AIM:To investigate the effect of AG 490 on the expression of VEGF and HIF-1α, and the capacity of invasion in human erythroleukemia (HEL) cells.METHODS:The HEL cells were treated with AG490 at different con-centrations .The cell viability was detected by CCK-8 assay.The apoptosis was detected by Hoechst staining .The apoptosis and the cell cycle were analyzed by flow cytometry .The capacity of migration was evaluated by Transwell assay .The mRNA expression level of JAK2 was measured by RT-PCR.The protein levels of p-JAK2, VEGF and HIF-1αwere determined by Western blot.RESULTS:The HEL cell viabilities were 88%, 75%, 48%, 10%and 0.12%after treated with AG490 at 20, 40, 60, 80 and 100 μmol/L for 48 h, respectively.The results of Hoechst staining showed that brilliant blue cells in 80 μmol/L AG490 group was significantly increased compared with control group for 48 h.The apoptosis rate of 80μmol/L AG490 group was significantly increased compared with control group at 48 h after AG490 treatment.The number of membrane-permeating HEL cells in 20μmol/L AG490 group at 24 h after AG490 treatment was significantly lower than that in control group (P<0.05).The mRNA level of JAK2 decreased in a concentration-dependent manner after the HEL cells were treated with different concentrations of AG 490 for 48 h.The protein levels of p-JAK2, VEGF and HIF-1αwere lower in AG490 treatment groups than those in control group (P<0.05).CONCLUSION: AG490 inhibits the expression of VEGF and HIF-1αin HEL cells by inhibiting JAK2 pathway.

Long noncoding RNA (lncRNA) is a class of RNA molecules that longer than 200 nucleotides.lncRNA is lack of functional open reading frames and has no protein coding ability.Recently, accumulating evidences indicate that lncRNA plays active roles in tumor carcinogenesis and progression.The aberrant expression of lncRNA is significantly correlated with the growth,metastasis,and therapy-resistance of digestive system tumors.Thus,lncRNA may be served as new targets for the diagnosis,treatment,and progno-sis of patients with digestive system tumors.

ObjectiveTo investigate the relationship between metastasis-associated gene 1 ( MTA1 )expression and invasive and metastatic ability of cervical cancer cell. MethodsThree kinds of plasmids pcDNA3( control group), pcDNA3-MTA1 ( MTA1 group) and pSilencer3. 1-MTA1-siRNA ( MTA1-siRNAgroup) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability. ResultsCompared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P＜0. 05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ±6, 169 ± 10 and 57 ±5,respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group(P ＜ 0. 05 ). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69. 3 ± 3. 6) ％, ( 80. 4 ± 5. 6 ) ％ and ( 39. 2 ± 7.4 ) ％ separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P ＜0. 05 ). In the migration assay, the number of cells migrated to the bottom side of the membrane in control,MTA1 and MTA1-siRNA group were 153 ± 17,247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19,354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA 1-siRNA group showed lower cell migration and invasion ability (P ＜ 0. 05 ). In cell cycle experiment, no significant differences of cell proportions including G1, S and G2 stage were found among three groups (P ＞ 0.05).In animal experiment, compared with control group,MTA1 group showed accelersted tumor formation and growth,whilethe MTA1-siRNA group showed the reverse effect ( P ＜ 0. 05 ). ConclusionsMTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

AlM:To study the treatment effect of traditional Chinese medicine in the treatment of high myopia macular hemorrhage, using Chinese medicine etiology and pathogenesis, syndrome differentiation treatment, and provide the basis for the clinical treatment. METHODS: Eighty patients ( 135 eyes ) with high myopia macular hemorrhage were selected in the hospital from January 2012 to september 2014 as treatment group, and applied traditional Chinese medicine treatment. Forty-five patients (64 eyes) with the same period, as the control group, received routine western medicine treatment. After 1mo treatement, the treatment effect and vision improvement situation of two groups were observed, and after 6mo follow-up, the relapse was observed.RESULTS: The total effective rate of treatment group was 85. 19% (115/135), higher than the control group 78. 13% (50/64) (P<0. 05). The average corrected visual acuity of treatment group was 0. 48±0. 11, higher than the control group 0. 36 ± 0. 09, the difference was statistically significant (P<0. 05). The average diopter and macular bleeding scope of the treatment group were -9. 81±0. 85D and 0. 51 ± 0. 27PD, lower than the control group -10. 76 ± 0. 91D and 0. 78 ± 0. 23PD, the differences were statistically significant ( P < 0. 05 ). The eye ground hemorrhage absorption time of treatment group was 25. 34±2. 28d, less than the control group 29. 72 ± 2. 13d, the difference was statistically significant (P<0. 05). The bleeding again of the control group 7. 81% ( 5/64 ), higher than the treatment group was 5. 19% (7/135), the difference was statistically significant (P<0. 05).CONCLUSlON: Evidence-based treatment of traditional Chinese medicine for high myopia macular hemorrhage has good clinical effect, can shorten the treatment time, and is beneficial to the recovery of postoperative vision, worthy of clinical popularization and application.

Objective:To investigate the role of service-learning model in medical ethics education. Meth-ods:A qualitative and quantitative research was conducted within 831 undergraduates and 302 graduates in Shan-tou University Medical College. Results: Service-learning model was well accepted by most medical students. Higher medical ethics awareness and more positive tendency of behavior choices were found on the group who par-ticipated in serving-learning model activities, which also revealed a positive correlation relationship to participated frequency. Conclusions:Serving-learning model plays an important role in medical ethics education, which is an effective way to deeply practice Opinions on Deepening the Medical and Educational Reform toward Personnel Training in Clinical Medicine.

OBJECTIVETo explore the effects of principal-subordinate acupoints combination on improving myocardial ischemia, and the gene regulatory pathways for the protection of myocardial ischemia.

METHODSAccording to the random number table method, 70 SPF Wistar male rats were divided into a normal group, a model group, a LY294002 group, an insulin-like growth factors-1(IGF-1) group, a Neiguan group, an acupoint combination group and an acupoint combination + LY294002 group, 10 rats in each one. Rats in the normal group were injected with 0.9% NaCl solution, while rats in the remaining groups were treated with abdominal subcutaneous injection of isoroterenol hydrochloride to establish the rat model of myocardial ischemia. Rats in the LY294002 group and IGF-1 group were treated with injection of LY294002 solution and IGF-1 solution for 14 days. Rats in the Neiguan group were treated with electroacupuncture (EA) at "Neiguan" (PC 6) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination group were treated with EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) by using Han-200 EA apparatus for 10 min per treatment. Rats in the acupoint combination + LY294002 group were treated with LY294002 solution for 14 days, and EA at "Neiguan" (PC 6), "Zusanli" (ST 36) and "Guanyuan" (CV 4) was given before model establishment, once a day for 21 days. EA pretreatment was given before model establishment in all acupuncture groups. The heart rate (HR) and ST segment voltage were detected before and after treatment; the myocardial pathological morphology was observed by HE staining; the expressions of P13K mRNA and Akt mRNA were tested.

RESULTSAfter modeling, HR and ST segment voltage in all intervention groups were higher than those in the normal group (all P < 0.01); after the intervention, the HR and the ST segment voltage in the acupoint combination group, IGF-1 group and IGF-1 group were improved (P < 0.01, P < 0.05), which was more significant in the acupoint combination group and Neiguan group (all P < 0.01). As for the myocardial pathological morphology, obvious myocardial ischemia was observed in the model group, and that in the LY294002 group was the most serious, and that in the acupoint combination+ LY294002 group was moderate. After intervention, the myocardial pathological damage in the IGF-1 group, Neiguan group and acupoint combination group was significant improved, which was more significant in the IGF-1 group and acupoint combination group. As for the expression of PI3K mRNA and Akt mRNA, compared with normal group, the expression of PI3K mRNA was increased in the remaining groups after modeling (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0. 01). The expression of Akt mRNA in the LY294002 group and acupoint combination + LY294002 group was reduced (P < 0. 01, P < 0.05), while that in the remaining groups was increased (P < 0.01, P < 0.05), which was more significant in the IGF-1 group and acupoint combination group (all P < 0.01).

CONCLUSIONThe principal-subordinate acupoints combination could improve heart rate and ST segment voltage in rats with chronic myocardial ischemia, reduce myocardial pathological damage, which is superior to single selection of "Neiguan" (PC 6). The PI3K/Akt signaling pathway may be involved in the regulation mechanism of principal-subordinate acupoints combination for the protection of chronic myocardial ischemia.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Chronic Disease ; therapy ; Electroacupuncture ; Electrocardiography ; Heart Rate ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Male ; Myocardial Ischemia ; enzymology ; pathology ; physiopathology ; therapy ; Myocardium ; pathology ; Phosphatidylinositol 3-Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Rats ; Rats, Wistar

This study aims to investigate whether the nucleoprotein (NP) of severe fever with thrombocytopenia syndrome virus (SFTSV) can impact the cellular immunity of host cells. Gene segments that encode the NP and non-structural protein (NSs) of SFTSV were inserted into eukaryotic expression vector VR1012. Host proteins that interact with NP and affect immunity were identified with co-immunoprecipitation (IP), SDS-PAGE, mass spectrometry (MS), and Western blot. Co-localization of NP and the identified host proteins was confirmed by confocal microscopy. A 60kD SSA/Ro, a protein related to immunity, interacted with NP, as found by IP and MS. Confocal microscopy showed that NP and SSA/Ro were co-localized in cytoplasm. These results indicated that SFTSV NP may specifically bind to 60kD SSA/Ro and cause a series of immune responses and clinical symptoms.
Bunyaviridae Infections ; genetics ; metabolism ; virology ; HEK293 Cells ; Humans ; Nucleoproteins ; genetics ; metabolism ; Phlebovirus ; genetics ; metabolism ; Protein Binding ; Ribonucleoproteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

Objective To investigate the expression levels of serum LINC00978 in gastric cancer patients and its correlation with clinicopathological characteristics,and evaluate its clinical diagnostic value.Methods The expression levels of LINC00978 in gastric cancer cells and serum samples from the patients with gastric cancer or gastric benign diseases and healthy controls were detected by real-time qRT-PCR,and its correlations with clinicopathological parameters were analyzed.The diagnostic efficacy of serum LINC00978 was evaluated by the receiver operating characteristic (ROC) curve.Results The expression levels of LINC00978 in MGC-803 cells were significantly higher than that in GES-1 cells (18.88 ± 1.15 vs 1.00 ± 0.03,t =-21.926,P ＜ 0.05).The expression levels (median[P25,P75]) of serum LINC00978 in gastric cancer patients (3.525 [1.385,8.954]) were significantly higher than those in the patients with gastric benign diseases (0.419 [0.258,1.369],Z =-4.834,P ＜ 0.01) and healthy controls (0.814 [0.351,2.510],Z =-4.686,P ＜ 0.01).The expression levels of serum LINC00978 in gastric cancer patients were significantly related to lymphatic metastasis (r =0.448,P ＜ 0.01),invasive depth (r =0.369,P ＜ 0.01) and TNM stage (r =0.383,P ＜ 0.01),and those after operation significantly lower than before operation (0.17 [0.15,0.39] vs 0.98 [0.59,1.61],Z =-5.731,P ＜ 0.01).There was no significant difference in serum LINC00978 levels between the patients with gastric benign diseases and healthy controls (Z =-1.693,P ＞ 0.05).The area under the ROC curve (AUCROC) and 95％ confidence interval (CI) of serum LINC00978 levels in the diagnosis of gastric cancer were 0.807 and 0.732-0.882,respectively.When the cutoff value was 1.806,the sensitivity and specificity for the diagnosis of gastric cancer were 71.4％ and 75.9％,respectively.Conclusion The expression levels of serum LINC00978 in gastric cancer patients increase and are related to lymphatic metastasis,invasive depth and TNM stage,which may be served as a novel biomarker for the diagnosis of gastric cancer.

To evaluate the adjuvant effect of recombinant enterovirus 71 (EV71) subunit vaccine formulated with chitosan, rabbits were orally immunized with recombinant VP1 (rVP1) or rVP1 mixed with chitosan adjuvant. Levels of virus-specific IgG and IgA antibodies in sera, mucosal wash buffer (intestine, nasal cavity, and lung), and feces were determined by indirect enzyme-linked immunosorbent assay (ELISA). The titers of neutralizing antibodies against EV71 were determined using cytopathic effect-based neutralizing assay, and levels of cytokines (IFN-gamma and IL-4) secreted from in vitro-cultured rabbit splenic lymphocytes under antigen stimulation were also determined by ELISA. Results showed that immunization with rVP1 alone could only induce low levels of serum IgG and mucosal IgA, while rVP1 combined with chitosan adjuvant were able to induce significantly higher levels of antibodies, rVP1 can only induce neutralizing antibodies when used in combination with chitosan. Levels of IFN-gamma and IL-4 in the group immunized with rVP1 plus chitosan were significantly higher than those in the group immunized with rVP1 only or those in the control groups. Our study lays the foundation for development of oral VP1 vaccine against EV71 infection.
Adjuvants, Immunologic ; administration & dosage ; Animals ; Antibodies, Viral ; immunology ; Chitosan ; administration & dosage ; immunology ; Enterovirus A, Human ; genetics ; immunology ; Enterovirus Infections ; immunology ; prevention & control ; virology ; Female ; Humans ; Rabbits ; Vaccination ; Vaccines, Subunit ; administration & dosage ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology ; Viral Vaccines ; administration & dosage ; genetics ; immunology

OBJECTIVETo investigate the plasma levels of ascorbic acid and vitamin E in patients with liver cirrhosis and to explore their significance.

METHODSThe plasma levels of ascorbic acid,vitamin E and lipoperoxides in patients with liver cirrhosis were measured, and the results were compared with those of sex-and age-matched healthy subjects.

RESULTThe plasma levels of ascorbic acid, vitamin E and lipoperoxides in the patients group were (42.94 +/-6.99)micromol/L, (17.99 +/-3.51)micromol/L and (14.09 +/-1.28)micromol/L, respectively, while those in the control group were (53.30 +/-9.45)micromol/L (t=9.50, P=0.000), (24.59 +/-7.22)micromol/L (t=7.94, P=0.000) and (12.11 +/-1.20)micromol/L (t=17.21, P=0.000), respectively.

CONCLUSIONThe levels of ascorbic acid and vitamin E in patients with liver cirrhosis decrease significantly,which may indicates the disturbance of balance between oxidation and antioxidation.

Adult ; Aged ; Aged, 80 and over ; Ascorbic Acid ; blood ; Female ; Humans ; Lipid Peroxides ; blood ; Liver Cirrhosis ; blood ; metabolism ; Male ; Middle Aged ; Vitamin E ; blood