Objective To investigate paclitaxel and ezrin-siRNA therapy for metastatic liver cancer.Methods Human metastatic liver cancer cells MHCC97-H were divided into four groups in vitro and in vivo,including control cells,exposure to paclitaxel alone,ezrin-siRNA alone,and the combination of paclitaxel and ezrin-siRNA.Nude mice lung lesions were formed by tail vein injection of tumor cells.The tumor cells'apoptosis rate,wound healing ability,the ability to cross an artificial membrane,and the numbers of metastatic lung lesions were measured.Results The apoptosis rate in normally cultured MHCC97-H tumor cells was 2.52％,compared to the other three groups at 11.66％,2.35％,7.38％ respectively.The wound healing ability was best in the negative control and weakest in cells treated with combination paclitaxel and ezrin-siRNA.The numbers of cells which could cross the artificial membrane in the negative,paclitaxel,ezrin-siRNA,and combination paclitaxel and ezrin-siRNA group were 52.5 ± 3.9,32.2 ± 3.2,26.6 ± 2.4,and 19.4 ± 1.1 respectively.The numbers of metastatic lung lesions in the nude mice (n =40) were 12.5 ±2.4,8.2 ± 1.5,7.0 ± 1.3,and 3.8 ± 0.3 respectively.Conclusions The ezrin-siRNA combined with paclitaxel therapy demonstrated a greater efficacy over single agent therapy for depressing liver cancer growth,motility,aggression,and metastasis in vitro and in vivo.

Objective To study effect of the adipose tissue extract of greater omentum on human fibroblasts. Method The effect of the adipose tissue extract of greater omentum on human fibroblasts was observed by inverted microscope, MTT, flow cytometry, transmission electron microscope. Results The growth of fibroblasts was inhibited by the adipose tissue extract of greater omenturn. The apoptosis of fibroblasts was not inhibited, and the cell function of fibroblasts was inhibited and the mitosis of fibroblasts was affected. Conclusions The growth of fibroblasts can be inhibited by the adipose tissue extract of greater omentum. The formation and development of biliary cicatricial constriction may be affected by the adipose tissue extract of greater omentum.

Objective Via targeted inhibition of oncogene Bmi-1 expression by RNAi interfering technology in vitro, to observe its effect on the proliferation and cell cycle of gallbladder cancer cells. Methods Four miRNABmi-1 recombinant plasmids were constructed according to different Bmi-1 sites. RT-PCR and Western blot were used to mRNA and protein expression of Bmi-1 in gallbladder cancer cells were measured by RT-PCR and Western blot. mRNA and protein expression of Bmi-1 in gallbladder cancer cells. The most effective interfering plasmids in the miRNABmi-1 groups were transfected into GBC-SD cells. Cell proliferation and cell cycle were analyzed 48 h after transfection by BrdU and flow cytometry. Results Bmi-1mRNA expression in miRNAbmi1-1,-3 and-4 was significantly lower than the control group (P<0.05);and Bmi-1 protein expression in miRNAbmi1-2,-3 and-4 was significantly lower than the control group (P<0.05). The recombinant plasmid in miRNAbmi1-4, with the strongest inhibitive effect of Bmi-1mRNA and protein expression, was transfected into GBC-SD cells,then the cell proliferation rate (46.63 ± 5.31) was significantly lower in mRNABmi1-4 group than the control groups (P<0.05);G0/G1 phase cells increased (72.20 ± 1.71) and G2/M and S phase cells decreased (18.30 ± 7.21, 9.50 ± 6.01) in miRNABmi1-4 group. Both were significantly different from the control groups (P<0.05). Conclusions Targeting and silencing Bmi-1 expression can effectively inhibit the proliferation of GBC-SD cells and restrain the cell cycle atin G0/G1 phase. Bmi-1 gene may be a novel target for geneic therapy of gallbladder carcinoma.

Objective To investigate the diagnosis and surgical treatment of chronic acalculous cholecystitis characterized by absence of gallbladder wall contractability. Methods The clinical data of 42 patients with chronic acalculous cholecystitis in our hospital from January 2006 to December 2008were analysed. The patients were grouped into two groups: laparoscopic cholecystectomy (LC) group in 20 and non-surgical group in 22. The patients' symptoms on follow-up in the two groups were compared. Results The 42 patients with chronic acalculous cholecystitis were diagnosed by symptoms,ultrasound, fatty meal gallbladder contractability studies under ultrasound, fiber optic gastroscopy and magnetic resonance cholangiopancreatography (MRCP). In all patients, there was a complete absence of gallbladder wall contractability. In the LC groups, 20 patients received LC. 18 patients were followed up, and there were no symptoms. Two patients were lost to follow up. In the non-surgical group, 22 patients received non-surgical treatment. In 21 patients who were followed up, 19 patients had symptoms. One patient was lost to follow up. There was a significant difference between the LC group and the non-surgical group (P＜0.05). Conclusions Chronic acalculous cholecystitis characterized by absence of gallbladder wall contractability could be diagnosed by symptoms, ultrasound, fatty meal gallbladder contractability studies under untrasound, and MRCP. The optimal treatment of chronic acalculous cholecystitis characterized by absence of gallbladder wall contractability is LC.

Objective To explore the effect and mechanism of fragile site WWOX gene on regulating proliferation of gallbladder cancer cells in vitro. Methods The pcDNA3.0 - WWOX recombinant plasmid which was previous successfully built was transfected to GBC-SD cells and empty carrier by liposome medium. Liposome and GBC-SD were served as the negative control and the blank control,respectively. After 48 hours transfection, inverted microscope was used to observe the changes of gallbladder cancer cells' morphology,MTT and BrdU were used to detect the proliferation level of gallbladder cancer cells,and flow cytometry instrument was used to detect the change of the cell proliferation cycle. Results The results of inverted microscope shown: the number of GBC-SD cells in pcDNA3.0-WWOX group decreased significantly,the suspension cells and cell debris increased,while cells in the vector control,NC and Mock groups were in normal proliferation state. MTT test showed the proliferation levels of GBC-SD cells in pcDNA3.0-WWOX group was lower than those in the control group in 24 h,48 h,72 h,96 h and 120 h,and the differences were statistically significant(P < 0.05). The cell proliferation activity in the pcDNA3.0-WWOX group was obviously inhibited over time. BrdU detection results showed the cell proliferation rate of pcDNA3.0 - WWOX group was(0.44±0.03),while that in the three control groups was(0.78±0.02), (0.81±0.01)and(0.85±0.01),respectively. It showed that cell proliferation activity in pcDNA3.0-WWOX group was lower than the control groups,and the difference was statistically significant(P < 0.05). Cell cycle detection showed the cells increased in G0/G1 phase and decreased in G2/M and S phases of pcDNA3.0-WWOX group. The cell apoptosis rate was significantly higher and the proliferation index was significantly lower than those of the control groups(P < 0.05). However,there were no significant differences among the three control groups(P > 0.05). Conclusion The overexpression of WWOX gene in vitro could effectively inhibit the proliferation activity of gallbladder cancer cells. WWOX might participate in the development of the malignant biological behavior of gallbladder cancer cells. It is expected to become a new potential target for the gene therapy to gallbladder cancer.

The distribution of the Na-K-2Cl co-transporter (NKCC1) in the cochlear K+ cycling pathway in cochlea and cochlear histological changes in the NKCC1 knockout mice were investigated. By using immunohistochemistry and toluidine blue staining, the localization of NKCC1 in cochlea of the C57BL/6J mice and the cochlear histological changes in the NKCC1 knockout mice were observed. It was found that the NKCC1 was expressed mainly in the stria marginal cells and the fibrocytes in the inferior portion of the spiral ligament in the adult C57BL/6J mice. Subpopulation of the fibrocytes in the suprastrial region and the limbus was also moderately immunoreactive. While in the cochlea of the NKCC1 knockout mice, Reissner's membrane was collapsed and scala media disappeared, accompanied with the loss of inner hair cells, outer hair cells and the support cells. The tunnel of Corti was often absent. All the findings suggested the localization of NKCC1 in the cochlea was closely correlated with cochlear K+ cycling. Loss of NKCC1 led to the destruction of the cochlear structures, and subsequently influenced the physiological function of cochlea.

Objective To investigate the expression of HSPA9 in hepatocellular carcinoma(HCC) and its relationship with clinicopathological features and prognosis.Methods Forty-nine cases HCC treated by operative resection and follow up data in our hospital from January 2006 to January 2010 were retrospectively analyzed.Immunohistochemistry was performed to determine the expression of HSPA9 in HCC and paratumor tissues.The relationship between HSPA9 expression and clinicopathological features and prognosis was statistically analyzed.Results The HSPA9 protein expression in tumor tissue was higher that that in the paratumor tissue(t=6.601,P＜0.01),moreover the over expression of HSPA9 was significantly correlated with lymph node metastasis (P =0.005),TNM-stage(P =0.015),tumor differentiation (P =0.033),microvascular invasion (P =0.009) and recurrence (P =0.047).In the survival analysis results,the patients with over expression of HSPA9 had a much lower total survival rate(P=0.002)and much higher postoperative cumulative recurrence rate(P =0.003).There were significant differences in TNM-stage,microvascular invasion,lymph node metastasis,tumor differentiation and HSPA9 staining for overall survival and cumulative recurrence rate based on a univariate analysis(P＜0.05).Conclusion HSPA9 has over expression in HCC.The over expression of HS-PA9 is closely related to invasion and metastasis pathological features and can serve as an independent prognostic risk factor for predicting the prognosis of HCC.

Objective To establish the method of combined hepatocyte growth factor (HGF) with epidermal growth factor (EGF) cultured human gallbladder epithelial cells(HGBECs) in vitro .Methods The epithelial layer was peeled away from human gallbladder ,epithelial layer were digested with collagenase Ⅳ and scraped repeatedly .HGBECs were isolated and seeded in cell cul-ture plates containing medium supplemented with or without 10 ng/mL EGF or with 10 ng/mL HGF and 10 ng/mL EGF respec-tively .Then the morphologic changes of the cells were observed and taken photos with inverted phase contrast microscope ,and counted number of cells ,MTT assay detected vigor of cells in different groups .Results The number of the HGBECs of the HGF+EGF group was obviously more than the EGF group ,the duration of the HGBECs of the HGF+ EGF group was obviously longer than the EGF group(19 .3 ± 2 .5)d vs .(14 .2 ± 2 .4)d ,P< 0 .05 .And the HGBECs of the group with HGF+ EGF had better cell vigor .Conclusion HGF combines with EGF added to medium can obviously promote the proliferation of HGBECs and prolong the duration and stabilize morphology of HGBECs in vitro .

AIM:We generated transgenic mice of NKCC1-/-(homozygous mutant),NKCC1+/-(heterozygous)and NKCC1+/+(wild-type)that have a targeted disruption in the NKCC1 gene to investigate the role of Na-K-2Cl(NKCC1)channel in auditory function of the inner ear.METHODS:Hearing threshold and endocochlear potential(EP)were measured in the NKCC1-/-,NKCC1+/-and NKCC1+/+ mice by auditory brainstem response(ABR)and EP recordings,respectively.The inner ears of the mice were removed and examined morphologically with the light microscope.RESULTS:The auditory function of NKCC1+/+ mice was normal,the mean value for ABR thresholds in response to click sound was [(23.13?3.78)dB,SPL],EP was(98?16)mV.The mean value for ABR thresholds to click sound was elevated in NKCC1+/-mice [(38.49?12.29)dB,SPL],relative to that significantly increased in NKCC1+/+ mice(P

Objective To observe the characteristics of different level of normal abdominal aorta’s time-flow,time-velocity curve and to investigate the hemodynamic factors in role of the occurrence and development of abdominal aorta’s disease.Methods PC-MRI scanning was performed on 60 normal volunteers T12/L1-L4/5 disc at the level of abdominal aorta,time-flow and time-veloci-ty curve were generated respectively.The characteristics of blood flow curves at different levels of abdominal aorta were observed. Results The time-flow,time-velocity curve of the normal abdominal aorta in different levels have 2nd phase (no diastolic reflux) and 3rd phase (diastolic reflux).Conclusion The characteristics of blood flow curves of different levels of abdominal aorta are dif-ferent.