Objective To investigate the effect of the balance between helper T lymphocyte (Th) 1 cytokines and Th2 cytokines in the serum of hepatic fibrosis rats induced by CCl4 on the expression of β-catenin in their livers. Methods Seventy-five rats were divided into control group,treatment group and model group. Hepatic fibrosis models were built by subcutaneous injection of CCl4in male Wistar rats, then rats in treatment group were treated with pentoxifylline(PTX) by intragastric administration. The model rats without PTX treatment served as treatment control. The serum levels of Th1 cytokines(interferon-γ, IFN-γ; tumor necrosis factor-α, TNF-α), Th2 cytokines (interleukin-4,IL-4;interleukin-6,IL-6) and type Ⅳ collagen were measured by enzyme-linked immunosorbent assay (ELISA). The expression and distribution of β-catenin in liver tissue and the percentage of area of type Ⅳ collagen accounted in the liver were analyzed by immunohistochemical analyses. The changes in serum cytokines were depicted by using One-way ANOVA, and the correlation between β-catenin expression and cytokines levels or hepatic fibrosis development was elucidated by Bivariate.Results The serum levels of Th2 cytokines in model group were significantly higher than those in treatment group, and even higher than those in control group (F=71. 260,91. 732, all P＜0.05).However, the levels of Th1 cytokines in model group were significantly lower than in treatment group while even lower than those in control group(F= 50. 420,10. 625, all P＜0.05). Serum cytokines came out to be Th2 polarized. Meanwhile, the percentage of type Ⅳ collagen area accounted in the liver,which was corresponding to the degree of liver fibrosis, was lower in the treatment group than the model group and even lower in control group (F=3 000,both P<0.05). β-catenin expression in the liver of model group was stronger than that in the treatment group, and even stronger than that in normal group (F=92. 030, both P＜0. 05). According to β-catenin immunohistochemical analyses based on patho-photos, livers from the model group showed dark color in both cell cytoplasm and nucleus, while the liver from the treatment group only showed weak color in cytoplasm and no straining color in nucleus and the liver from normal group showed colors neither in cytoplasm nor in the nucleus.The expression of β-eatenin had a positive correlation with either the level of Th2 cytokines in serum of rats (r=0. 560,P＜0.01) or the percentage of area of type Ⅳ collagen accounted in the liver of rats (r=0. 757, P＜0. 01). Conclusions Th2 polarization in serum cytokines can aggravate the development of liver fibrosis by inducing the expression of β-catenin, which in the livers of rats is a reflection of the degree of the liver fibrosis.

Objective To study impact of traditional Chinese medicine liquid combined with compression method on wound healing of anal fistula after towing line operation. Methods 120 patients with anal fistula were divided into observation group and control group by random number table including 60 cases respectively. Observation group were given liquid washing fistula combined with sand bags oppression in wound on the basis of conventional nursing; Control group were given the routine nursing care. Curative effect, pain, wound healing time, recurrence rate and degree of satisfaction of two groups were compared. Results Clinical curative rate of observation group was 98.33% (59/60), which was obviously higher than 85.00% (51/60) of control group, and there was statistically significant difference (χ2=6.981 8, P<0.01). Postoperative pain score at the 1st, 2nd, 3rd day of observation group was (2.98± 2.01), (2.97±1.94), (3.06±1.92) points, which was lower than (5.48±1.76), (5.52±1.91), (5.42±2.01) points (t=13.336 9, 13.514 8, 12.261 3, P < 0.01). Wound healing time of patients in observation group was (23.96±3.28) days, which was obviously less than (32.34±4.51) days of control group (t=6.285 1, P<0.01);Recurrence rate in observation group was 1.67%(1/60), which was obviously lower than 11.67%(7/60) of the control group, and the difference was statistically significant (χ2=4.821 4, P < 0.05). Conclusions Applying traditional Chinese medicine liquid combined with compression method in wound healing of anal fistula after towing line operation can promote wound healing, improve curative effect, prevent recurrence, and improve patients′satisfaction.

To explore the characteristic genomics of syndrome of liver-kidney yin deficiency in peripheral blood mononuclear cells of hepatocellular carcinoma (HCC) patients.

A mouse model for acute hepatitis B virus (HBV) infection was established by using the hydrodynamical injection of mouse tail vein, in which the immunocompetent BALB/c mice were hydrodynamically injected with a competent replication plasmid pAAV-HBV1.2 having 1.2 fold over-length of HBV DNA. On day 1, 2, 4, 6 and 8 after injection, the levels of HBsAg, HBeAg and HBV DNA in blood serum were detected by using ELISA and fluorogenic quantitative PCR assay (FQ-PCR). And on day 8. HBsAg and HBeAg in liver tissue were assayed by immunohistochemical staining. It was found that HBsAg in blood serum could be detected on day 1 after infection in 14 of 16 mice (85.7%) injected with pAVV-HBV1.2 by using ELISA assay and the peak levels of HBsAg and HBeAg were attained during the first day after injection and then it dropped down gradually up to day 8 following injection. The titer of HBV DNA in blood serum attained its peak on day 2 and maintained a high level later on. On day 8 after injection, its titer was 1.9×10~4 copies/mL. The percentage of HBcAg-positive hepatocytes and HBsAg-positive hepatocytes in liver tissues were 5% and 2% respectively. Thus, by using the hydrodynamic injection with the competent replication plasmid, a mouse model for acute HBV infection is successively developed.

Objective To investigate the mechanisms of metformin in inducing autophagy of gastric cancer MNK-45 cells.Methods Human gastric cancer MNK-45 cells in logarithmic growth phase were incubated in the culture plates,and were divided into the intervention group [gastric cancer MNK-45 cells were intervened by metformin at different concentrations (2,4,8,16,32,64 mmol/L) for 24,48,72 hours] and the control group (gastric cancer MNK-45 cells were cultured in the DMEM medium).The inhibition rate of gastric cancer MNK-45 cells was detected by MTT method.The IC50 value of metformin on gastric cancer MNK-45 cells was 17 mmol/L.Gastric cancer MNK-45 cells were intervened by metforrnin at 17 mmol/L for 48 hours in the experimental group.Gastric cancer MNK-45 cells in the control group were cultured in DMEM medium at 17 mmol/L for 48 hours.The apoptosis of the gastric cancer MNK-45 cells of the 2 groups were detected by flow cytometry.The mRNA expressions of Bax and Bcl-2 of the 2 groups were detected by RT-PCR.The protein expressions of type Ⅰ LC3b,type Ⅱ LC3b,beclinl,AKT,p-AKT,mTOR,p-mTOR,P70s6k,p-P70s6k of the 2 groups were detected by Western blot.The measurement data were presented as (x) s,and were analyzed using the one-way ANOVA or repeated measures ANOVA.Data of the 2 groups were compared using the t test.Results The inhibition rates of gastric cancer MNK-45 cells were 3.0％ ± 1.1％,8.6％ ± 1.7％,15.9％ ± 1.6％,26.1％ ± 3.4％,37.5％ ± 2.3％,49.7％± 3.6％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 24 hours,5.2％± 1.9％,10.4％±2.1％,26.9％± 1.6％,49.5％± 1.6％,59.1％±2.0％,82.1％±2.2％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 48 hours,and 9.5％ ± 2.2％,17.6％ ± 1.4％,30.6％ ± 2.6％,63.2％ ± 2.6％,78.9％ ± 1.4％,93.3％ ± 2.7％ after intervention by metformin at concentrations of 2,4,8,16,32,64 mmol/L for 72 hours.There were significant differences in the inhibition rates among the 6 groups at the same time points (F =155.174,728.229,743.826,P ＜ 0.05),and significant differences were also observed within the same group at different time points (F =39.420,58.692,166.125,30.383,117.517,311.642,P ＜ 0.05).The apoptosis rates of gastric cancer MNK-45 cells in the experimental group and the control group were 25.4％ ± 1.7％ and 6.9％ ± 0.5％,with significant difference between the 2 groups (t =18.378,P ＜0.05).The relative mRNA expressions of Bax/Bcl-2 mRNA in the experimental group and the control group were 1.88 ± 0.16 and 1.00 ± 0.00,with significant difference between the 2 groups (t =9.743,P ＜ 0.05).The relative protein expressions of LC3 Ⅱ/LC3 Ⅰ and beclin 1 in the gastric cancer MNK-45 cells were 1.65 ± 0.08 and 1.47 ± 0.06 in the experimental group and 0.79 ± 0.03 and 0.56 ± 0.06 in the control group,with significant difference between the 2 groups (t =18.023,18.283,P ＜ 0.05).The relative protein expressions of AKT and P70s6k in the gastric cancer MNK-45 cells were 0.80 ±0.14 and 0.97 t0.21 in the experimental group and 0.96 ±0.17 and 1.37 ±0.23 in the control group,with no significant difference between the 2 groups (t =2.103,1.699,P ＞0.05).The relative protein expressions of mTOR and p-mTOR were 0.58 ± 0.l 1 and 0.57 ±0.15 in the experimental group and 1.88 ±0.23 and 2.36 ±0.25 in the control group,with significant difference between the 2 groups (t =11.293,10.979,P ＜ 0.05).No p-AKT and p-P70s6k expression was detected in the experimental group,and the expressions of p-AKT and p-P70s6k in the control group were 1.00 ± 0.00 and 1.00 ± 0.00,respectively.Conclusions Metformin could induce autophagy,inhibit proliferation and promote apoptosis of gastric cancer MNK-45 cells.The mechanism may be associated with the inhibition of mTOR expression and the expression of mTOR downstream proteins p-P70s6k by mefformin,and then the autophagy of gastric cancer MNK-45 cells happens.

Objective To evaluate the capability of model for end-stage liver disease (MELD)combined with serum sodium (MELD- Na,MELDNa and MESO scores) in predicting the prognosis of patients with decompensated liver cirrhosis in 6 and 12 months.Methods One hundred and nineteen patients with decompensated liver cirrhosis and completed follow-up data were retrospectively studied.The MELD,MELD- Na,MELDNa and MESO scores were calculated according to the clinical data of each patient.Receiver operating characteristic (ROC) curve and the area under the curve (AUC) was used to measure the values of the four models in predicting the 6 and 12 months survival,and Z-test was used to compare their predictive values.Results MELD,MELD-Na,MELDNa and MESO scores were significantly different between patients who survived and those who died within 6 and 12 months follow-up.The AUC for the MELD- Na,MELDNa and MESO scores were all more than 0.8 in predicting 6 and 12 months survival.However the differences of the AUC between the MELD score and MELD-Na,MELDNa,MESO scores were not significant in predicting 6 and 12 months survival.Conclusion The model for MELD combined with serum sodium can accurately predict the prognosis of patients with decompensated liver cirrhosis in 6 and 12 months,while these scores are not superior to MELD score.

In recent years, tumor is a refractory disease occurring frequently which is the main cause of death. Surgery, radiotherapy and chemotherapy are the usual therapeutic tools. However,radiotherapy and chemotherapy have serious side-effects and surgery can not be used effectively when metastasis happened. Therefore, tumor-targeted therapy has developed as a better way to cure tumor. Development of research on the use of PEG-PLGA nanoparticles as drug carriers are reviewed in this article, furthermore, problems about that are analysed.

AIM:To investigate whether the cigarette smoke extract (CSE) causes senescence of C2C12 myo-blasts and the relationship between senescence and histone deacetylase 2(HDAC2).METHODS: Murine C2C12 cells were induced to differentiate into myoblasts.The HDAC2 activator and inhibitor were used to investigate the effects of CSE in the myoblasts on cell senescence and the expression of HDAC2.The expression of HDAC2 at mRNA and protein levels was determined by real-time PCR and Western blot, respectively, and the positive cell rate of β-galactosidase staining for cell senescence was also detected.RESULTS:The optimal concentration of CSE was 60 mL/L and the intervention time was 24 h.After the intervention of CSE, the positive cell rate ofβ-galactosidase staining was increased, accompanied with the reduction of HDAC2 expression at mRNA and protein levels.The expression of HDAC2 at mRNA and protein levels was increased by 4, 5, 6, 7-tetrabromobenzotriazole (TBB), accompanied with the reduction of positive cell rate ofβ-galacto-sidase staining.Furthermore, when HDAC2 expression at mRNA and protein levels was reduced by HDAC2 inhibitor valp-roic acid, the positive cell rate of β-galactosidase staining was increased.CONCLUSION: CSE promotes the senescence by reducing the expression of HDAC2 in C2C12 myoblasts.

Objective:To screen the optimal formula and technology of demethoxycurcumin nano-structured lipid carriers. Meth-ods:The encapsulation efficiency as the main investigation index, the single factor exploration and orthogonal design were used to study the main factors affecting the quality of the nanoparticles. The optimal formula and technology were obtained. Results:The optimized parameters were as follows:the ratio of drug to lipid materials was 1 ∶40, the ratio of liquid lipid to total lipid materials was 10%, the amount of surfactants was 4% and the amount of lecithin was 2%. The prepared nanoparticles were spheric and regular. The size dis-tribution of the nanoparticles was narrow with the average particle size of 110nm and PDI of 0. 199. Conclusion:The optimized formula and technology of demethoxycurcumin nano-structured lipid carriers are stable and practicable,which provide reference for the further research of demethoxycurcumin.

Objective To establish a simple and practical superparamagnetic immunochromatographic test strip for rapidly monitoring human serum level of CA72-4. Methods Water-soluble carboxylated super-paramagnetic nanoparticles were prepared with a modified one-step hydrothermal synthesis method. Magnetic probe was prepared by immobilizing specific antibody (mAb1) onto the surface of nanoparticles. Following with optimization and assembly of the test strip , we evaluated sensitivity , specificity , stability of this method for serum CA72-4 detection. Results The optimized test strip provided not only the qualitative results, but also the high sensitivity quantitative detection through stable magnetic signal. The detection limit was 0.83 IU/mL. One hundred clinical samples ( 70 positive and 30 negative ) were measured to assess these test strips with high sensitivity (99%) and high specificity (93%). The test strip and magnetic signal possessed high stability. Conclusion A rapid and quantitative detection of CA72-4 by the test strip was accomplished. This method is rapid, sensitive and quantitative, possessing great potential in large sample screening or in-home testing.