活细胞治疗剂:下一代药物

数家新、老制药公司正在发展一种新疗法,这种新疗法应用经过体外处理的活细胞,而不是传统的药物.虽然“活细胞疗法”包括几种不同的技术,但是这些技术加在一起能治疗许多疾病,这些疾病每年折磨着全世界一百多万人。新疗法治疗的疾病包括癌症、遗传性疾病、糖尿病、血友病、烫伤和艾滋病。活细胞的处理有多种多样,从简单地增殖患者自身细胞到遗传工程细胞产生治疗性蛋白质,如胰岛素。目前活细胞疗法包括六种基本方法:体外淋巴细胞处理法;骨髓细胞增殖;培养细胞产生的人造组织;基因疗法;胎细胞移植和细胞包裹法。当前,仅有少数公司活跃在上述任何一个领域内。

Effects of Low Dose Radiation on Tumor Apoptosis, Cell Cycle and Apoptosis-Related Protein Bcl-2 in Tumor-Bearing Mice

To study the effects of low dose radiation (LDR) on tumor apoptosis, cellcycle progression and changes of apoptosis-related protein Bcl-2 in tumor-bearing mice. Methods:Male mice of Kunming strain were implanted subcutaneously with S180 sarcoma cells in the left inguenas an in situ experimental animal model. Seven days later, the mice were subjected to 75 mGywhole-body γ-irradiation. At 24 and 48 h after the irradiation, all mice were sacrificed. The tumorsizes were measured, and tumor cell apoptosis and cell cycle progression were analyzed by flowcytometry. The expression of apoptosis-related protein Bcl-2 and the apoptotic rate of tumor cellswere observed by immunohistochemistry and electron microscopy. Results: Tumors grew significantlyslower after LDR (P 【 0.05). The tumor cells were arrested in G1 phrase and the expression of Bcl-2protein decreased at 24 h. Apoptotic rate of tumor cells was increased significantly at 48 h afterLDR (P 【 0.01). Conclusion: LDR could cause a G1-phase arrest and increase the apoptosis of tumorcells through the low level of apoptosis-related protein bcl-2 in the tumor-bearing mice. Theorganized immune function and anti-tumor ability are markedly increased after LDR. Our studyprovides practical evidence of clinical application to cancer treatment.

Molecular mechanism underlying the functional loss of cyclindependent kinase inhibitors p16 and p27 in hepatocellular carcinoma

Hepatocellular carcinoma （HCC） is one of the most common human cancers, and its incidence is still increasing in many countries. The prognosis of HCC patients remains poor, and identification of useful molecular prognostic markers is required. Many recent studies have shown that functional alterations of cellcycle regulators can be observed in HCC. Among the various types of cell-cycle regulators, p16 and p27 are frequently inactivated in HCC and are considered to be potent tumor suppressors, p16, a G1-specific cell-cycle inhibitor that prevents the association of cyclindependent kinase （CDK） 4 and CDK6 with cyclin DI, is frequently inactivated in HCC via CpG methylation of its promoter region, p16 may be involved in the early steps of hepatocarcinogenesis, since p16 gene methylation has been detected in subsets of pre-neoplastic liver cirrhosis patients, p27, a negative regulator of the G1-S phase transition through inhibition of the kinase activities of Cdk2/cyclin A and Cdk2/cyclin E complexes, is now considered to be an adverse prognostic factor in HCC. In some cases of HCC with increased cell proliferation, p27 is overexpressed but inactivated by sequestration into cyclin D1-CDK4-containing complexes. Since loss of p16 is closely related to functional inactivation of p27 in HCC, investigating both p16 and p27 may be useful for precise prognostic predictions in individuals with HCC.

Impact of prolonged fraction dose-delivery time modeling intensity-modulated radiation therapy on hepatocellular carcinoma cell killing

AIM: To explore the impact of prolonged fraction dosedelivery time modeling intensity-modulated radiation therapy (IMRT) on cell killing of human hepatocellular carcinoma (HCC) HepG2 and Hep3B cell lines.METHODS: The radiobiological characteristics of human HCC HepG2 and Hep3b cell lines were studied with standard clonogenic assays, using standard linear-quadratic model and incomplete repair model to fit the dose-survival curves. The identical methods were also employed to investigate the biological effectiveness of irradiation protocols modeling clinical conventional fractionated external beam radiotherapy (EBRT, fraction delivery time 3 min) and IMRT with different prolonged fraction delivery time (15, 30, and 45 min). The differences of cell surviving fraction irradiated with different fraction delivery time were tested with paired t-test. Factors determining the impact of prolonged fraction delivery time on cell killing were analyzed.RESULTS: The α/β and repair half-time (T1/2) of HepG2and Hep3b were 3.1 and 7.4 Gy, and 22 and 19 min respectively. The surviving fraction of HepG2 irradiated modeling IMRT with different fraction delivery time was significantly higher than irradiated modeling EBRT and the cell survival increased more pronouncedly with the fraction delivery time prolonged from 15 to 45 min,while no significant differences of cell survival in Hep3b were found between different fraction delivery time protocols.CONCLUSION: The prolonged fraction delivery time modeling IMRT significantly decreased the cell killing in HepG2 but not in Hep3b. The capability of sub-lethal damage repair was the predominant factor determining the cell killing decrease. These effects, if confirmed by clinical studies, should be considered in designing IMRT treatments for HCC.

Rare pulmonary and cerebral complications after transarterial chemoembolisation for hepatocellular carcinoma:A case report

We report a rare case of acute pulmonary and cerebral complication after transarterial chemoembolisation (TACE) for inoperable hepatocellular carcinoma. The case involved a large tumor and hepatic vein invasion. Nonspecific pulmonary and cerebral symptoms such as acute dyspnoea and transient consciousness loss developed in the patient, a 49-year-old woman, following the TACE due to pulmonary and cerebral oil embolism. The chest and brain conditions of this patient improved after some supportive therapies and nursing interventions. She also subsequently completed the other three procedures of TACE.

Risk factors for immediate post-operative fatal recurrence after curative resection of hepatocellular carcinoma

AIM： To investigate the clinicopathological risk factors for immediate post-operative fatal recurrence of hepatocellular carcinoma （HCC）, which may have practical implication and contribute to establishing high risk patients for pre- or post-operative preventive measures against HCC recurrence. METHODS： From June 1994 to May 2004, 269 patients who received curative resection for HCC were reviewed. Of these patients, those who demonstrated diffuse intrahepatic or multiple systemic recurrent lesions within 6 mo after surgery were investigated （fatal recurrence group）. The remaining patients were designated as the control group, and the two groups were compared for clinicopathologic risk factors. RESULTS： Among the 269 patients reviewed, 30 patients were enrolled in the fatal recurrence group. Among the latter, 20 patients showed diffuse intrahepatic recurrence type and 10 showed multiple systemic recurrence type. Multivariate analysis between the fatal recurrence group and control group showed that preoperative serum alpha-fetoprotein （AFP） level was greater than 1 000 μg/L （P= 0.02; odds ratio = 2.98）, tumor size greater than 6.5 cm （P= 0.03; OR= 2.98）, and presence of microvascular invasion （P= 0.01; OR=4.89） were the risk factors in the fatal recurrence group. The 48.1% of the patients who had all the three risk factors and the 220 of those who had two risk factors experienced fatal recurrence within 6 mo after surgery. CONCLUSION： Three distinct risk factors for immediate post-operative fatal recurrence of HCC after curative resection are pre-operative serum AFP level 〉 1 000 μg/L,tumor size〉6.5 cm, and microvascular invasion. The high risk patients with two or more risk factors should be the candidates for various adjuvant clinical trials.

Tumor suppressor and hepatocellular carcinoma

A few signaling pathways are driving the growth of hepatocellular carcinoma.Each of these pathways possesses negative regulators.These enzymes,which normally suppress unchecked cell proliferation,are circumvented in the oncogenic process,either the overactivity of oncogenes is sufficient to annihilate the activity of tumor suppressors or tumor suppressors have been rendered ineffective.The loss of several key tumor suppressors has been described in hepatocellular carcinoma.Here,we systematically review the evidence implicating tumor suppressors in the development of hepatocellular carcinoma.

Diagnosis and therapy of ascites in liver cirrhosis

Ascites is one of the major complications of liver cirrhosis and is associated with a poor prognosis. It is important to distinguish noncirrhotic from cirrhotic causes of ascites to guide therapy in patients with noncirrhotic ascites. Mild to moderate ascites is treated by salt restriction and diuretic therapy. The diuretic of choice is spironolactone. A combination treatment with furosemide might be necessary in patients who do not respond to spironolactone alone. Tense ascites is treated by paracentesis, followed by albumin infusion and diuretic therapy. Treatment options for refractory ascites include repeated paracentesis and transjugular intrahepatic portosystemic shunt placement in patients with a preserved liver function. Potential complications of ascites are spontaneous bacterial peritonitis (SBP) and hepatorenal syndrome (HRS). SBP is diagnosed by an ascitic neutrophil count > 250 cells/mm3 and is treated with antibiotics. Patients who survive a first episode of SBP or with a low protein concentration in the ascitic fluid require an antibiotic prophylaxis. The prognosis of untreated HRS type 1 is grave. Treatment consists of a combination of terlipressin and albumin. Hemodialysis might serve in selected patients as a bridging therapy to liver transplantation. Liver transplantation should be considered in all patients with ascites and liver cirrhosis.

Effect of Yiguanjian decoction on cell differentiation and proliferation in CCl_4-treated mice

AIM： To investigate the cellular mechanisms of action of Yiguanjian （YGJ） decoction in treatment of chronic hepatic injury. METHODS： One group of mice was irradiated, and received enhanced green fluorescent protein （EGFP）- positive bone marrow transplants followed by 13 wk of CCh injection and 6 wk of oral YGJ administration. A second group of Institute for Cancer Research mice was treated with 13 wk of CCI4 injection and 6 wk of oral YGJadministration. Liver function, histological changes in the liver, and Hyp content were analyzed. The expres- sion of m-smooth muscle actin （α-SMA）, F4/80, albumin （AIb）, EGFP, mitogen-activated protein kinase-2 （PKM2）, Ki-67, fetoprotein （AFP）, monocyte chemotaxis pro- tein-1 and CC chemokine receptor 2 were assayed. RESULTS： As hepatic damage progressed, EGFP-po- sitive marrow cells migrated into the liver and were mainly distributed along the fibrous septa. They showed a conspicuous coexpression of EGFP with ^-SMA and F4/80 but no coexpression with AIb. Moreover, the expression of PKM2, AFP and Ki-67 was enhanced dy- namically and steadily over the course of liver injury. YGJ abrogated the increases in the number of bone marrow-derived fibrogenic cells in the liver, inhibited expression of both progenitor and mature hepatocyte markers, and reduced fibrogenesis. CONCLUSION： YGJ decoction improves liver fibrosis by inhibiting the migration of bone marrow cells into the liver as well as inhibiting their differentiation and suppressing the proliferation of both progenitors and hepatocytes in the injured liver.

Comparative study of IORT and 3D-CRT for hepato-cellular carcinoma

Objective: To investigate the feasibility of intra-operative radiation therapy (IORT) in hepato-cellular carcinoma (HCC). Methods: Based on the dosage distribution of tumor and adjacent sensitive organs, and dose-volume histogram (DVH), Topslane three-dimensional therapy plan was used to compare IORT and three-dimensional conformal radiation therapy (3D-CRT) in 12 cases of HCC. Results: Taking the center of tumors as the isodose center, the V90 (volume of 90% dose distribution) of IORT was significantly better than that of 3D-CRT, and median absorbed doses of normal organs in IORT was significantly lower than that in 3D-CRT. Conclusion: The V90 of IORT is better than that of 3D-CRT in HCC, and neighboring sensitive organs were effectively protected by IORT. The tumors absorbed dose and local control rate are improved in IORT.

The effects of low-dose splenic irradiation and radiotherapy on immune system of patients with locally advanced non-small cell lung cancer

Objective： The aim of the research was to study the effects of low-dose splenic irradiation and radiotherapy on immune system of patients with locally advanced non-small cell lung cancer （NSCLC）. Methods： Twelve cases of stage III NSCLC in Tumor Radiotherapy Center of our hospital （the Affiliated Hospital of Medical College Qingdao University, China） were collected from July 2011 to July 2012; all patients were under 75 years old with clear pathology, measurable lesions and good personal statement. They were randomly divided into combined treatment group （D1 ＋ D2） and control group （D1）. The control group （D1） only received radiotherapy to the chest; combined treatment group （D1 ＋ D2） received low-dose splenic irradiation plus conventional dose irradiation. Flow cytometry was used to detect the peripheral blood T lymphocyte immune indexes of patients before, during and after the treatment, classification by five blood cell analyzer was used to determine white blood cells, neutrophils, hemoglobin and platelet count. The radiation induced toxicity including esophagitis, pneumonia and gastrointestinal reaction was observed, as well as the dose when it happened. Results： There was no significant difference in the ratio between two groups in cells CD4＋, CD8＋ and CD4＋/CD8＋ after radiotherapy （P 〉 0.05）. There was no change in these indicators in combined treatment group after treatment （P 〉 0.05）, but it decreased in control group （P 〈 0.05）. There was no significant difference in the incidences of radiation esophagitis, pneumonia, gastrointestinal reactions and bone marrow suppression between two groups （P 〉 0.05）, but the patients in combined treatment group seemed to tolerate high dose well （P 〈 0.05）. Conclusion： Low-dose splenic irradiation combined with radiotherapy to the chest can alleviate the injury degree of acute radiation induced the toxicity of locally advanced NSCLC patients, through affect the patient＇s immune function.

Observation of Intensity-Modulated Radiotherapy with Different Fractionated Doses Used in 58 Cases with Astrocytoma

OBJECTIVE To analyze the therapeutic effects and side effects of intensity-modulated radiotherapy （IMRT） with different fractionated doses in treating astrocytoma.METHODS During a period from October 2001 to December 2006, 58 patients with astrocytoma were treated using IMRT. Based on the World Health Organization （WHO） classification, 32 of the 58 cases were grade-Ⅱ, 20 grade-Ⅲ and 6 grade-IV （glioblastoma multiforme, GBM）. Thirty-two of the 58 patients （3 with grade IV, 11 with grade Ⅲ, and the other 18 with grade II who were over 40 years） were treated with hyperfractionated IMRT （Hyper Fr IMRT）, and the other 26 patients were treated with standard fractionated IMRT （St Fr IMRT）.RESULTS The 1-, 3- and 5-year overall survival （OS） rates were respectively 86%, 52%, and 45%, and the 1-, 3- and 5-year progression-free survival （PFS） rates were respectively 77%, 38%, and 25%. Using an analytical hierarchy process it was shown that concerning the patients with grade II astrocytoma classified based on WHO grading, the therapeutic effect was much better in the group of Hyper Fr IMRT than in the St Fr IMRT group. There was no statistical significance of the differences in the OS and PFS rates between the 2 groups （P = 0.049 and P = 0.006）. The OS and PFS rates of the patients with grade-III astrocytoma were both higher in the group with Hyper Fr IMRT than in the St Fr IMRT group. However, there was no statistical significance of the differences between the 2 groups. Advanced RTOG grade-Ⅲ（radiation therapy oncology group, RTOG） neurotoxicity occurred only in 1 of the cases.CONCLUSION Compared with the St Fr IMRT, the Hyper Fr IMRT may help to prolong the survival of patients with astrocytoma.

Molecular targeted therapy in gastrointestinal cancer

Gastrointestinal cancer is one of the highly prevalent malignant diseases worldwide which is a major cause of morbidity and mortality. Gastric cancer is the second leading cause of cancer mortality in the world and its management, especially in advanced stages, has evolved relatively little [1]. Colorectal cancer (CRC) remains the third most common ma-lignancy and the third leading cause of cancer death worldwide [2]. The surgical treatment is still the most effective therapy for the gastrointestinal cancer. However, the majority of the patients had lost the opporunity of surgical therapy when it was detected at advanced stage, so to seek means other than surgical treatment of gastrointestinal cancer metastasis and recur-rence also has an important significance. With the deeping research of the molecular biology, molecular targeted therapy has become the hotspot and focus of comprehensive treatment of gastrointestinal cancer which is proposed against the molecular biological targets such as tumor cell growth, apoptosis, cell cycle, invasion and angiogenesis. Molecular targeted therapy can be grouped into six main areas: the epidermal growth factor receptor (EGFR) inhibitors, anti-angiogenic factors, cell cycle inhibitors, apoptosis promoters and matrix metalloproteinase inhibitors, cyclooxygenase inhibitors. The review of the progress are as follows.

Research progress in immunotherapy of pancreatic cancer

Pancreatic cancer is among the most lethal malignancies resistant to conventional therapies. The urgent need fornew therapies has turned the spotlights on immunotherapy. In recent years, a growing body of evidence has alreadybeen gathered regarding the efficacy of genetic engineering modified T-cells, checkpoint inhibitors of T-cells, killercells induced by dendritic cells and cytokine in patients with pancreatic cancer. Cryoimmunotherapy in situ andextra-tumor and immunotherapy combined with chemotherapy could also increase the effectiveness. Research ofpancreatic cancer vaccine has made some progress. The immunity enhancing function of some traditional herbshave been reported, such as Ginsenoside Rg3, which could enhance T-cell subsets and NK cell activity inpancreatic cancer patients with chemotherapy.

Anti-inflammatory effect of erythropoietin pretreatment on cardiomyocytes with hypoxia/reoxygenation injury and the possible mechanism

Objective： To investigate the anti-inflammatory effect of erythropoietin （EPO） pretreatment on cardiomyocytes exposed to hypoxialreoxygenation injury （H/R） and explore the possible mechanism. Methods： The cultured neonatal rats＇ ventricular cardiomyocytes were divided randomly into 4 groups, control group （C group）, EPO pretreatment group （E group）, EPO and pyrrolidine dithiocarbamate （PDTC） pretreatment group （EP group） and PDTC pretreatment group （P group）. After 24 hours＇ pretreatment, the cardiomyocytes were exposed to H/R. After pretreatment and H/R, the expression of tumor necrosis factor- α （TNF- α ） gene in all the groups was detected by RT-PCR and Western blot. The nuclear factor- κ B （NF- κB） activity was detected by electrophoretic mobility shift assay （EMSA） and the inhibitor- κB α （Ⅰ- κB α） protein level was detected by Western blot. Results： The decrement of Ⅰ- κB a protein and the increasing NF- KB activity were found in cardiomyocytes pretreated with EPO before H/R compared to other groups （t=3.321, 4.183, P〈0.01）. However, after H/R, NF- κB activity and expression of TNF- α gene were significantly reduced, Ⅰ- κB a protein expression was increased in cardiomyocytes of E group compared to other groups （t=-3.425, 3.687, 3.454, P〈0.01）. All theses changes caused by EPO pretreatment were eliminated by the intervention of PDTC （an antagonist to NF- κB） during pretreatment. Conclusions： EPO pretreatment can inhibit the activation of NF- κB and upregulation of TNF- α gene in cardiomyocytes exposed to H/R through a negative feedback of NF- κB signaling pathway, and thus produces the anti-inflammatory effect. This might be one of the ways EPO produces the anti-inflammatory effect.

Mechanism research on combination of decoction for reinforcing lung Qi and argon helium lancet in treatment of non-small cell lung cancer

OBJECTIVE: To observe the effect of decoction for reinforcing lung Qi on T-lymphocytic function, interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α) in peripheral blood of patients with non-small cell lung cancer after operation with argon helium lancet in order to explore its mechanism.METHODS: A total of 76 patients suffering from non-small cell lung cancer without surgical indication were randomly divided into a treatment group treated with decoction for reinforcing lung Qi and argon helium lancet and a control group treated with argon helium lancet only to observe lymphocytic proliferation, detect the percentage of positive cells in the T-lymphocyte CD28 with flow cytometry and detect the expression of IL-2 and TNF-α in peripheral blood with enzyme linked immunosorbent assay.RESULTS: Proliferation of T-lymphocytes and expression of CD28, IL-2 and TNF-α in peripheral blood after treatment in the treatment group were more obviously strengthened than those before treatment and those in the control group (all P<0.05).CONCLUSION: The mechanism of using decoction for reinforcing lung Qi and argon helium lancet to treat lung cancer may be realized through promoting T-lymphocytic proliferation, up-regulating expression of CD28, IL-2 and TNF-α, and activating T-cells.