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【Objective】 To observe the efficacy of blue laser side light in the vaporization and enucleation of non-muscle invasive bladder cancer (NMIBC). 【Methods】 A retrospective analysis was performed on the data of NMIBC patients who received transurethral enucleation of bladder tumor (TURBT) using a 50 W blue laser treatment device. The efficacy and safety of surgery were evaluated by observing the application of blue laser in the treatment of bladder cancer in different locations, operation time, presence of bladder perforation and postoperative urine routine indicators. 【Results】 A total of 16 patients completed the surgery, including 5 cases with multiple bladder tumors, 9 with single bladder tumor, and 2 with lichenoid lesions. The postoperative pathological results showed all of them were NMIBC. The operation time was (10.4±4.7) min, postoperative urinalysis was (689.4±316.7)/μL, and no intraoperative bladder perforation occurred. 【Conclusion】 The lateral infusion mode of 50 W blue laser therapy device can be applied to the surgical treatment of NMIBC. The basal vaporization and enucleation of bladder cancer can be performed under direct vision conditions, and the multi-angle enucleation is suitable for bladder cancer in different positions. The tissue penetration depth of blue laser is shallow, which is conducive to the diagnosis and grading of pathological tissues.
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Objective:To establish a disk (CD) microfluidic chip detection platform for the rapid detection of CALR-1 and CALR-2 mutations in patients with cerebral infarction, and summarize its clinical application value.Methods:Based on microfluidic technology and loop mediated isothermal amplification technology, a CD microfluidic chip detection platform for simultaneous detection of CALR-1 and CALR-2 gene mutations were established, and the sensitivity, specificity, repeatability and accuracy of the platform were verified. A total of 124 patients with cerebral infarction treated in Huashan Hospital, Shanghai Medical College, Fudan University from November 2019 to March 2021 were prospectively selected into the experimental group; and 80 healthy subjects were included in the control group. The CALR-1 and CALR-2 gene mutations in anticoagulant peripheral blood samples were detected by the CD microfluidic chip. Each chip could detect 4 samples at the same time and synchronously detect 3 indexes of each sample. The detection results could be obtained after isothermal amplification for 40 min. At the same time, sequencing method was used to verify the test results, and the consistency of the results of the two detection methods was compared.Results:Using this CD microfluidic chip platform, the synchronous amplification of 3 indexes in the sample could be completed within 40 min without the need of thermal circulation, and the whole detection process of the sample could be completed within 60 min. For samples with a high concentration of target nucleic acid, typical positive signals could be visualized after amplification for 10 min, and the test results would be available within 30 minutes after receiving the samples. The detection sensitivity of CD microfluidic chip method for CALR-1 and CALR-2 mutation load concentration was 1.0% and 0.5% respectively. Nonspecific amplification was not observed for the non-target nucleic acid samples, indicating the high specificity of this method. The coincidence rates of intra and inter batch repeatability were 100% (20/20) respectively. Two samples with CALR gene mutation were found in the cerebral infarction group, both of which were CALR-1 mutations (L367fs*46). There was no CALR-1 or CALR-2 mutation in the control group. The detection results of CD microfluidic chip method were completely consistent with the sequencing verification results (100% [204/204]).Conclusions:The CD microfluidic chip method could be used for the detection of CALR-1 and CALR-2 gene mutations in clinical samples of patients with cerebral infarction. This method has the advantages of high detection sensitivity, good detection specificity, fast detection speed and high detection flux, which is helpful to clarify the etiology of patients with cerebral infarction.
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Objective:To investigate the effects of seven-step exercise rehabilitation combined with elastic resistance training on functional fitness and cardiovascular adverse events in patients with coronary heart disease (CHD).Methods:116 patients with CHD in our hospital from March 2017 to March 2019 were selected for prospective study, and they were divided into observation group ( n=58) and control group ( n=58) according to the order of admission time. The control group was given routine intervention, and the observation group was given seven step exercise rehabilitation combined with elastic band resistance training on the basis of the control group. The intervention satisfaction, incidence of cardiovascular adverse events, functional physical fitness indexes, cardiac function [left ventricular ejection fraction (LVEF), cardiac stroke volume (SV), cardiac index (CI)] and activities of daily living (BI) were compared between the two groups. Results:After intervention, the aerobic endurance, limb flexibility, lower extremity muscle strength and upper limb muscle strength were better than those in the control group ( P<0.05). After intervention, the LVEF, CI, SV, daily behavior, physiology and daily activities scores in the observation group were higher were higher than those of the control group ( P<0.05) . The intervention satisfaction of the observation group was 94.83%(55/58) , higher than that of the control group 79.31%(46/58) ( P<0.05); the incidence of cardiovascular adverse events in the observation group was 5.17%(3/58) and 10.34%(6/58) of the control group, with no statistically significant difference ( P>0.05). Conclusions:Seven-step exercise rehabilitation combined with elastic band resistance training can effectively improve the functional fitness and heart function of CHD patients, improve daily living ability and intervention satisfaction, and prevent cardiovascular adverse events.
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Objective To investigate the the operative time of two stage soft ureteroscope lithotripsy for hemorrhagic embolism after percutaneous nephrolithotomy(PCNL).Methods The clinical data of 1 patient with massive hemorrhage after PCNL who treated with superselective renal artery embolization and ureteral soft lens were analyzed.Results The patient with postoperative bleeding after PCNL was treated with superselective renal artery branch embolization,after 30d embolization,the holmium laser lithotripsy under the soft ureteroscope for ureteral calculi was performed,and the renal pelvis mucosa smooth,no bleeding and scar formation were intraoperative visible.After operation,the stone was removed basically,and the double J tube was removed 2 weeks after operation.The patient had no special discomfort and the renal function was normal.Conclusion Postoperative 14-30d is a relatively safe time to perform flexible ureteroscopic lithotripsy for super selective renal artery embolization in the treatment of patient with massive hemorrhage after PCNL.
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Objective To investigate the predictive value of baseline neutrophil-to-lymphocyte ratio (NLR) in the curative effect and prognosis of breast cancer patients treated with neoadjuvant chemotherapy.Methods The clinical data of 304 breast cancer patients received neoadjuvant chemotherapy in Beijing Cancer Hospital during 2010 were retrospectively analyzed.The correlations of baseline NLR with clinicopathological parameters of breast cancer patients were analyzed by Wilcox or Kruskal-Wallis tests,and the correlation of NLR with pathologic complete response (pCR) after neoadjuvant chemotherapy was analyzed by the binary logistic regression.The effect of clinic pathological parameters on the disease-free survival(DFS) of the patients was evaluated by the univariable and muhivariable Cox regression models.Results The medians of NLR in pCR + patients (n =62) and pCR-patients (n =242) were 1.76 and 1.72,respectively,and there was no significant difference between themn (P ＞ 0.05).Multivariate analysis indicated that the TNM stage (P ＜ 0.05) and pCR (P ＜ 0.05) were the independent risk factors influencing the prognosis of breast cancer patients.There was no significant correlation between NLR and 5-year DFS (HR=0.842,95％CI=0.566-1.255,P＞0.05).Condusion NLR lacks the predictive value in the curative effect and prognosis of breast cancer patients treated with neoadjuvant chemotherapy,while TNM stage and pCR are the independent risk factors influencing the prognosis of breast cancer patients.
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Objective To validate and compare the pyramidal tracts traced by functional magnetic resonance mo?tion activated area as the region of interest method (fMRI guided DTI-FT) and by the anatomy of primary motor cortex as region of interest method(traditional DTI-FT)using subcortical electrical stimulation (DsCS). Methods A prospective study was conducted in 12 cases of patients with central lesions involving the motor area. The pyramidal tracts were traced by fMRI guided DTI-FT method and traditional DTI-FT method. The lesions were resected with the assistance of neuronavigation. The distances between same stimulation positive point and pyramidal tracts traced by the fMRI DTI-FT or traditional DTI-FT were recorded. The coincidence rates between pyramidal tract imaging and DsCS were analyzed in order to verify the accuracy and reliability of these two methods. Results Two cases were excluded:one due to the failure of the fMRI activation caused by movement dysfunction and one case due to negative electrical stimulation.,The pyrami?dal tracts were successfully reconstructed in the rest 10 patients using these two methods which were further applied to assist surgery. The coincidence rates between DsCS and pyramidal tracts were 77%in fMRI DTI-FT and 70%in tradition?al DTI-FT. The shortest distances were 4.3mm±2.8mm and 5.5mm±3.4mm in fMRI DTI-FT and in traditional DTI-FT in 16 DsCS positive sites and the difference was statistically significant (P<0.05). Five cases had temporary postoperative pa?ralysis. Among them, four cases had upper limb paralysis and one case had hemiplegia. The motor function was improved in four cases and remained unchanged in two cases two weeks after the operation. The motor function in the rest six cases did not have any change before and after operation. Conclusion The fMRI guided DTI-FT can be helpful to deal with le?sions and effectively protect the brain function area in patients with the central area lesions involved motor area.
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<p><b>OBJECTIVE</b>To investigate the associations between physical exercise and quality of life in breast cancer patients.</p><p><b>METHODS</b>A cross-sectional study was conducted among 3 344 community breast cancer patients between April and July 2013 in Shanghai, China. Data were collected using a questionnaire, including socio-demographic situation, cancer survival and health behaviors, and scores of EORTC QLQ-C30 Simplified Chinese version and FACT-G Simplified Chinese version.</p><p><b>RESULTS</b>Among a total of 3 344 breast cancer patients, the patients doing exercise reported significantly higher EORTC physical functioning scores, role functioning scores, emotional functioning scores, global health scores, and FACT-G physical well-being scores, social well-being scores, emotional well-being scores, functional well-being scores, and FACT-G total scores than the patients who didn't take exercise (P < 0.05, P(Adjusted)<0.05) . Breast cancer patients who did exercise more than or equal to 5 times/week reported significantly higher EORTC role functioning scores, cognitive functioning scores, emotional functioning scores, global health scores and FACT-G physical well-being scores, functional well-being scores, and FACT-G total scores than patients who did exercise less than 5 times/week (P < 0.05, P(Adjusted)<0.05 ).</p><p><b>CONCLUSIONS</b>There are active associations between physical exercise and quality of life in breast cancer patients. Engagement in physical exercise is beneficial to breast cancer patients with long-term survival.</p>
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Humans , Asian People , Breast Neoplasms , Epidemiology , China , Epidemiology , Cross-Sectional Studies , Exercise , Quality of Life , Surveys and QuestionnairesABSTRACT
<p><b>OBJECTIVE</b>To investigate the associations between vegetables and fruit intake and quality of life in breast cancer patients.</p><p><b>METHODS</b>A total of 3 344 community breast cancer patients were selected through cluster sampling method between April and July 2013, in Shanghai, China. Data were collected using a questionnaire, which included socio-demographic situation, cancer survival and health behaviors(i.e. vegetables or fruit intake, exercise), European Organization for Research and Treatment (EORTC) QLQ-C30 Simplified Chinese version(3rd edition) and Functional Assessment of Cancer Therapy scale (FACT-G) Simplified Chinese version(4th edition) were used to evaluate the quality of life. Crude quality of life scores were compared between groups. Multiple linear models were used to calculate and compare adjusted means of quality of life between groups, controlling relevant factors.</p><p><b>RESULTS</b>After adjusting relevant factors, breast cancer patients who ate more than 250 g vegetables reported higher EORTC physical functioning scores, cognitive functioning scores, emotional functioning scores, global health scores than patients who ate equal or less than 250 g vegetables(respectively (80.79 ± 0.85) vs (79.34 ± 0.82), (80.07 ± 1.03) vs (77.84 ± 0.99), (84.17 ± 0.95) vs (82.76 ± 0.92), (65.75 ± 1.50) vs (62.92 ± 1.45)), t values respectively were 2.76, 3.54, 2.40, 3.17, all P values were <0.05; and breast cancer patients who ate more than 250 g vegetables reported higher FACT-G social well-being scores, function well-being scores, FACT-G total scores than patients who ate equal or less than 250 g vegetables (respectively (17.92 ± 0.40) vs (17.31 ± 0.39), (14.86 ± 0.42) vs (14.34 ± 0.40), (74.78 ± 1.01) vs (73.05 ± 0.97)), t values respectively were 2.49, 2.05, 2.90, all P values were <0.05. After adjusting relevant factors, breast cancer patients who ate fruit everyday reported higher EORTC physical functioning scores, role functioning scores, cognitive functioning scores, emotional functioning scores, social functioning scores, global health scores than patients who didn't eat fruit everyday (respectively (80.40 ± 0.82) vs (79.22 ± 0.87), (89.81 ± 1.00) vs (88.06 ± 1.05), (79.78 ± 0.99) vs (77.11 ± 1.04), (84.43 ± 0.92) vs (81.56 ± 0.97), (77.95 ± 1.25) vs (75.56 ± 1.31), (65.48 ± 1.44) vs (61.74 ± 1.51)), t values respectively were 2.15, 2.64, 4.07, 4.71, 2.89, 4.02, all P values were <0.05; and breast cancer patients who ate fruit everyday reported higher FACT-G physical well-being scores, social well-being scores, emotional well-being scores, functional well-being scores, FACT-G total scores than patients who didn't eat fruit everyday(respectively (23.35 ± 0.26) vs (22.85 ± 0.28), (17.91 ± 0.39) vs (16.98 ± 0.41), (18.59 ± 0.22) vs (18.18 ± 0.23), (14.79 ± 0.40) vs (14.17 ± 0.42), (74.71 ± 0.97) vs (72.17 ± 1.02)), t values respectively were 2.92, 3.65, 2.91, 2.35, 4.05 , all P values were <0.05.</p><p><b>CONCLUSION</b>There are active associations between vegetables / fruit intake and quality of life in breast cancer patients. Proper diet may help improve quality of life in breast cancer patients.</p>
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Female , Humans , Breast Neoplasms , China , Diet , Feeding Behavior , Fruit , Quality of Life , Surveys and Questionnaires , VegetablesABSTRACT
BACKGROUND:Superparamagnetic iron oxide nanoparticles (Fe3O4 NPs) have been widely used in MRI. It is vital to prepare the superparamagnetic MRI contrast agent with high stability, biocompatibility and tumor targeting in order to prevent the aggregation of Fe 3 O 4 NPs and realize the high-precision diagnose of tumor. OBJECTIVE:To prepare the amphiphilic superparamagnetic composite particles with tumor targeting mediated by folate receptor. METHODS:The stable amphiphilic superparamagnetic composite particles with tumor targeting function were prepared by coating the Fe3O4 NPs with a Pluronic F127-folic acid conjugate, which was synthesized via an esterification reaction between the carboxyl group of the tumor targeting molecule, folic acid and the hydroxyl group of an amphiphilic triblock copolymer, Pluronic F127. The resultant Pluronic F127-folic acid-Fe3O4 composite particles were characterized by transmission electron microscopy, Fourier transform infrared-spectra, UV-vis absorption spectra, thermal gravimetric analysis, vibrating sample magnetometer and T2-weighted imaging. WST assay was used to characterize their cytotoxicity preliminarily. RESULTS AND CONCLUSION:The Pluronic F127-folic acid conjugates were prepared via esterification reaction. Then Fe 3 O 4 NPs were wrapped with Pluronic F127-folic acid to result in the superparamagnetic composite particles with wel dispersion and biocompatibility. The size of most superparamagnetic composite particles was less than 200 nm and the size of Fe 3 O 4 core was 10-20 nm from the observation of transmission electron microscopy. The results from the Fourier transform infrared-spectra and UV-vis absorption spectroscop confirmed that folic acid molecules were modified on the surface of the superparamagnetic composite particles successful y. The mass ratio of Pluronic F127-folic acid conjugate was determined by thermal gravimetric analysis as 27.2 wt%in the resultant Pluronic F127-folic acid-Fe 3 O 4 composite particles. The saturated magnetic intensity of the superparamagnetic composite particles was 47.35 emu/g by vibrating sample magnetometer and the relaxation rate was 0.025×106 mol/s from MRI. The WST assay showed the negligible cellcytotoxicity of Pluronic F127-folic acid-Fe3O4. The superparamagnetic composite particles have potential application as the MRI contrast agents with tumor targeting, and the Pluronic F127-folic acid-Fe 3 O 4 composite particles is expected to be used as a MRI contrast agent for tumor targeting.
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BACKGROUND:It is vital to choose the appropriate carrier with low toxicity and high gene transfection efficiency in gene therapy, which is harmless to human body and environment. OBJECTIVE: To prepare superparamagnetic Fe3O4/SiO2-polyethyleneimine (PEI) composite particles. METHODS: Fe3O4 nanoparticles were prepared via an emulsion solvent evaporation method and superparamagnetic Fe3O4/SiO2 core shel microspheres were prepared successfuly subsequently via a modified stober method. The microspheres were further modified with PEI to obtain superparamagnetic Fe3O4/SiO2-PEI composite particles. The structures and properties of resultant composite particles microspheres were characterized by transmission electron microscopy, zeta potential and vibrating sample magnetometer. Superparamagnetic Fe3O4/SiO2-PEI composite particles were mixed with plasmid DNA at different mass ratios (29∶1, 39∶1, 49∶1, 59∶1, 68∶1, 78∶1, 88∶1). Thein vitro gene transfection ability was evaluated by Hela cels with the transfection of plasmid DNA encoded with green fluorescent protein and the transfection efficiency was determined by confocal fluorescence microscopy. RESULTS AND CONCLUSION: We successfuly synthesized the Fe3O4/SiO2-PEI composite particles with good dispersibility and even size distribution (about 100 nm). The surface charge was 21.07 mV, and the saturation magnetization was 28.05 emu/g that meant superparamagnetism. When the mass ratio was 59∶1, al the plasmid DNA was adherent to the Fe3O4/SiO2-PEI composite particles; when the mass ratio was > 59∶1, there were excessive Fe3O4/SiO2-PEI composite particles. Therefore, the mass ratio of 59:1 could lead to a better outcome for HeLa celltransfection. These results indicate that the Fe3O4/SiO2-PEI composite particles can dramaticaly improve the transfection efficiency of plasmid DNA compared with PEI.
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Objective To explore the preliminary mechanism of mesenchymal stem cells (MSCs) in promoting prostate cancer proliferation in tumor inflammatory microenvironment.Methods From April 2013 to October 2013,MSCs pretreated with inflammatory cytokine IL-1α (MSCs (IL-1α)) and its culture supernatants mixed with RM-1 cells,which origined from C57BL/6 mice,were subcutaneously administered in the armpit area of C57BL/6 or BALB/c mice to establish homologous or heterologous transplant animal mode and to detect the tumor growth.Meanwhile the influence of MSCs on the proliferation of spleen cells was detected in vitro.Results In homologous transplant model,the relative tumor weight of prostate cancer cells prtreated with MSCs and MSCs (IL-1α) and their culture supernatant were (3.4 ± 0.2),(3.3 ±0.2),(4.9±0.5),and (5.2±0.6) g.The results were statistically significant (P＜0.05) compared with the control group (2.4±0.2) g.In heterologous model,the ratio of tumor formation of the pretreated groups were 50％,50％,80％ and 80％,respectively,compared with the control group of 0％.The results were statistically significant (P＜0.05).In proliferation experiments of spleen cells,the number of spleen cell pretreated with IL-1α were significantly lower than that in control group and unpretreated group (P ＜ 0.05).Conclusions MSCs pretreated with IL-1α could effectively promote the growth of prostate cancer cell in vivo.The reason may be due to inflammatory cytokines induce immune suppression of MSCs and then lead to immune escape of cancer cells.
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Objective To establish a SYBR green Ⅰ real-time PCR method for detecting serum miR-21 and preliminarily explore its value in diagnosing breast cancer.Methods Total RNA was extracted from serum by Trizol reagent.Then miR-16 ( internal reference gene for miR-21 ) and miR-21 were reverse transcribed into corresponding cDNA by stem-loop RT primers.Their cDNA were amplified and detected by using SYBR green Ⅰ real-time PCR.The accuracy of assay was analyzed by signal to noise ratio (SNR) ; the specificity of assay was evaluated by melting curve; the precision of assay was assessed by R2 of standard curve and the stability of assay was calculated by intra-assay and inter-assay variation.Furthermore,the level of miR-21 and miR-16 were detected by this method among the serum samples of 33 breast cancer patients,18 benign breast disease patients and 49 healthy individuals.And the sensitivity and specificity in breast cancer diagnosis were evaluated according to cut-off value which was defined by relative expressions of miR-21 between breast cancer patients and healthy population.Results Through optimization of temperature and time in the annealing and extension stage during PCR,SNR was ≥99.36％ ; peak of melting curve was single; R2 of standard curve was 0.994 8 and Coefficient of Variance (CV) of intra-assay ＜ 1.5％,CV of inter-assay ＜4％.They indicated that this method was accurate,specific,precise and stable.When miR-16 was taken as internal reference,the relative expressions of serum miR-21 detected by SYBR green Ⅰ realtime PCR among the serum samples of breast cancer patients,benign breast disease patients and healthy population were 20.83 ± 18.18,20.86 ± 10.11 and 9.33 ±4.44,which had statistical significance among of them (x2 =16.92,P ＜ 0.001 ).There was statistical significance in healthy people vs breast cancer patients ( Z =- 2.58,P ≤ 0.01 ) and healthy people vs benign breast disease patients ( Z =- 4.42,P ≤0.01 ),but was not between breast cancer patients and benign breast cancer patients (Z =-0.51,P =0.608).When the value of 18.32 for the relative expressions of miR-21 was defined as cut-off value,the sensitivity and specificity of this method to diagnose breast cancer were 51.5％ (17/33)and 93.9％ (46/49),respectively.Conclusions A sensitive,specific and stable SYBR green Ⅰ real-time PCR for detecting serum miR-21 has been established.This method may have some diagnostic value for breast cancer.
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ial of cells. It may serve as an index for monitoring and prognostic diagnosis of breast cancer.
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Objective To investigate the feasibility of screening for esophageal cancer with survivin colloidal gold immunochromatographic test strip. Methods The serum samples from 158 esophageal cancer patients and 146 healthy individuals were tested with survivin colloidal gold immunoohromatographic test strips. Results 20 nm-diameter colloidal gold solution was prepared and survivin polyclonal antibodies with different concentrations were labeled to the gold particles. It showed that the optimal concentration for labeling was 12 μg/ml. The prepared survivin colloidal gold immunochromatographic strip teat showed positive rates of survivin in esophageal cancer group and control group were 51.9 % (82/158) and 15.1% (22/146) respectively (χ~2 = 45.7, P < 0.05). No statistical differences were observed in age groups (between 42-54 years: 47.1%; between 56-68 years: 40.9%; between 69-81years: 63.9%, χ~2= 1.11, P > 0.05), gender groups (male: 50.4%, female: 58.1%, χ~2= 0.59, P > 0.05) , primary site groups (upper segment: 56.3%, middle segment: 46. 1%, lower segment: 32. 1%, χ~2 = 2.64, P > 0.05), differentiation groups (well-differentiation: 56.3%, moderate differentiation: 43.2%, poor-differentiation: 32.7%,χ2= 1.63, P >0.05)and lymph node metastasis groups (with metastasis:43.3%, without metastasis:43.4%, χ~2 = 0.00, P > 0.05). The sensitivity, specificity and accuracy of survivin colloidal gold immunochromatographic test strip was 51.9% (82/158), 84.9% (124/146) and 67.8% (206/304) respectively. Conclusions Survivin colloidal gold immunochromatographic test strip is fast, simple, easy-to-read. It could be used as a valuable tool for screening of high-risk patients with esophageal cancer.
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Objective: To investigate the possible association between the allelic variation of LAPTM4B and the genetic susceptibility of lung cancer. Methods: The genotype of LAPTM4B was analyzed in 134 unrelated healthy adult individuals and 166 patients with lung cancer by utilizing polymerase chain reaction based on special primers. The genotypical distribution of LAPTM4B was analyzed by ? 2 test. Results: The allelic frequencies of the *2 were 40.1% and 28.0% in the lung cancer group and the healthy control group respectively, which was significantly different between the two groups (P 0.002). There was a significant difference in the overall genotypical distribution between the patients and the controls (P0.005). The risk of suffering from lung cancer was increased 1.91 times in the individuals of the *1/2 genotype (95%CI: 1.178-3.110) and 3.26 times in the individuals of the *2/2 genotype of LAPTM4B (95%CI:1.338-7.929) compared with the *1/2 genotype. No association was observed between the genotypical distribution of LAPTM4B and the clinical information on patients of lung cancer such as gender, age, pathological type, differentiation classification of TNM and infection of HBV. Conclusion: This study suggests that the allele *2 of LAPTM4B might be the risk factor of lung cancer, which could be associated with genetic susceptibility of lung cancer.
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Tumor was one of the most threatening disease of the human beings. Tumor marker plays a very important role in tumor diagnosis, estimating curative effects and prognosis. Reasonable usage of detection technology for tumor marker has direct effect on clinical tumor diagnosis and treatment. We were make a brief summary to actuality of the present applying tumor detection technologies in clinic and their developmental trends.
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<p><b>OBJECTIVE</b>To develop a tumor imaging agent for vasoactive intestinal peptide (VPAC) receptor and evaluate its biological activity and pharmacokinetics of radiolabeled peptide.</p><p><b>METHODS</b>VIP(28) was modified at the carboxyl terminal by the addition of His-tag which was the chelating site of (99m)Tc(I) and the general purification tag for immobilized metal ion affinity chromatography. Biological activity of the modified VIP(28) analogue MY34 was examined in vitro by radiological cell-binding assay, rabbit internal anal sphincter (IAS) smooth muscle relaxing assay and immunocytochemical stain. The pharmacokinetics of this labeled peptide was examined in C57 mice.</p><p><b>RESULTS</b>MY34 could relax the IAS smooth muscle and bind VPAC receptors on tumor cell membranes. (99m)Tc- MY34, with a yield of about 90%, was stable enough for practical use. Both MY34 and VIP(28) could inhibit the binding between the labeled peptide and VPAC receptor. The pharmacokinetics of [(99m)Tc(H(2)O)(3)(CO)(3)]-MY34 was studied in mice conformed well with the two-compartment model (Wi = 1/C(2)), with a t(1)/(2alpha) of 16.35 min and a t(1)/(2beta) of 1013.56 min.</p><p><b>CONCLUSION</b>MY34 possesses physiological activities and specific receptor binding characteristics similar to those of natural VIP(28).</p>
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Animals , Mice , Rabbits , 3T3 Cells , Binding, Competitive , Isotope Labeling , Mice, Inbred C57BL , Muscle, Smooth , Physiology , Organotechnetium Compounds , Peptides , Metabolism , Pharmacology , Radionuclide Imaging , Receptors, Vasoactive Intestinal Peptide , Stomach Neoplasms , Tumor Cells, Cultured , Vasoactive Intestinal Peptide , Metabolism , PharmacologyABSTRACT
Objective To clone human neuron-specific enolase (NSE)gene and prepare the monoclonal antibodies against human neuron-specific enolase and to test the expression of NSE in tumor cell lines by immunocytochemistry (ICC).Methods The gene fragment of human NSE was amplified by RT-PCR and ligated to the pGEM vector. After the sequencing of recombinant NSE, it was ligated to the expression vector pMS-31b. The MS2-NSE fusion protein was expressed after higher temperature induction. The purified target protein was used for immunizing BALB/C mice to prepare McAbs against NSE.Results Full length of NSE gene with 1 305 bp was cloned. Molecular weight of MS2-NSE was 57 000. 1.42 mg/L of MS2-NSE fusion protein could be expressed. Two strains of hybridoma secreting NSE McAbs were obtained by ELISA screening. The subtypes of the NSE McAbs were IgG1and IgG2a. The two McAbs could react with A549 cell lines in ICC. NSE positive staining in ICC was mainly located in cytoplasm.Conclusions We clone human neuron-specific enolase gene, obtain the anti-NSE monoclonal antibodies and examine the expression of NSE in lung cancer tumor cell line.
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Objective To clone survivin gene, prepare its monoclonal antibody(McAb) and check its expression in liver carcinoma cells.Methods Survivin gene cDNA was amplified by RT-PCR from human breast carcinoma cell line MCF-7 and constructed into prokaryocytic expression vector.Fusion-protein of human Survivin was expressed and used for immunizing BALB/C mice.The spleen cells from immunized mice were fused with SP2/0 cells and selectively cultured with HAT medium.ELISA and Western blot were used to screen and identify the McAbs.Immunocytochemical staining was applied for survivin expression in liver carcinoma cells.Results The full survivin gene was cloned. The hybridoma cell that secret specific monoclonal antibody against human surviving was identified.The immunoglobulin subclasses of the McAbs were IgG1.Western blot showed that the McAbs against survivin can specifically react with MS-Survivin fusion protein. The positive reaction was found in hepatocarcinoma cell line HepG2 and hepatocarcinoma tissue by immunocytochemical staining.Conclusion The McAbs against the human Survivin were successfully prepared by a MS2-Survivin fusion protein expressed by E.coli. and the McAb had positive reaction with HepG2 and hepatocarcinoma tissue. It may be a useful tool to study the functions of survivin and check clinical cancer samples.
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Objective To clone human survivin gene and express survivin-an inhibitor of apoptosis proteins in Pichia pastoris eukaryotic expression system. Methods Full length survivin gene was amplified by PCR using survivin specific primers. The verified survivin gene by sequencing was subcloned into pPic9k. The recombinant plasmid was linearized by restriction enzyme cutting. The linear survivin gene was introduced into GS115/ His-cells by electroporation. The transformants were transferred onto YPD plates that contained different concentrations of G418 for screening the positive clones. The integrated survivin gene in positive clones was confirmed by PCR. Selected transformants were cultured in BMMY medium with 1 % methanol for inducing the expression of survivin protein. The expressed survivin protein was analyzed by ELISA. Results The cloned human survivin sequence was the same as that in the GeneBank. The recombinant pPic9k-survivin was constructed in Picha pastoris eukaryotic expression vector. After the linear digestion, the recombinant vector was introduced into GS115/ His-cells, the 6 positive clones against G418(4 mg/mL) were obtained and confirmed by PCR; the highest survivin protein was expressed when expressed for two days in 1 % methanol BMMY medium. Conclusions Improved survivin protein yield may be reached by modifying the experimental conditions. This will help to further study the biological functions of survivin and its roles in tumor developments.