repeatability,stability tests were not more than 4.93%,7.69%, 9.57%,respectively. Diclofenac sodium was detected in the sample. CONCLUSIONS:The method is selective and sensitive. It is suitable for qualitative analysis of chemicals illegally added into anti-rheumatism TCM patches.

Objective:To summarize the complications' prevention and nursing experience for patients after autologous transplantation of the microvascular submandibular gland for severe keratoconjunctivis sicca.Methods:Retrospective analysis of 141 patients(151 sides) after autologous microvascular submandibular gland transfer operation was undertaken to record the timing and incidence of major complications and to summarize the nursing experience.Results: The main complications after the transplantation were: vascular crisis,catheter obstruction,salivary fistula and temporary paralysis of hypoglossal nerve.Related nursing measures in response to these complications were discussed.Conclusion:Targeted nur-sing interventions for patients with autologous transplantation the submandibular gland have a positive effect on the prevention of the surgical complications.

Objective To evaluate the role and efficacy of preventing bone mineral loss in patients with endometriosis treated by gonadotrophin-releasing hormone analogues (GnRH-a) combined with addback therapy.Methods Prospective,randomized controlled studies of the use of GnRHa with add-back therapy in treatment of endometriosis were enrolled in this study from Medline,Embase,Cochrane library,China National Knowledge Internet (CNKI),Chinese Biological Medicine Disk (CBM) and Data Base of Wanfang.After quality assessment and data extraction,meta-analysis were conducted in the change of BMD,reproductive hormone (E2) and visual pain score (VAS) by Stata 11.0 software.Results A total of 785patients from 13 randomized controlled trail (RCT) studies enrolled in this study after exclude no following up,poor quality and repeat published studies.377 patients were in group of GnRH-a with add-back treatment and 408 patients were in group of GnRna alone.The findinds were showed in meta-analysis:(1) there was a significant difference in percentage change of bone mineral density (BMD) between two groups,the addback therapy was more effective in prevention of bone loss which was (SMD =0.223,95％ CI:0.003 to 0.443,P =0.047).(2) There was no significant difference in the level of reproductive hormone between two groups (SMD =-0.053,95％ CI:-0.479 to 0.373,P =0.807).(3) There was also no significant difference in the visual pain score between the two groups (SMD =-0.157,95％ CI:-0.474 to 0.160,P=0.332).Conclusions GnRH-a with add-back therapy have been shown to be more effective in preventing loss of BMD than GnRH-a treatment alone.However,the long term effect of preventing BMD should be studied.

Objective To evaluate the correlation between intracranial hyperintense vessel sign (HVS) on fluid-attenuated inversion recovery(FLAIR) and the degree of the stenosis of internal carotid artery (ICA).The effect of carotid endarterectomy(CEA) on HVS was assessed.Methods A retrospective analysis of MR FLAIR sequence and cerebral-cervical computed tomography angiography(CTA) was performed in 1 total of 491 patients.Of the 491 patients,41 treated with CEA were evaluated using their pre-and post-operative image data.Patients were divided into ICA stenosis group and non-stenosis group according to the CTA imaging findings.The ICA stenosis group was subdivided into unilateral group and bilateral stenosis group.Furthermore,we measured and graded the ICA of the unilateral stenosis group into seven stenotic degrees,they were＜50％(n=40),50％-＜60％(n=15),60％-＜70％(n=17),70％-＜80％ (n=6),80％-＜90％ (n=7),90％-＜100％ (n=23),100％ (n=24),respectively.Chi square test was used to analyze the occurrence rates of HVS between ICA stenosis and non-stenosis group,and between ICA unilateral and bilateral stenosis group,respectively.Spearman rank correlation was performed to evaluate the correlation between the presence of HVS and stenotic degrees of the ICA.For the 41 patients who underwent CEA,pre-and post-operative image data were compared,focusing on the presence or disappearance of the HVS on MR FLAIR imaging.Results HVS on FLAIR images were observed in 81 of 177 patients(45.76％) with ICA stenosis,and in 59 of 314 patients(18.79％) without ICA stenosis.The occurrence rate of HVS was significantly higher in patients with ICA stenosis than those without ICA stenosis (x2=40.40,P＜0.01).There was no statistical significance in the occurrence rates of HVS between ICA unilateral stenosis group and bilateral stenosis group(x2=0.24,P＞0.05).The occurrence rates of HVS of ICA graded as the seven stenotic degrees were 22.00％(8/40),26.67％(4/15),35.29％(6/17),33.33％ (2/6),42.86％ (3/7),69.57％ (16/23),83.33％ (20/24),respectively.There was a significant positive correlation between the occurrence rates of HVS and the degrees of ICA stenosis(r=0.964,P＜0.01).HVS disappeared in 19(86.36％) out of 22 patients with HVS on pre-operative MR images after CEA in The remaining HVS in 3(13.64％) patients was attributed to the failure of completely recanalization of ICA.Conclusions A close relationship exists between HVS and ICA stenosis.The presence of HVS indicates a high probability of the existence of severe ICA stenosis.A further assessment for ICA stenosis is warranted.HVS disappearance after successful CEA indicates that HVS can be a useful marker for the the evaluation of consequence associated with CEA.

Histiocytic Necrotizing Lymphadenitis ; Humans

Objective To explore the dose-dependent and time-dependent effect of docetaxel on the expression of mammalian eukaryotic initiation factor 3 subunit A (eIF3a) in lung cancer cell line. Methods The human lung cancer cell line A549 was treated with gradient concentrations of docetaxel for different time. Real-time PCR and Western blot were used to detect mRNA and protein expression levels of eIF3a and α-tubulin, respectively. Results Docetaxel did not affect α-tubulin expression at either mRNA level or protein level. When A549 cells were treated with high concentration of docetaxel (30 μg/L), the expression level of eIF3a mRNA tended to increase in a time-dependent manner. Protein expression level of α-tubulin was not associated with eIF3a expression significantly in cells treated by docetaxel.Conclusion Docetaxel could slightly increase the expression of eIF3a mRNA, and eIF3a does not regulate the expression of α-tubulin in A549 cells treated by docetaxel.

BACKGROUND:Lymph-targeted tracing and therapy based on carbon nanotubes have been one of the hottest researches on targeting tumor diagnosis and treatment. To evaluate the accumulation of carbon nanotubes in axil ary lymph node can provide experimental evidences for developing nano-tracers and drug carriers which are more lymph-specific and more biocompatible. OBJECTIVE:To study the accumulation of the intravenously injected carboxylated single-wal ed carbon nanotubes in axil ary lymph nodes of Sprague-Dawley rats, and to evaluate their effect on blood cel s. METHODS:Sixty-four Sprague-Dawley rats were randomly divided into two groups. Rats in testing group were injected with carboxylated single-wal ed carbon nanotubes suspension (2 mg/kg), while those in control group were injected with 5%glucose solution (1 mL/kg), both through the tail vein, three times per week. Four periods of 7, 60, 90 and 120 days were set (the 120-day period referred to 90 days of administration fol owed by 30 days of drug withdrawal). At the end of each period, eight rats from each group were randomly picked out, to col ect blood samples via the abdominal aorta for blood routine test. Final y the axil ary lymph nodes were observed, and the lymph node samples of rats in the testing group were col ected and analyzed at 120 days by transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, black staining of axil ary lymph nodes of rats in testing group was not obvious at the end of the 7-day period. However, with the increase of the dosing periods, the lymph nodes of the rats in the testing group became enlarged, firm and black stained, coupled with a significant rising in the percentage of blood neutrophils. After 30 days of drug withdrawal, the size of the rat axil ary lymph nodes was reduced and black staining partly faded, with the decreasing of blood neutrophil percentage. Under the transmission electron microscope, abundant carboxylated single-wal ed carbon nanotubes were uptaken by lymphocytes to form a large number of phagocytic vacuoles after drug withdrawal for 30 days. It indicates that the short-term tracing of rat axil ary lymph nodes by carboxylated single-wal ed carbon nanotubes injected through the tail vein is relatively weak, while the long-term intravenous injection can cause their accumulation in rat axil ary lymph nodes, coupled with the increase of neutrophils;after drug withdrawal, the carboxylated single-wal ed carbon nanotubes can be slowly cleared by the lymph nodes.

OBJECTIVE:To improve the dispersion stability in water of Tussilago farfara powder,and to improve compliance of Xiao’er feike granules. METHODS:The effects of 4 kinds of dispersion stabilizer (sodium hexametaphosphate, dextrin, PEG4000 and lecithin) on dispersion stability of suspension in water were investigated during the grinding of T. farfara using rate of absorbance change(β)and Zeta potential as index;IR spectrum of samples were characterized. Using original formulation with-out dispersion stabilizer as control,the dispersion stability of new formulation granules in water were analyzed comparatively after adding dispersion stabilizer. RESULTS:Among 4 kinds of dispersion stabilizer,β of sample prepared by sodium hexametaphos-phate was the lowest,while Zeta potential of it was the highest;compared with original T. farfara,β of T. farfara grinded with 2.5% sodium hexametaphosphate decreased by 16.8%,and Zeta potential absolute value increased by 29.4%;no new peak was found in IR spectrum. Compared with control granules,granules suspension prepared by new formulation had lower β and higher Zeta potential absolute value (P<0.01);particle size was 30 μm and no large particle aggregation was found;β was less than 5.0% within 20 s sedimentation. CONCLUSIONS:During the preparation of Xiao’er feike granules,the application of sodium hexametaphosphate in the grinding of T. farfara powder can improve the dispersion stability of granules in water and the compliance of the preparation.

Aim To investigate the role of GC in indu-cing apoptosis of amygdaloid neurons.Methods Cul-turing primary neurons of amygdala,the neurons were identified by immunefluorescence techniques with anti-body against microtubule associated protein-2 (MAP2 ) and antibody against GC receptor.Using flow cytome-try to detect the effects of different concentrations of dexamethasone on the amygdala neuron apoptosis. Then the experiment was divided into four groups:CON ,DEX ,DEX +MIF and MIF .The rate of apopto-sis of the four groups was detected by TUNEL tech-nique and the expressions of BAX mRNA of four groups by Real-time PCR technique.Results (1 )Compared with the control group, the percentage of apoptotic cells increased significantly with DEX(10 -8 mol·L-1~10 -6 mol · L-1 )treatment in a concentration-de-pendent manner.(2)the TUNEL test showed that the percentage of apoptotic cells of DEX group increased significantly,compared with control group.While it decreased significantly in DEX+MIF group,compared with DEX group.There was no difference between MIF group and control group.(3 )Compared with control group,the expressions of BAX mRNA of DEX group increased significantly.While the expressions of BAX mRNA of DEX +MIF group decreased significantly, compared with the DEX group.There was no difference between MIF group and control group.Conclusion GC can independently induce the apoptosis of primary cultured neurons in the amygdala by combining with GC receptor.

Objective To investigate the effect of Euphorbia helioscopia on MDA-MB-231 cells and its mechanism.Methods The cell viability was detected by MTT assay.The production of ROS in MDA-MB-231 cells was measured by fluorescence microscopy.The apoptotic rate was detected by flow cytometry.Apoptosis DNA fragments were detected by TUNEL assay.Western blot was used to assess the expression of caspase-9, caspase-3 and PARP.Results MTT assay showed that the extract significantly inhibited the viability of MDA-MB-231 cells, which can be diminished by the ROS inhibitor NAC and the caspase inhibitor Z-VAD-FMK.The marked increase in the production of ROS induced by the extract was observed with fluorescence microscopy.Flow cytometry showed that the PI positive staining cells increased significantly after the treatment of the extract, but was diminished by NAC.Caspase-9 and caspase-3 were activated after the treatment of the extract while the PARP was cleaved.TUNEL showed that a significant increase in apoptotic DNA fragmentation induced by the extract, which can be diminished by NAC and Z-VAD-FMK.Conclusion Ethyl acetate extract inhibited the MDA-MB-231 cells and induced apoptosis.The mechanism may involve with the mitochondrial damage due to the excessive ROS.