OBJECTIVE To investigate the effect of arctigenin(ATG) on liver fibrosis in rats induced by carbon tetrachloride(CCl4)and to explore its underlying mechanism. METHODS Sprague-Dawley rats were randomly divided into six groups:vehicle,ATG 3.0 mg · kg-1 group,CCl4 model group,CCl4+ATG 1.0 and 3.0 mg·kg-1 groups,and CCl4+colchicine(COL)0.1 mg·kg-1(toxicity)group. Liver fibrosis was induced by subcutaneous injection of CCl4 in rats for 8 weeks. ATG and colchicine were administrated ig once a day starting from the fifth week after the CCl4 treatment for 4 weeks subsequent. At the end of the study,glutamic pyruvic transaminase (GPT),glutamic oxaloacetic transaminase(GOT),albumin(ALB),and total bilirubin (TBIL) as well as the contents of hydroxyproline (HYP) in liver tissues were measured. Histopathological changes were observed in the liver tissues using hematoxyline-eosin(HE)and Masson’s trichrome staining. The proliferation of hepatic stellate cells (HSC) and expression of cell cycle-related proteins were assayed by indirect immunofluores?cence staining and Western blotting,respectively. RESULTS Compared with CCl4 model group,ATG 1.0 and 3.0 mg · kg-1 improved the liver function by decreasing serum contents of GPT,GOT and TBIL (P<0.05),and increasing serum content of albumin(P<0.05). Histological results indicated that ATG 1.0 and 3.0 mg · kg-1 alleviated liver damage and reduced the formation of fibrous septa. Moreover, ATG 1.0 and 3.0 mg · kg-1 significantly decreased liver HYP when compared with CCl4 model group(P<0.05). In addition,CCl4-induced proliferation of activated HSC was inhibited by ATG 1.0 and 3.0 mg·kg-1, and this was accompanied by down-regulation of cyclin D1,cyclin-dependent kinase(CDK)2,CDK4, and proliferating cell nuclear antigen (PCNA)(P<0.05),and up-regulation of p27kip1 in activated HSC (P<0.05). CONCLUSION ATG can alleviate hepatic injury and fibrosis induced by CCl4,which is probably associated with suppression of the proliferation of activated HSC.

Objective To evaluate laparoscopic D2 lymph node dissection gastrectomy in the treatment of advanced gastric cancer.Methods The clinical data of 239 cases of advanced gastric cancer admitted from January 2004 to June 2011 were respectively analyzed,patients were divided into laparoscopic resection group and open surgery group.Data analysis was performed by SPSS 19.0 statistical software.Results There were 102 cases in laparoscopic group,and 137 cases in open group.The length of incision,operative blood loss,recovery time of gastrointestinal function,food-taking time and postoperative hospital stay in laparoscopic operation group were (5.0 ± 1.1) cm,(70 ± 44) ml,(57 ± 14) h,(68 ± 12) h,(7.1 ± 1.4) d and in open operation group were (17.4 ± 2.1) cm,(107 ± 59) ml,(75 ± 12) h,(91 ±15) h,(9.9 ± 1.8) d respectively.There were significant differences between the two groups (t =-58.86,-5.50,-10.72,-12.58,-12.58,all P =0.00).There was no significant differences between the two groups in operative time (t =1.63,P =0.11),with operative time in laparoscopic operation group of (192 ± 32) min,and (185 ± 30) min in open group.Average proximal,distal cutting edge and the average number of lymph node harvested were (5.0 ± 1.0) cm,(4.7 ± 0.8) cm,(27.6 ± 7.2) in laparoscopic operation group,and (5.1 ±0.9) cm,(4.7 ±0.9) cm,(27.0 ±6.5) in open group (t =-0.61,0.10,0.68,P ＞ 0.05).The 3-,5-,7 d white blood cell counts in laparoscopic group was (11.1 ± 1.3) ×109/L,(9.5 ± 1.4) × 109/L,(7.0 ± 1.5) × 109/L,and (12.8 ± 1.3) × 109/L,(11.1 ± 1.5) × 109/L,(8.6 ± 1.3) × 109/L,in open group (t =-9.83,-8.88,-9.40,all P =0.00).Complications developed in 9.8 ％ (10/102) in laparoscopic operation group,and 17.5 ％ (24/137) in open group (x2 =0.285,P =0.09).The 1-year,3-year,5-year survival rate of patients in laparoscopic group were 96.1％,74.1％,47.2％,and 95.6％,70.0％,50.9％ in open group (x2=0,0.04,0.21,P ＞0.05).Conclusions In selected cases,laparoscopic D2 lymph node dissection gastrectomy for advanced gastric cancer is safe and effective,and long-term outcomes are satisfactory.

OBJECTIVE: To evaluate the efficacy of personal sublingual immunotherapy (SLIT) with dermatophagoides to study the efficacy of dermatophagoides farinae drops for allergic rhinitis (AR) of different age groups. METHOD: The current study had analyzed the efficacy of SLIT in 150 patients with AR who were sensitized to house dust mites. All patients were treated with dermatophagoides farinae drops and combined with symptomatic treatment. The patients were divided into groups 1-5, group 1:17 patients (4-7 years old), group 2: 38 patients (> 7-12 years old), group 3:31 patients (> 12-18 years old), group 4: 38 patients (> 18 - 40 years old), group 5: 26 patients (> 40-63 years old). The total nasal symptom scores (TNSS) and total medicine scores (TMS) were recorded at each visit. Before and after SLIT for 0.5 year, 1 year and 1.5 to 2.0 years, the TNSS and TMS of each patient were evaluated. The dosage adjustment of immunotherapy according to the patient's symptoms were performed. RESULT: The TNSS and TMS had continuously improved significantly after SLIT for half year, 1 year and 1.5 to 2.0 years in all groups as compared with baseline (P < 0.05). There were no significant differences in the different age groups for TNSS and TMS during all time points. CONCLUSION Individualized SLIT with dermatophagoides farinae drops for 1.5-2.0 years is the most effective in the patients with allergic rhinitis of different age groups. And equivalent efficacy could be achieved for different age groups.
Adolescent ; Adult ; Age Factors ; Animals ; Antigens, Dermatophagoides ; administration & dosage ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; Rhinitis, Allergic ; therapy ; Sublingual Immunotherapy ; Treatment Outcome ; Young Adult

OBJECTIVE: To evaluate the efficacy of sublingual immunotherapy (SLIT) with Dermatophagoides farinae drops for allergic rhinitis (AR) of different symptom severity. METHOD: This retrospective analysis to receive SLIT treatment of 143 cases of patients with allergic rhinitis, according to the severity of disease symptoms divid- ed into two groups, moderate group (62 patients) and severe group (81 patients). Before SLIT and after SLIT for half year, 1 year and 1. 5-2.0 years, the TNSS, TMS and sign scores of patients with allergic rhinitis were evaluated. RESULT: The TNSS, TMS and sign scores had continuously improved significantly after SLIT for half year, 1 year and 1.5-2.0 years in two groups as compared with baseline (P < 0.05). Before SLIT, TNSS and sign scores of severe group had a significantly higher level than moderate group (Z = 10.40, 2.40, P < 0.05), while TMS of two groups had no significant differences (Z = 0.00, P > 0.05). Half year after SLIT treatment, in two groups for sign scores, there were significant differences (Z = 3.32, P < 0.05), and there were no significant differences for TNSS (Z = 1.58, P > 0.05) and TMS (Z = 0.37, P > 0.05). 1 and 1.5-2.0 years after SLIT, there were no significant differences in two groups for TNSS, TMS and sign scores (P > 0.05). CONCLUSION SLIT with Dermatophagoides farinae drops for 1.5-2.0 years is effective in the patients with allergic rhinitis of different symptom severity. And equivalent efficacy could be achieved for different symptom severity.
Administration, Sublingual ; Animals ; Antigens, Dermatophagoides ; administration & dosage ; Dermatophagoides farinae ; Humans ; Retrospective Studies ; Rhinitis, Allergic, Perennial ; drug therapy ; Sublingual Immunotherapy ; Treatment Outcome

OBJECTIVE： To establish HPLC fingerprint of Zhuang and Yao medicine Lespedeza formosa， and to provide reference for quality control of L. formosa from different producing areas. METHODS： Totally 10 batches of samples were collected from 5 producing areas as Guangxi Nanning， Guilin， Wuzhou and so on. HPLC fingerprints was established and similarity analysis was carried out by using “Similarity evaluation system of TCM chromatographic fingerprint” （2012 edition） software. The determination was performed on Inertsil ODS-3 column with mobile phase consisted of acetonitrile-0.2％ phosphoric acid solution （gradient elution） at the flow rate of 0.8 mL/min. The detection wavelength was set at 350 nm， and the column temperature was 35 ℃. The sample size was 10 μL. Common peaks were identified by comparing substance control. Cluster analysis and principal compoent analysis were performed by using IBM SPSS 21.0 statistical software. RESULTS： HPLC fingerprint was established， and 12 common peaks were calibrated. 3 common peaks were identified （common peak 8， 10， 11 were chafotalin， vitexin and isovitexin）. The similarity of 10 batches of samples were all higher than 0.9. Through cluster analysis， 10 batches of medicinal materials could be clutered into 2 groups. According to the principal component analysis， the cumulative variance contribution rate of the two principal component factors was 86.108％ （contribution rates of first principal components and second principal components were 66.891％ and 19.217％）. CONCLUSIONS： HPLC fingerprint of L. formosa is established successfully. The method is simple and easy to use， provides a reliable evalution method for quality control of L. formosa.

ObjectiveTo investigate the effect of Stemona tuberosa alkaloids （STA） on apoptosis and phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） and c-Jun N-terminal kinase/p38 mitogen-activated protein kinase （JNK/p38 MAPK） signaling pathways in human lung cancer A549 cells. MethodA549 cells were classified into blank group and STA groups （100， 150， 200， 250， 300 mg⋅L^-1）. Thiazole blue （MTT） assay and colony formation assay were used to evaluate the proliferation of A549 cells. Apoptosis was observed based on Hoechst 33258 staining， flow cytometry， and Annexin V-FITC/PI staining. Western blot was employed to detect the expression of apoptosis-related proteins cysteine-aspartic acid protease-3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， and Bcl-2， and the expression of PI3K， phosphorylated （p）-PI3K， Akt， p-Akt， JNK， p-JNK， p38 MAPK， and p-p38 MAPK. ResultCompared with the blank group， STA groups （150， 200， 250， 300 mg⋅L^-1） demonstrated the increase in inhibition rate of cell proliferation （P<0.01） and cell clone inhibition rate， and decrease in cell clone formation rate （P<0.01）. In comparison with the blank group， STA groups showed typical characteristics of apoptosis， such as chromatin condensation and enhanced fluorescence reaction. The apoptosis rate of STA groups was significantly higher than that of the blank group （P<0.01）. Compared with the blank group， STA （150， 200， 250， 300 mg⋅L^-1） significantly up-regulated the protein expression of Caspase-3 and Bax （P<0.05， P<0.01） and down-regulated the expression of Bcl-2 protein （P<0.01）. Compared with the blank group， STA had no significant influence on the total protein expression of PI3K， Akt， JNK， and p38 MAPK. However， STA （150， 200， 250， 300 mg⋅L^-1） significantly decreased the levels of p-PI3K and p-Akt （P<0.05， P<0.01） and increased the level of p-p38 MAPK （P<0.05， P<0.01）. Compared with the blank group， STA （200， 250， 300 mg⋅L^-1） significantly raised the level of p-JNK （P<0.05， P<0.01）. ConclusionSTA can inhibit the proliferation and induce the apoptosis of A549 cells by inhibiting PI3K/Akt signaling pathway and activating JNK/p38 MAPK signaling pathway.

Objective : To investigate the effects of DNA demethylation drugs combined with histone deacetylase inhibitors on fragile X mental retardation 1 neighbor protein (FMR1NB) expression and its promoter methylation in human oral cancer cells and try to find a strategy of weakening the heterogeneity of FMR1NB expression . Methods: Human oral cancer cell lines C al27 and SCC⁃9 were treated with decitabine (DAC) , an inhibitor of DNA methyltransferase , combined with trichostatin A ( TSA) and valproic acid ( VPA) , inhibitors of histone deacetylase . Then reverse transcription⁃polymerase chain reaction ( RT⁃PCR) , quantitative real ⁃time PCR ( qRT⁃PCR) and Western blot were used to detect the expression of FMR1NB and pyrosequencing was used to detect the methylation of FMR1NB promoter. Results : Compared with the blank control group , DAC and its combination with TSA and VPA significantly induced the expression of FMR1NB mRNA and protein in C al27 and SCC⁃9 cells . Compared with DAC alone group , FMR1NB mRNA expression of each DAC⁃combined drug groups significantly increased , but FMR1NB protein did not significantly change in C al27 cells; for SCC⁃9 cells , except for DAC + TSA group , the mRNA and protein levels of FMR1NB significantly increased in all other groups . In addition , there was no significant difference in the expression of FMR1NB mRNA and protein between the three⁃combined drugs group and two-combined drugs groups . Further methylation assay showed that the methylation level of the overall FMR1NB promoter and its each CpG site measured were reduced to varying degrees in all treatment groups except for three⁃combination drug group of SCC⁃9 . Conclusion DAC and its combination with TSA and VPA can enhance the expression of FMR1NB by mediating the demethylation of FMR1NB promoter , wherein the enhanced expression effect of the combination of the two drugs is stronger , suggesting that they have the potential to weaken the heterogeneity of FMR1NB expression and improve the immunotherapy effect of oral cancer.

Objective: To study the expression of long non-coding RNA LINC01152 in glioma and its influence on the malignant biological behavior of glioma cells． Methods: LINC01152 expression in glioma was analyzed by LncSpA V2．0 and GEPIA database．qRT-PCR was applied to detect the expression of LINC01152 mRNA in 10 samples of human normal brain tissues，40 samples of glioma tissues and 5 glioma cell lines．GO and KEGG enrichment analysis of LINC01152 co-expressed genes were performed using the DAVID database to predict the related functions． The AnoLnc2，TargetScan，LinkedOmics and miRDB databases were used to predict the LINC01152 related miRNAs and target genes to construct a ceRNA regulatory network．LINC01152 expression was knocked down in glioma cell lines by small interfering RNA (siRNA) transfection．The CCK-8 test，scratch healing experiments，Transwell，flow cytometry and Western blot experiments were used to measure the influence of LINC01152 on the proliferation，migra- tion，invasion and apoptosis of glioma cells. Results : Database analysis showed that compared with other tumor types，LINC01152 was highly expressed in glioblastoma (GBM) and low grade glioma (LGG) ，and was higher than normal brain tissue．qRT-PCR showed that the expression of LINC01152 mRNA in glioma tissues was significantly higher than that in normal brain tissues (P＜0. 01)．The expression of LINC01152 was correlated with Ki-67 (P < 0. 05) ，but not with clinical parameters such as gender，age，tumor size，P53 protein，glial fibrillary acidic protein (GFAP) ，O-6-methylguanine-DNAmethyltransferase ( MGMT) and WHO grade of glioma patients． Functional enrichment analysis of co-expressed genes indicated that the LINC01152 was mainly involved in biological processes such as cell adhesion and synaptic signaling．LINC01152-miRNA-mRNA regulatory network was constructed according to predicted target genes．After down-regulation of LINC01152 expression，the proliferation，migration and invasion abilities of A172 and U87 cells decreased(P＜0. 01) ，while the apoptosis of glioma cells significantly increased (P＜0. 001) ． Conclusion LINC01152 is highly expressed in glioma，which can promote the malignant biological behavior of glioma cells by enhancing proliferation，migration as well as invasion and inhibition of apoptosis.