AIM: To investigate the effects of Qiwei Qianliening Tablets(QLN)(Radix paeoniae rubra,Semen Vaccariae,Semen Citri reticulatae,etc) on nonbacterial chronic prostatitis and its anti-inflammatory and analgesia activity. METHODS: Nonbacterial chronic prostatitis model was induced by injecting 25% Xiao Zhiling Injection into the prostate of rat and then coefficent of prostate and pathologic changes were observed.Anti-inflammatory effects were tested by cotton pelle-induced granuloma in rat.Analgesia activity was tested by hot-plate method. RESULTS: QLN inhibited the increase of prostate tissue weight and the histopathology showed resolving prostatitis in QLN-treated groups compared with the controls.Anti-inflammatory activity was confirmed in cotton pelle-induced granoluma in rat and QLN-treated groups extended the licking-paw latency in hot-plate method compared with the controls. CONCLUSION: Qiwei Qianliening Tablets has prevention and treatment effect on experimental nonbacterial chronic prostatitis.

AIM:To observe the influence of high expression of miR-15a-5p on the proliferation and migration of human hepatocellular carcinoma SMMC-7721 cells.METHODS: The miR-15a-5p oligonucleotide , which was recon-structed with additional restriction sites of EcoRⅠand HindⅢ, was chemically synthesized and confirmed by sequencing . The miR-15a-5p eukaryotic expression system was constructed by pcDNA 6.2-GW/Em-GFP-pre-miR-15a-5p plasmid.The miR-15a-5p was transfected into the SMMC-7721 cells transiently by plasmid , and quantified by quantitative real-time PCR at the mRNA level.The cell viability was measured by CCK-8 assay, and the living cell counting was performed by the method of Trypan blue exclusion .The migration ability of the SMMC-7721 cells with high expression of miR-15a-5p was de-tected by wound healing test .RESULTS: The sequence of miR-15a-5p oligonucleotide 100% matched the designed se-quence.Compared with control group , the miR-15a-5p expression was increased significantly (P<0.05).The viability, the living cell number and the migration ability of the SMMC-7721 cells were decreased in high expression of miR-15a-5p group with statistically significant difference (P<0.05).CONCLUSION: The abilities of proliferation and migration in human hepatocellular carcinoma SMMC-7721 cells are decreased by high expression of miR-15a-5p.

Objective To establish plaque reduction assay and evaluate the activities of oseltamivir (tamiflu),amantadine,ribavirin and herb radix isatidis against influenza virus in vitro.Methods Plaque reduction assay was used to determine IC_(50) values of four studied drugs above in this susceptibility testing in which 8 clinical isolates(three influenza A virus isolates and five influenza B virus isolateds)were inoculated and tested.Results By testing of 8 clinical isolates of influenza virus A and B isolated between the year 2001 to 2008,oseltamivir and amantadine were found to be sensitive to influenza A virus with IC_(50) of 0.064 -0.128 mg/L and 0.5 mg/L,respectively.However,ribavirin(IC_(50)>8 mg/L)was not found to be sensitive,and herb radix isatidis had totally no activities.Unfortunately.all four studied drugs were not found to have activities against influenza B virus in vitro.Conclusions It Was indicated that oseltamivir and amantadine.but not ribavirin and herb radix isatidis.are sensitive to influenza A virus.All four studied drugs were not found to have activities against influenza B virus in vitro.

Objective Experimental model of experimental autoimmune Myasthenia Gravis (EAMG) were established to explore the effect of Zini-Huangqi-Gegen decoction and Peiyuan-Guben powder on EAMG rats model.Methods Experimental animals were randomly divided into the control group (n=10) and the model group (n=30).The model rats were induced by murine AChR-α97-116 peptide immunostaining for EAMG rats model.After the first immunization,the general condictions and body weight of rats were observed,and the Lennon score was used to evaluate the rats.The second immunization was performed on the 1 1th day after the first immunization.On the 15th day after the first immunization,the rats were randomly divided into the model group(n=8),Huangqi-Gegen decoction and Peiyuan-Guben powder group (abbreviated Huangpei group,n=8) and Prednisone group (n=8) according to the Lennon score.Huangpi group rats were treated with Huangqi-Gegen decoction (21.5 g/kg) combined with Peiyuan-Guben powder (0.8 g/kg),prednisone group with 0.005 4 g/kg prednisone aqueous solution,the control group and the model group oral volume of distilled water.The rats were administered with a body weight of 10 ml/kg once a day for a total of 56 days.At the 70th day after the first immunization,serum was extracted from the rats.The Anti-AChR-α97-116 IgG and its subtype in serum were detected by ELISA.The IL-4,IL-10,IL-17 in serum were detected by ELISA.Results Compared with the model group,the weight of the rats in the Huangpei group and the prednisone group significantly increased after the 28th day of the first immunization (P＜0.05).After the 36nd day of the first immunization,the Lennon score of the Huangpei group significantly decreased (P＜0.05).At the end of the administration,the amplitude of EMG amplitude attenuation (41.83％ ± 7.45％ vs.67.76％ ± 4.32％) in the Huangpei group significantly decreased (P＜0.05),and the serum IgG (1.15 ± 0.07 vs.1.24 ± 0.08),IgG1 (0.17 ± 0.01 vs.0.25 ± 0.03),IL-4 (16.54 ± 1.66 pg/ml vs.25.64 ± 1.74 pg/ml),IL-10 (113.65 ± 12.87 pg/ml vs.121.54 ± 10.44 pg/ml),IL-17 (43.58 ± 3.54 pg/ml vs.65.76 ± 3.59 pg/ml) in the rat serum significantly decreased (P＜0.05).Conclusions Huangqi-Gegen decoction and Peiyuan-Guben powder can increase the body weight of rats,decrease the concentration of AChR-Ab in serum,the concentrations of IL-4,IL-10 and IL-17 in serum,and effectively improve the symptoms of EAMG rats.

Objective To investigate the effect of Yanghe decoction serum on the proliferation of breast cancer stem cells HMLER90hi and its mechanism. Methods Twenty female Sprague-Dawley rats were randomly divided into low,medium and high dose groups containing drug serum and control group, in order to prepare the Yanghe decoction serum and blank serum. After 24 hours of drug intervention,the effects of each group on the proliferation of HMLER90hi cells at 24 h,48 h,and 72 h were detected by MTT assay. The expression of EphA4 and p50 mRNA in each group were detected by real-time fluorescence quantitative PCR. Results Compared with the blank control group,the cell proliferation activity of each dose group of Yanghe decoction significantly decreased at 24 h (0.818 ± 0.061, 0.706 ± 0.073, 0.587 ± 0.052 vs. 0.928 ± 0.075), 48h (0.760 ± 0.047, 0.638 ± 0.056, 0.510 ± 0.059 vs. 0.973 ± 0.095), and 72 h (0.672 ± 0.102, 0.508 ± 0.092, 0.448 ± 0.048vs.1.023 ± 0.099) (P<0.05 orP<0.01), respectively. After 24 h of drug intervention, compared with the control group, the expression of EphA4 mRNA (0.54 ± 0.07, 0.54 ± 0.07, 0.33 ± 0.04 vs.0.68 ± 0.09) significantly decreased, and p50 mRNA (0.69 ± 0.10, 0.54 ± 0.08, 0.41 ± 0.06vs. 0.85 ± 0.13) significantly decreased in each dose group of Yanghe decoction (P<0.05 orP<0.01).ConclusionsTheYanghe decoction can inhibit the proliferation of breast cancer stem cell HMLER90hi,and its mechanism may be related to its inhibition of the conduction of the juxtacrine pathway of monocyte macrophage.

ObjectiveTo investigate the effects of Yanghetang （YHT） on breast cancer 4T1 cells and their mitogen-activated protein kinase （MEK）/extracellular signal-regulated kinase （ERK） signaling pathway. MethodThe YHT-containing serum was prepared from SD rats. The rats were randomly assigned into a blank group （normal saline） and low-， medium-， and high-dose （5.8， 11.6， 23.2 g·kg^-1， respectively） YHT groups. The serum containing 10% YHT in each group was mixed with 90% RMPI 1640 complete medium， and the mixture was used to interfere with the cells. Cell counting kit-8 （CCK-8） method was used to detect the proliferation of the 4T1 cells treated with YHT for 24， 48， 72 h. The apoptosis， migration， and invasion of 4T1 cells were detected by flow cytometry， scratch test， and Transwell assay， respectively. Western blot was employed to determine the expression levels of MEK1/2， phosphorylation （p）-MEK1/2， ERK1/2， p-ERK1/2， and rat sarcoma virus （RAS） protein. ResultCompared with the blank group， the intervention with YHT-containing serum for 24， 48， and 72 h had significant inhibitory effect on 4T1 cell proliferation （P<0.05， P<0.01）. After intervention with YHT-containing serum for 48 h， the apoptosis rate of cells increased （P<0.01）. Compared with the blank group， the intervention with YHT for 24 h and 48 h decreased the healing ability of cells in the scratch test （P<0.01）. The invasive ability of cells treated with the low， medium， and high-dose YHT containing serum showed a decreasing trend （P<0.01）. Compared with the blank group， YHT-containing serum did not change the expression of MEK1/2 and ERK1/2 while down-regulating the expression of p-MEK1/2， p-ERK1/2， and RAS protein （P<0.01）. ConclusionYHT can inhibit the proliferation， migration， and invasion and promote the apoptosis of breast cancer 4T1 cells. In may promote the apoptosis by inhibiting the MEK/ERK signaling pathway and down-regulating the expression of p-MEK1/2， p-ERK1/2， and RAS protein.

Intein is a functional protein that mediates self-cleavage from precursor protein and simultaneously connects the exteins on both sides of the intein via peptide bonds. Among all kinds of inteins, split intein has a wide range of applications in the field of antibody ligation. However, since the naturally split intein specifically recognizes the first three amino acid sequences “cysteine, phenylalanine, and asparagine”(CFN)of the extein, exogenous amino acids are inevitably introduced after the protein splicing reaction. In this study, the amino acid sequence “cysteine, aspartic acid, and lysine”(CDK)of the antibody hinge region was substituted for “CFN” as the recognition site for the intein and the split intein Npu DnaE was mutated into Npu*GEP DnaE. The results showed that the mutant could recognize “CDK” and the intein splicing reaction could successfully take place. The factors affecting the intein splicing reaction platform, such as pH, temperature and the concentration of NaCl and DTT were investigated in this study. The results showed that the splicing reaction of the mutant performed well, which indicated its potential usefulness in bispecific antibody assembly. In conclusion, the problem of introducing foreign amino acids was alleviated, the broadness of intein substrate was further expanded, and further technical support for the application of intein to the antibody assembly was provided.

Objective To observe the changes of iNOS, COX-2 and NF-κB in breast cancer MCF-7 cells treated with Yanghe decoction containing serum. Methods The MCF-7 human breast cancer cells were divided into blank group, low dose group, middle dose group and high dose group (4.5, 9, 18 g/kg, respectively). Real-time RT-PCR was used to detect the mRNA expression of iNOS and COX-2 at 24, 48, 72 and 96 hours, and Western-blot was used to detect the expression of NF-κB protein. Results Compared with the blank group, the expression of iNOS and COX-2 mRNA in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours(P<0.05), the the expression of NF-κB protein in the low dose group, middle dose group and high dose group decreased significantly at 24, 48, 72 and 96 hours (P<0.05). So it reduced with the increase of drug concentration and time. There was no significant difference in 72 hours after intervention. Conclusions The Yanghe decoction could reduce the expression of NF-κB, and then reducing the related inflammatory factors COX-2 and iNOS.

Objective: To investigate the effect of Yanghe decoction serum on the proliferation of breast cancer stem cells MDA-MB-231 and the expression of Smad3 and NF-κB. Methods: According to the random number table method, 20 female SD rats were divided into blank control group and Yanghe decoction high, medium and low dose group. Drug serum were given gastric gavage of Yanghe decoction 28, 14, 7 g/kg, and control group were given gastric gavage of same volume saline. Each group of rats was given orally for 3 days to prepare 10% Yanghe decoction serum and blank serum. MDA-MB-231 cells were divided into blank control group and Yanghe decoction low, medium and high dose group according to random number table method. Low, medium and high dose Yanghe decoction groups were cultured with medium containing 10% Yanghe decoction high, medium and low dose drug serum, and the blank control group was cultured with medium containing 10% control serum. After drug intervention, the effects of each group on the proliferation of MDA-MB-231 cells at 24 h, 48 h, and 72 h were detected by MTT assay. The expression of Smad3 and NF-κB protein in each group was detected by Western blot. The expression of Smad3 and NF-κB mRNA in each group were detected by real-time fluorescence quantitative PCR. Results: Compared to the control group, Yanghe decoction low, medium and high dose group of cells for 24 h (1.143 ± 0.093, 0.953 ± 0.069, 0.874 ± 0.041 vs. 1.239 ± 0.160), 48 h (1.223 ± 0.129, 0.989 ± 0.093, 0.864 ± 0.105 vs. 1.389 ± 0.095), 72 h (1.092 ± 0.147, 0.881 ± 0.095, 0.719 ± 0.086 vs. 1.353 ± 0.150) proliferation activity were significantly lower (P<0.05 or P<0.01). After 48 h of drug intervention, the expression of Smad3 protein (0.974 ± 0.098, 0.844 ± 0.084, 0.789 ± 0.105 vs. 1.214 ± 0.012), NF- κB p50 protein (0.994 ± 0.047, 0.911 ± 0.015, 0.765 ± 0.084 vs. 1.147 ± 0.103) in the low, medium and high dose Yanghe decoction group were significantly lower than those in the control group (P<0.05 or P<0.01). The expression of Smad3 mRNA (1.223 ± 0.129, 0.989 ± 0.093, 0.864 ± 0.105 vs.1.389 ± 0.095), NF-κB mRNA (1.092 ± 0.147, 0.881 ± 0.095, 0.719 ± 0.086 vs. 1.353 ± 0.150) significantly decreased in the Yanghe decoction group (P<0.05 or P<0.01). Conclusions Yanghe decoction can inhibit the proliferation of breast cancer stem cell MDA-MB-231, and lower the expression of Smad3 and NF-κB.