Objective To investigate and analyze the expression of miR-21 in hepatocarcinoma cell strains and the relationship with PTEN. Methods The quantitative reverse transcription-PCR were used to detect the expression of miR-122 and PTEN in hepatocarcinoma cell strains， PTEN and p-Akt expression was detected in tumor-bearing nude mice treated with miR-21 inhibitor. Results The relative expression of miR-21 was 2.97±0.79，137.66±23.60 and 511.21±1.14 in WRL-68， MHCC97-H and SMMC-7721 strains respectively， while that was 1.29±0.33， 36.90±11.84 and 6.22±0.85 respectively when miR-21 inhibitor was transfected. The relative expression of PTEN was 2.88±0.93， 0.19±0.07 and 0.62±0.06 in WRL-68， MHCC97-H and SMMC-7721 strains respectively， while that was 6.11±1.07， 2.65±0.66 and 4.32±0.84 respectively when miR-21 inhibitor was transfected. The expression of PTEN was positive and p-Akt was down regulated when tumor-bearing nude mice was treated with miR-21 inhibitor. Conclusion MiR-21 is an upstream regulation factor of PTEN， it can affect PTEN expression.

Objective To explore the influence on function and activity of rituximab of Raji cells via 4-1BBL gene transfection. Methods Stable expression of 4-1BBL in Raji cells was carried out through transfection mediated by liposome. The experiments contained 4-1BBL group，Mock group and control group. The 4-1BBL expression was identified by Western blotting analysis at 48h after transfection. CCK-8 method was used to detect the proliferation at 48h after treatment with different concentrations （20.0， 10.0， 5.0， 2.5， 1.25， 0.625 mg／ml） of rituximab. The proliferation rates of lymphocyte and supernatant IL-2 levels at 48h after co-culture with the activated peripheral blood lymphocytes under the effector／target ratio of 5∶1 and 10∶1 by CCK-8 and enzyme-linked immunosorbent assay. Results The protein level of 4-1BBL in 4-1BBL group was higher than those in Mock group and control group （P＜0.05）. The proliferative rates decreased with the increasing concentrations of rituximab in 4-1BBL group. When the concentration was over 2.5mg／ml， there were lower proliferative rate in 4-1BBL group versus Mock group and control group （P＜0.05）. Under the effector／target ratio of 5∶1 and 10∶1， the proliferative rate of lymphocyte and supernatant IL-2 level in 4-1BBL group was higher than those in Mock group and control group （P＜0.05）. Conclusion The 4-1BBL gene transfection can enhance the activity of rituximab on Raji cells， suggesting that combined activation of 4-1BBL and targeting-antibody may be a new strategy for cancer immunotherapy.

Objective To investigate the autologous cytokine-induced killer（CIK） cells on the immunity function of cancer patients in different stages. Methods One hundred and sixty-two malignant tumor patients were divided into 3 groups， terminalstage patients and the expected survival time less than 3 months were the group A， the patients who existed distant metastases or locally advanced cancer and the expected survival time longer than 3 months were the group B， and the cases who finished postoperative adjuvantherapy were the group C， meanwhile the health volunteers were as the group D. We detected immunity function of the patients，including the levels of CD3+, CD3+CD4+, CD3+CD8+, CD3+CD56+T cells and the ratio of CD4+／CD8+，meanwhile the adverse reactions were monitored. Results Before the CIK cells therapy， the baseline level of CD3+， CD3+CD4+，CD3+CD8+T cell subsets and the ratio of CD4+／CD8+ among the groups were statistically significant differences（P＜0.05）. After twice CIK cells therapy， the level of CD3+T cells decreased in group A（P＜0.05）; and compared with group D， the levels of CD3+， CD3+CD4+， CD3+CD8+， CD3+CD56+ and CD4+／CD8+were still significantly different（P＜0.05）. The levels of CD3+， CD3+CD56+T lymphocytes increased（P＜0.05） in group B， CD3+，CD3+CD4+ and CD3+CD8+ were still significantly different（P＜0.05）. The levels of CD3+， CD3+CD4+， CD3+CD56+T cell subsets also increased（P＜0.05）in group C， and CD3+， CD3+CD4+， CD3+CD8+， CD3+CD56+ and CD4+／CD8+ were not significantly different compared with group D. The adverse reaction among each groups were in a low rate during treatment and had no statistical significance. Conclusion The autologous CIK cells treatment may help to improve T-lymphocyte subsets distribution and improve the host's immune functions. Meanwhile the adverse reactions in the process of treatment can be alleviated by heteropathy and has no relationship with the status of tumor.

Objective To evaluate the efficacy and safety of endoscopic

resection for submucosal tumors（SMT） of upper gastrointestinal tract and to

analyze histological type of SMT. Methods Of 426 SMTs， 346 located in esophagus， 79 in stomach， 1 in duodenum. By endoscopic ultrasonography（EUS）， 284 lesions were in muscularis mucosa，87 in submucosal layer， 55 in muscularis propia. Lesions ranged from 0.4cm to 6cm（0.90±0.03cm）. Three hundred and thirtythree patients were managed by endoscopic mucosal resection（EMR）， 37 by endoscopic submucosal dissection（ESD）， 47 by endoscopic submucosal excavation（ESE）， and 9 by endoscopic fullthickness resection（EFR）. Results In EMR group， 314 out of 333 lesions（94.3％） were en bloc resected， compared with 100.0％ in ESD group and EFR group，and 44 out of 47 lesions（93.6％） in ESE group. Three cases（2 complicated with unmanaged perforation，1 with maximum diameter reaching 6cm） from ESE group performed surgery for unsuccessful endoscopic resection. 1 case from ESD and ESE group each with delayed hemorrhage had endoscopic hemostatic therapy without surgery. Nine active perforation in EFR group were all successfully managed by endoscopy. Three hundred and twenty-six lesions were histopathologically diagnosed as leiomyoma， 58 as gastrointestinal stromal tumors. No case relapsed in 2 years followed-up. Conclusion Endoscopic resection can be a safe and effective treatment for upper gastrointestinal tract SMT.

Objective To investigate the expression of 6-phosphofructo-2kinase／fructose-2，6-bisphosphatase （PFKFB3） in hepatocellular carcinoma（HCC） and its clinical significance. Methods The expressions of PFKFB3 mRNA of tissue sections from 42 HCC and paired adjacent tissues were detected by RT-PCR. Then the relationships between PFKFB3 expression and clinicopathological variables （gender， age， tumor size， lymphatic metastasis， cancer embolus in vein， AFP level before treatment， differentiation and metastasis in liver） were analyzed. Meanwhile， the difference of overall survival of patients with different expression of PFKFB3 were studied by survival analysis of Kaplanmeier. Results The positive expression rate and relative level of PFKFB3 in HCC tissue were 73.81％ （31／42） and 1.075±0.171， higher than 52.38％ （22／42） and 0.877±0.451 in adjacent tissues with significant difference （P＜0.05）. The PFKFB3 expression is related with TNM stage， degree of ifferentiation， intrahepatic metastasis and overall survival （P＜0.05）. Conclusion PFKFB3 were related to carcinogenesis， development and metastasis of HCC. The expression of PFKFB3 can influence the differentiation， metastasis andprognosis of HCC， indicating it may be used as biological indicators to evaluate the degree of malignancy of HCC.