1.Expression and activities analysis of a fusion protein CREKA/tTF
Yi SU ; Jianghua YAN ; Shengyu WANG ; Jie HE ; Min YE
Chinese Journal of Biochemical Pharmaceutics 2010;31(2):94-97
Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.
3.Effect of Helicobacter pylori on the function of peripheral blood monocyte-derived dendritic cells in gastric cancer patients
Qiaozhen LI ; Yuqiang CHEN ; Jianghua YAN ; Yuan DING ; Yongjun WANG ; Shengyu WANG ; Yanhua YE ; Yingyi XU ; Huixiang HE
Chinese Journal of Clinical Oncology 2013;(15):902-906
Objective: This study aimed to compare and analyze the functional differences between peripheral blood mono-cyte-derived dendritic cells (DCs) of Helicobacter pylori-positive and H. pylori-negative patients with gastric cancer. Methods:H. py-lori infection was detected in 84 patients with gastric cancer in our hospital from January 2011 to October 2012 by the 14C-urea breath test. DCs were generated from monocytes isolated by an adherent method from the two groups of patients and cultured in the presence of rhIL-4, rhGM-CSF, and rhTNF-α. Furthermore, the expression of surface marker molecules was determined by fluorescence-activat-ed cell sorting analysis. The cytotoxicity of DCs pulsed T cells against gastric carcinoma cell was assessed by the lactate dehydroge-nase-releasing assay. The secretion of IL-12 and IFN-γin the supernatant was determined by enzyme-linked immunosorbent assay. Re-sults:No difference was observed in the morphological change of the maturation process. The mean expression of CD1a, CD80, CD83, CD86, and HLA-DR molecules in DCs of H. pylori-infected patients was higher than that in DCs of H. pylori-negative group, and the differences were statistically significant except for CD1a and HLA-DR. The cytotoxicity activities, IL-12 release, and IFN-γrelease in the H. pylori-positive group were significantly higher than those in the H. pylori-negative group (P<0.05). Conclusion:H. pylori infec-tion has no effect on the morphological change of the maturation process of monocyte-derived DCs. These data clearly demonstrate that monocyte-derived DCs of H. pylori-infected patients with gastric cancer can induce stronger maturation and activation than those of H. pylori-negative patients.
4.Efficacy analysis of pegylated filgrastim as prophylaxis for chemo-therapy-induced neutropenia
Sheng YANG ; Xiaohui HE ; Peng LIU ; Shengyu ZHOU ; Mei DONG ; Yan QIN ; Jianliang YANG ; Changgong ZHANG ; Xiaohong HAN ; Yuankai SHI
Chinese Journal of Clinical Oncology 2015;(12):626-631
Objective:To compare the efficacy of a single injection of pegylated filgrastim with daily doses of filgrastim as pro-phylaxis for chemotherapy-induced neutropenia in Chinese cancer patients. Methods:Single-institution data from a phase 2 study and a phase 3 trial on pegylated filgrastim were combined to analyze the efficacy and safety parameters. In the two randomized crossover tri-als, patients with previously untreated cancers received two cycles of chemotherapy with identical regimen. In the study cycle, the pa-tients received a single subcutaneous injection of pegylated filgrastim (100 μg/kg), whereas those in the control cycle received daily subcutaneous injections of filgrastim (5μg/kg). Results:Among the 56 patients enrolled, 53 were evaluable for efficacy. These patients received one cycle with pegylated filgrastim prophylaxis and one cycle with filgrastim support each. Results indicated that 94.3%(50/53) of the cycles with pegylated filgrastim or filgrastim support did not develop grade 4 neutropenia. Moreover, febrile neutropenia did not occur in the cycles. The incidence rates of antibiotic administration were 7.5%(4/53) and 3.8%(2/53) in the pegylated filgrastim and filgrastim cycles, respectively (P=0.678). The median duration of filgrastim administration was 10 days (3-14 days). Generally, the safety profile of pegylated filgrastim is similar to that of filgrastim, including skeletal pain, pain at the injection site, palpitation, fever, and fatigue. Conclusion:A single dose of pegylated filgrastim demonstrated comparable efficacy with 10 consecutive doses of filgras-tim as prophylaxis for chemotherapy-induced neutropenia.
5.Retrospective analysis of the clinical features and prognostic factors of 126 patients with primary gastric diffuse large B-cell lymphoma
Yan QIN ; Xiaohui HE ; Shengyu ZHOU ; Peng LIU ; Jianliang YANG ; Changgong ZHANG ; Sheng YANG ; Lin GUI ; Yuankai SHI
Chinese Journal of Clinical Oncology 2016;43(14):620-625
Objective:Primary gastric diffuse large B-cell lymphoma (PGLBCL) is a highly common subtype of extranodal non-Hodgkin lymphoma. We analyzed the disease's clinical features and prognosis to guide better treatment. Methods:We retrospectively collect-ed data from PGLBCL cases seen from January 1999 to March 2012 in one cancer center. We then analyzed the demographic character-istics, clinical stage, histological diagnosis, complications, treatment, and prognostic characteristics of such patients. Results:A total of 126 patients with median age of 49 years old (range:16-81 years) were included in the study. The male-to-female ratio was 68:58. A to-tal of 96 patients were pathologically diagnosed with pure diffuse large B-cell lymphoma (DLBCL), 27 with mucosa-assouated lymphoid (MALT) component, and 3 with plasmacytoid differentiation. Meanwhile, 90%of the patients were in the early stage of the disease. For the early-stage patients, treatment strategy included surgery+chemotherapy ± radiotherapy for 38 cases, chemoradiotherapy for 39 cases, chemotherapy alone for 37 cases, and surgery alone for 1 case. Under a median follow up of 48 months, the 4-year progres-sion free survival (PFS) and overall ourvival (OS) rate of the whole group were 75.6%and 82.7%, respectively. PFS rates for early and advanced stage patients were 77%and 41.7%(P=0.005), respectively. For the early-stage patients treated with chemotherapy alone, chemoradiotherapy, and surgery with therapy, the PFS rates were 67.3%, 77.8%, and 77.8%(P=0.588), respectively. The patients with international prognostic index (IPI) score of 0, 1, and>1 achieved PFS of 85.4%, 74.4%, and 55.6%(P=0.011), respectively. The PFS rates were 81.2%and 66.1%(P=0.018) for stagesⅠandⅡ, respectively, and 86.6%and 63.3%(P=0.006) for the normal and elevated LDH levels, respectively. The pathological type of pure DLBCL or a MALT component, GCB or non-GCB origin, and age more than 60 years old were not associated with prognosis. Conclusion:The majority of the PGLBCL patients were in the early stage of disease, but the outcome of early-stage disease was favorable. Surgery did not improve outcomes. Univariate analysis demonstrated that IPI score>1, stageⅡdisease, and elevated LDH levels were associated with poor prognosis in the early-stage patient.
6.Intra-axonal overloading of calcium ion in rat diffuse axonal injury and therapeutic effect of calcium antagonist.
Xiaosheng HE ; Shengyu YI ; Xiang ZHANG ; Zhou FEI ; Jianning ZHANG ; Lisun YANG ; Jingwen LIANG
Chinese Journal of Traumatology 1999;2(1):25-29
OBJECTIVE: Exploring the intra-axonal overloading of calcium ion (Ca(2+)) in brain diffuse axonal injury (DAI) and the therapeutic effect of calcium antagonist(Nimotop) on DAI. METHODS: Fourteen SD rats were divided into injury group, treatment group and control group. The DAI model of rats was produced by using a head-instant-axial-rotation device. Tissues from the medulla oblongata of rats were taken 2-24 h post-injury and processed for electron microscopic observation by a cytochemical technique for calcium ion. RESULTS: In the injured rats there was evidence of local disruption of myelin sheath,lucent spaces between myelin sheath lamellae, separation of axolemma from the inner layer of myelin sheath, peripheral accumulation of organellae, intra-axonal formation of vacuoles and reduction of mitochondria. A large number of fine calcium deposits were seen on the affected myelin sheath. The severity of the myelin sheath lesion was related positively to the number of calcium deposits on it. In the later post-injury period the coarse calcium particles appeared within the damaged axon. Neuronal somas and microvascular endotheliums showed a lot of vacuoles and some fine calcium deposits. Many microvilli formed on the luminal aspect of endothelium. In the treatment group myelin sheath tended to be injured locally, and axoplasmic mitochondria were nearly normal in number, structure, and distribution. Few calcium deposits were found in axons. Vacuolization was obviously reduced in neuronal soma and endothelium. CONCLUSIONS: In DAI there exists an intra-axonal overloading of calcium ion, which is a key factor to the occurrence and development of DAI. Early use of Nimotop can alleviate DAI.
7.Content change of neurofilament protein subunits in experimental brain diffuse axonal injury by lateral head rotation.
Xiaosheng HE ; Shengyu YI ; Xiang ZHANG ; Zhou FEI ; Jianning ZHANG ; Lisun YANG
Chinese Journal of Traumatology 2000;3(1):45-49
OBJECTIVE: To explore the content change of neurofilament (NF) protein subunits in the experimental brain diffuse axonal injury (DAI) by lateral head rotation. METHODS: Twenty-four Sprague Dawley (SD) rats were equally divided into three injury groups (2 h, 12 h, and 24 h post injury) and one control group. The models of DAI were made in the injury groups by lateral head rotation. Western blotting technique was used to measure the content of NF68 (a kind of NF protein subunit) in the brainstem tissues among all the injured and control rats. The NF68 immunohistochemical staining was used in another six SD rats in order to observe the morphological changes in DAI. RESULTS: The NF68 content in the brainstem tended to decrease at 2 h post injury, decreased significantly at 12 h and continued its decrease at 24 h. NF56 and NF52, as the breakdown products of NF68, had a tendency to increase at 2-12 h after the injury, and amounted to a significantly higher level at 24 h. Microscopically, there were a lot of swelling neuronal axons in the ventral part of the medullar oblongata at 2 h after the injury. Some axons were disconnected, and axonal retraction balls formed on their proximal end. CONCLUSIONS: There is an occurrence of phosphorolysis within the brainstem in DAI by lateral head rotation. These reactions cause the breakdown of NF68, which results in the decrease of NF68 in content. It suggests that the breakdown of neurofilament protein subunits is an important reason for structur al destroy of neurofilaments in DAI.
8.Analysis of the serological testing results from Aier Eye Bank
Wenxiu LONG ; Lihong YANG ; Jie XU ; Shengyu HE ; Shaowei LI ; Congxiang WANG ; Kehua WANG ; Ruqiu RAN ; Jian TANG
Chinese Journal of Experimental Ophthalmology 2018;36(5):351-354
Objective This study was to evaluate the safety of 640 corneal donors by analysing the serological testing results.Methods We retrospectively analyzed the serological testing results from Changsha Aier Eye Bank and Chengdu Kangqiao Aier Eye Bank from January 2011 to December 2015,hepatitis B virus surface antigen (HBsAg),hepatitis C virus (HCV),treponema pallidum (TP) and human immunodeficiency virus (HIV) were detected by colloidal gold or enzyme-linked immunosorbent assay (ELISA).Results There were 83 out of 640 serum samples showed positive immuno-reaction assayed markers,the positive rate was 12.97%,including HBsAg(n=60,9.38%),HCV(n=3,0.47%),TP(n=11,1.72%) and HIV(n=2,0.31%).Moreover,3 corneal donors were both positive against HBsAg and HCV,2 donors positive against HCV and TP,1 donor positive against HBsAg and HIV,1 donor positive against HBsAg and TP.Conclusions There is a high proportion of positive results of blood-borne diseases in cornea donors,which is a potential threat to corneal receptors and eye bank workers.Therefore,it is very important to detect serological test strictly for corneal donors.
9.Prolonging use of tranexamic acid is helpful to reduce perioperative hidden blood loss in senile patients with intertrochanteric fractures
Yan ZHAO ; Fan WU ; Hong LI ; Shengyu WAN ; Jin HE ; Binren ZHU ; Congbing JIANG
Chinese Journal of Tissue Engineering Research 2024;28(36):5858-5864
BACKGROUND:Intertrochanteric fracture of femur often occurs in the elderly,and there will be a large amount of hidden blood loss after surgery.Reducing hidden blood loss can decrease complications and hospital stay. OBJECTIVE:To evaluate the effect of prolonged use of tranexamic acid on hidden blood loss after proximal femoral nail antirotation implantation in senile intertrochanteric fractures. METHODS:From January 2022 to May 2023,62 elderly admitted patients with intertrochanteric fracture of femur were selected from Zigong Fourth People's Hospital.All of them were treated with proximal femoral nail antirotation implantation after closed reduction on the traction bed.According to the use time of tranexamic acid,they were divided into two groups.In the control group(n=38),1 g tranexamic acid was given intravenically 15-30 minutes before incision,and 1 g was added 3 hours later.Based on the control group,the trial group(n=24)was given 1 g tranexamic acid intravenously once for 12 hours on the first day after surgery.Blood routine examinations were performed before surgery,on the day after surgery,and on the first,third and fifth days after surgery.Hemoglobin and hematocrit were counted.The theoretical total blood loss was calculated by Cross equation,and the incidence of complications in the two groups was recorded. RESULTS AND CONCLUSION:(1)Through statistical analysis,there was no significant difference in the amount of dominant blood loss between the two groups(P>0.05).(2)The number of grams of hemoglobin decreased,total blood loss and hidden blood loss in the trial group during perioperative period were lower than those in the control group,and the differences were statistically significant(P<0.05).(3)The hemoglobin values of the trial group on day 3 after surgery,and the hematocrit values on days 1 and 3 after surgery were higher than those of the control group,with statistical significance(P<0.05).(4)The hemoglobin and platelet count showed a downward trend after surgery,and the hemoglobin value was the lowest value on day 3,and the platelet value was the lowest value on day 1 after surgery,and then began to rise in both groups.(5)There was no significant difference in postoperative complications between the two groups(P>0.05).(6)The results show that prolonging use of tranatemic acid can effectively reduce the hidden blood loss in the treatment of femoral intertrochanteric fracture with proximal anti-rotation intramedullary nail,and does not increase the risk of complications.
10.Clinical features and prognostic analysis of mantle cell lymphoma patients.
Ping ZHOU ; Yuankai SHI ; Xiaohui HE ; Shengyu ZHOU ; Peng LIU ; Mei DONG ; Yan QIN ; Jianliang YANG ; Changgong ZHANG ; Liqiang ZHOU ; Sheng YANG ; Lin GUI
Chinese Journal of Oncology 2014;36(12):928-932
OBJECTIVETo analyze the clinical features, therapeutic outcome and prognostic factors of mantle cell lymphoma (MCL).
METHODSClinical data of a total of 68 patients with MCL admitted from August 2003 to June 2013 in our department were retrospectively analyzed.
RESULTSOf all the patients, the median age was 58.5 years, with marked male predominance (2.8:1), 59 patients (86.8%) were in Ann Arbor stage III/IV. 56 cases (82.4%) primarily showed lymph node involvement, 49 cased showed extranodal involvement and 19 cases (38.8%) had bone marrow involvement. Patients were followed up for 4 to 122 months with a median follow up time of 35 months. The 3- and 5-year overall survival (OS) rates were 78.5% and 64.1%, respectively. The 2- and 3-year progression-free survival (PFS) rates were 41.3% and 23.7%, respectively, and the median time to progression was 20.0 months. The overall response rate (ORR) of CHOP regimen was superior to that of intense regimens (P = 0.036). Univariate analysis showed that stage III/IV,IPI score of 3-5, expression of Ki-67 higher than 30%, elevated LDH, elevated β2-MG, blastic variant, more than 5 lymph nodes involved, and failure to chemotherapy were the negative factors. Multivariate analysis showed that Ki-67 index, LDH and the response to chemotherapy were independent factors affecting survival.
CONCLUSIONSMost patients with MCL were elderly males with advanced stage and usually had bone marrow involvement. Although ORR of CHOP regimen is superior to intense regimens, the prognosis of MCL remains poor.
Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; Cyclophosphamide ; Disease Progression ; Disease-Free Survival ; Doxorubicin ; Female ; Humans ; Lymphoma, Mantle-Cell ; diagnosis ; Male ; Middle Aged ; Multivariate Analysis ; Prednisolone ; Prognosis ; Retrospective Studies ; Survival Rate ; Vincristine