Objective To study the effects of 5-aza-2-deoxycytidine (ZdCyd), a methylation inhibitor, on the growth of bile duct cancer cell line QBC939 in vivo and in vitro. MethodMTT method and flow cytometry were used to detect the growth and apoptosis of QBC939 after being treated with different dosage of ZdCyd singly or in combination with other chemotherapy drugs. Nude mouse model was used to test the effect of ZdCyd. Results5-aza-2-deoxycytidine blocks cell cycle at G1 phase increasing apoptosis rate. These effects within certain extent are dose and time dependent.5-aza-2-deoxycytidine also coordinates with other chemotherapy drugs in the anti-tumor effects. In nude mouse model the tumor occurrence rate decreased in 5-aza-2-deoxycytidine pro-treated cells of QBC939, and ZdCyd also impedes the growth of transplanted tumor. Conclusion 5-aza-2-deoxycytidine inhibits the growth of QBC939 in vivo and in vitro by inducing cell apoptosis and enhances the antitumor effect of chemotherapy drugs.

Hypermethylation of the promoter region is an important mean for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferase are considered to be the main cause of promoter hypermethylation. In order to further explore the epigenetic mechanism of tumor suppressor gene RASSF1A inactivation, 5-aza-2'-deoxycytidine (5-Aza-CdR), a DNA methyltransferase inhibitor, was used to treat the human biliary tract carcinoma cell line QBC-939 at the concentration of 5 micromol/L for 24 h in this study. After the chemical intervention with 5-Aza-CdR, the methylation status in the promoter region of RASSF1A gene was detected by methylation specific PCR (MS-PCR), and the expression alteration of RASSF1A mRNA and protein were observed by RT-PCR and Western Blot respectively. Following the treatment with 5-Aza-CdR, methylation status in the promoter region of RASSF1A gene was reversed from methylation to unmethylation. A 280 bp DNA band which represented RASS1FA expression at transcriptional level and a 40 kDa (1 kDa=0.9921 ku) protein band which represented RASSF1A expression at protein level were detected by RT-PCR and Western Blot respectively in the experimental group cells and there were no corresponding bands in the control group cells. The experimental results suggest that 5-Aza-CdR can induce demethylation in the promoter region of RASSF1A. It can also reverse epigenetic transcriptional silencing caused by DNA methylation and induce the re-expression of RASSF1A in QBC-939. This study also suggest that the mechanism of RASSF1A inactivation is very closely related to the methylation of the promoter region, which may provide a new epigenetic understanding for tumor related gene inactivation and the pathogenesis of biliary tract carcinoma.

Anterior approach refers to a method of hepatectomy which is first to resect the hepatic parenchyma and then to free the liver; hanging maneuver refers to placing a tape before the inferior vena cava for hanging the liver during hepatectomy.In October 2011,anatomical trisectionectomy was performed on a 54-year-old male patient with large hepatocellular carcinoma in the left medical lobe and right lobe with anterior approach and hanging maneuver.The diameter of the tumor was 16 cm,and was in the ⅢA/T3NOM0 stage.The indocyanine green retention at fifteen minutes was 5.4％,and the ratio of hepatic left lateral lobe volume over the standard total liver volume was 44％.The left bile duct was slightly dilated because of the compress of the tumor.The operation started with the isolation and dissection of the inflow vessels,including the right hepatic artery,the right portal vein,the middle hepatic artery,the portal vein branches of left internal lobe.The hepatic parenchyma transection was performed along the fight side of the falciform ligament.A tape was passed between the anterior surface of inferior vena cava and liver,and the liver was suspended during the transection.The left bile duct was cut at the right side of round ligament,and then the middle hepatic vein and the right hepatic vein were resected.The ligaments around the liver were dissected and the right hepatic lobe was removed.Finally,the end-toend anastomosis between the left hepatic duct and the common hepatic duct was performed.The operation lasted for 4 hours and the intra-operative blood loss was 350 mL.The patient was recovered well.At the end of 4 months after surgery,magnetic resonance cholangiopancreatography showed that the anastomosis of the bile duct was unobstructed,and there was no recurrence of tumor inside the liver.

objective To evaluate the clinical effect of touqing capsule associated with Fengchi on the vertebral artery type of cervical spondylosis(CSA)in the attack stage.Methods A total of 100 patients with CSA, according to the doctor order, were randomly divided into a control group and a treatment group respectively, with 50 patients in each. The control group was treated with acupuncture therapy, the treatment group was additionally treated with Touqingcapsule on the basis of the control group, and both groups were treated for 20 days. The hemodynamics was detected by transcranial doppler, the change of TCM syndrome integral was observed before and after the treatment, and the clinical efficacy was evaluated.Results After treatment, the treatment group dizziness (1.10 ± 0.33 vs. 2.01 ± 0.26,t=14.909), neck shoulder ache (0.87 ± 0.16 vs. 1.92 ± 0.69,t=10.266), numbness (0.95 ± 0.14)vs. 1.87 ± 0.16,t=29.844), the total integral (2.16 ± 0.75 vs.3.89 ± 1.14,t=8.756) integral were significantly lower than the control group(P<0.01).Treatment group RVA (40.03 ± 5.89 cm/svs. 34.26 ± 6.26 cm/s,t=4.628), the LVA (42.14 ± 5.42 cm/svs.33.89 ± 6.08 cm/s,t=6.985) rise and PI (0.79 ± 0.14 vs.1.04 ± 0.19,t=7.312) reduction were significantly better than those of the control group (P<0.01). The total effective rate in the treatment group was significantly better than the control group (91.7%(44/48)vs. 74.5%(35/47),χ2=3.863, P=3.863).Conclusion Touqing Capsule associated with Fengchi is effective for CSA. Its functional mechanism is correlated with improvement of haemodynamics of vertebral-basilar artery of the CSA patients.

Objective To study the effect of transfection with antisense DNMT3b gene eukaryotic expression plasmid on the growth of human cholangiocarcinoma cell line QBC939,and to explore the role of DNMT3b in the cholangiocarcinoma tumorigenesis.Methods The constructed antisense DNMT3b gene eukaryotic expression plasmid was transfected into QBC939 cells using liposome.The expression level of DNMT3b protein was detected by Western blot after stable transfection.The growth curves of transfected cells and un-transfected cells were observed by MTT method respectively.The cell proliferation ability was also observed by the test of colony formation in soft agar.The alterations of the cell cycle and the apoptosis rate were detected by FCM.Results Following the transfection,the protein level of DNMT3b decreased significantly;transfection with antisense DNMT3b gene eukaryotic expression plasmid did not affect the cell growth curve of QBC939,and did not decrease the cell colony formation rate(P=0.717);transfection with antisense DNMT3b gene also did not result in cell cycle alterations or induce cell apoptosis(P=0.089).Conclusions Transfection with antisense DNMT3b gene eukaryotic expression plasmid can down-regulate the expression level of DNMT3b in QBC939.It can not affect the growth and proliferation of human cholangiocarcinoma cell line QBC939,nor alter the cell cycle and induce cell apoptosis.

Objective To investigate the minimum local analgesic concentration(MLAC)of epidural hydromorphone combined with bupivacaine for analgesia during the first stage of labor by establishing clinical model.Methods Sixty four labouring parturi-ents at 3-7 cm cervical dilation who requested epidural analgesia were allocated to one of two groups.After a lumbar epidural cath-eter was placed,study participants received 15 mL bupivacaine(n=30),bupivacaine with hydromorphone 200 μg(n=30).The con-centration of bupivacaine was determined by the response of the previous patient using 0 - 100 mm visual analog pain scores, with≤30 mm within 30 min defined as effective.Results Four women were excluded,leaving 30 patients in each of the two groups for analysis.The MLAC of bupivacaine alone was 0.103%(95% CI :0.094%-0.113%).The addition of hydromorphone at doses of 200 μg resulted in significant reduction(P <0.05)in the MLAC of bupivacaine to 0.044%(95% CI :0.034%-0.053%),the difference was significant (P <0.05).There was no significant difference in Bromage scores,HR,BP,FHR,UC and adverse reac-tion (P >0.05).Conclusion The study showed a significant reduction in the MLAC of bupivacaine by hydromorphone.Hydromor-phone could be safely used in epidural analgesia for labor.

Objective To explore the expression and relationship of PTEN and mTOR in the development of cholangiocarcinoma,determine the effect of the PI3K/PTEN/AKT/mTOR signal transduction on the(development) of cholangiocarcinoma.Methods The expression of mTOR and PTEN in the cholangiocarcinoma was detected by the immunohistochemistry and RT-PCR method.Results Compared to normal tissues,the(expression) of mTOR gene in cholangiocarcinoma significantly increased,but the expression of PTEN gene(decreased).There was a negative correlation between mTOR gene and PTEN gene expression in(cholangiocarcinoma). Conclusions The expression of mTOR gene in cholangiocarcinoma was increased and the expression of PTEN was decreased.It suggested that the mTOR gene and PTEN gene could play an important role in the process of development of cholangiocarcinoma.

Objective To study the toxicity and efficacy of induction chemotherapy followed by concurrent cisplatin chemotherapy and three dimensional conformal radiotherapy (3DCRT) for inoperable locally advanced non-small cell lung cancer (LA-NSCLC). Methods Totally 76 patients with LA-NSCLC received two cycles induction chemotherapy followed by 3DCRT with a median dose of 68 Gy (64 to 74 Gy).During the 3DCRT, cisplatin (25 mg/m2, weekly) was given intravenously for 6-7 times. Results The CR rate, PR rate and overall response rate of induction chemotherapy were 3% ,42% and 45%. After the concurrent chemoradiation, the corresponding figures were 10%, 62% and 72%. The median survival time (MST) and median progression-free survival (PFS) of all patients were 16.6 months and 10.3 months. The 1-, 2- and 3-year overall survival (OS) and PFS rates were 67% , 35% , 21% and 42% , 15%, 6%. Of patients with stage ⅢA and stage ⅢB disease,the MST were 19.7 months and 15.6 months, the PFS were 10.8 months and 9.4 months. The major treatment-related toxicities included radiation esophagitis, radiation pneumonitis, nausea ( or vomiting) and leukopenia. The major pattern of treatment failure was distant metastasis. Forty-five patients (59%) experienced the local recurrence or/and distant metastasis, including 4 (9%) with in-field failure, 38 (84%) distant metastasis and 3 (7%) malignant pleural effusion. Conclusions Induction chemotherapy followed by concurrent weekly cisplatin and 3DCRT for inoperable locally advanced NSCLC results in encouraging outcomes and acceptable tolerance.

Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver cancer with a very poor prognosis.In order to guide better clinical management of ICC patients,the American Joint Committee on Cancer (AJCC) cancer staging manual (7th edition) have established a unique TNM staging scheme for separating ICC from hepatocellular carcinoma (HCC) for the first time,and reflected a difference between risk factor of ICC and HCC.This TNM staging system for ICC has been most recently updated by the AJCC cancer staging manual (8th edition),in which T staging has been redefined without gross features,and lymph node metastasis (N1) in N staging has been grouped as stage Ⅲ B,but not stage Ⅳ as required by the 7th edition of AJCC cancer staging manual.In addition,region lymphatic and distant metastases have been clearly redefined by the AJCC cancer staging manual (8th edition) that also requires recovering at least 6 lymph nodes for the N staging scheme.The apparent advantages of the AJCC cancer staging manual (8th edition) for ICC pathologic staging may better stratify the prognosis of ICC patients and provide an improved guidance in clinical practice.

Hypermethylation of the promoter region is an important mean for the transcriptional repression of a number of cancer-associated genes, and over-expression and/or increased activity of DNA methyltransferase are considered to be the main cause of promoter hypermethylation. In order to further explore the epigenetic mechanism of tumor suppressor gene RASSF1A inactivation,5-aza-2'-deoxycytidine (5-Aza-CdR), a DNA methyltransferase inhibitor, was used to treat the human biliary tract carcinoma cell line QBC-939 at the concentration of 5 μmol/L for 24 h in this study. After the chemical intervention with 5-Aza-CdR, the methylation status in the promoter region of RASSF1A gene was detected by methylation specific PCR (MS-PCR), and the expression alteration of RASSF1A mRNA and protein were observed by RT-PCR and Western Blot respectively. Following the treatment with 5-Aza-CdR, methylaiton status in the promoter region of RASSF1A gene was reversed from methylation to unmethylation. A 280 bp DNA band which represented RASS1FA expression at transcriptional level and a 40 kDa (1kDa=0.9921 ku) protein band which represented RASSF1A expression at protein level were detected by RT-PCR and Western Blot respectively in the experimental group cells and there were no corresponding bands in the control group cells. The experimental results suggest that 5-Aza-CdR can induce demethylation in the promoter region of RASSF1A. It can also reverse epigenetic transcriptional silencing caused by DNA methylation and induce the re-expression of RASSF1A in QBC-939. This study also suggest that the mechanism of RASSF1A inactivation is very closely related to the methylation of the promoter region, which may provide a new epigenetic understanding for tumor related gene inactivation and the pathogenesis of biliary tract carcinoma.