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Objective:To observe the effects of Yikunjing capsules on the bone histomorphometry indexes and the serum Interleukin-6 (IL-6) level in mice with osteoporosis.Methods:60 cases ofC57 female mice were randomly divided into 5 groups,such as model group (equal volume of normal saline),estrogen group (nilestriol,0.25 mg/kg),Yikunjing capsules high dose group (1.44 g/kg),Yikunjing capsules medium dose group (0.72 g/kg) and Yikunjing capsules low dose group (0.36 g/kg),sham group (equal volume of normal saline) were treated with sham operation.The mice in each group was intragastrically administrated for 70 days.Enzyme linked immunosorbent assay (ELISA) was used to detect the serum IL-6 level.The bone histomorphometry index were detected with BI-2000 Medical Image Analysis System.And the Number of blood vessels in distal femoral metaphysis of each group was measured by CT.Results:Compared with the sham group,the width and area of trabecular bone,the thickness of cortical bone,area of trabecular bone,the number of osteoblasts,the bone mineral density and the number of blood vessels in model group were significantly reduced(P＜0.05),while the number of osteoclasts and serum IL-6 level were significantly increased (P＜0.05).Compared with the model group,above indexes in estrogen group,Yikunjing capsules high,medium,low dose group were improved (P＜0.05),and the effect of Yikunjing capsules high dose group and estrogen group was almost the same (P＞0.05).Conclusion:Yikunjing capsules can rectify the bone histomorphometry indexes reduce the serum IL-6 level and increase the number of blood vessels in mice with osteoporosis,and intragastrically administrating 1.44 g/kg Yikunjing capsules could get the better effect.
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Objective To observe the characteristics of different blood perfusion in renal cell carcinoma by contrast-enhanced ultrasound and prove its diagnostic value.Methods One hundred and six cases of renal cell carcinoma confirmed by patholog were involved,and were divided into three groups:clear cell carcinoma group,papillary cell carcinoma group,and chromophobe cell carcinoma group.Their imaging characteristics were observed.Time-intensity curve was analyzed by using QLab software.The difference of time of beginning to increase (△AT =ATrenal cell carcinoma-AT surrounding normal renal parenchyma) and the difference of peak intensity(△PI =PIrenal cell carcinoma-PIsurrounding normal renal parenhyma) in three groups were compared.Results △AT and △PI in clear cell carcinoma group were different from those in the papillary cell carcinoma group and chromophobe cell carcinoma group (P ＜0.05).The difference between papillary carcinoma group and chromophobe carcinoma group was not significant (P ＞0.05).Contrast agent filled very fast in most clear cell carcinomas,filled speed and time to peak were early or synchronously with the surrounding renal tissue.Compared with renal papillary carcinoma and chromophobe carcinoma group,clear cell carcinoma group showed a high peak intensity and part of the surroundings appeared high increasing signs,like of a ring.Contrast agents filled slow in papillary carcinomas group and chromophobe carcinomas group,and the peak intensities were low.Conclasions Contrast enhanced ultrasound can provide important clinical value for the diagnosis of renal cell carcinoma and subtype diagnosis.
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Objective To summarize the relationship between metabolic syndrome (MS),its components and T1 stage with high grade urothelial carcinoma (HGUC) of the Bladder.Methods The clinical data of 200 patients with T1 high grade bladder cancer who were admitted to our hospital from January 2010 to June 2014 were retrospectively analyzed,including 155 males and 45 females.Ages were 24 to 86 years old,average 66 years old.Based on the history or blood glucose levels,patients were divided into diabetic group (n =41) (20.5％) and non diabetes group 159 cases (79.5％);According to the body mass index (BMI) were divided into obese group (≥25 kg / m2) of 98 cases (49.0％) and non obese group (＜ 25 kg / m2) of 102 cases (51.0％).According to the blood pressure level,71 cases (35.5％) were divided into hypertension group and 129 cases of non hypertension group (64.5％).MS and its components and the relationship between the recurrence and progress of bladder cancer were analyzed.The Kaplan Meier method was used to assess MS and its components division of tumor progression free survival (progress-free survival,PFS) and recurrence free survival (recurrence-free survival,RFS) influence.Cox regression model of multi factor analysis were used to evaluate the PFS and RFs of MS and its components with bladder cancer.Results Of the 200 cases,16 cases (8.0％) were MS.Tumor recurrence occurred in 121 cases (60.5％),and 84 patients (42.0％) were in progress.Diabetes and non diabetes groups the average RFs were 21.7 and 29.3 months respectively,and the difference was statistically significant (x2 =10.115,P =0.001);The median PFS were 32.8 and 39.8 months respectively,the difference has statistical significance (x2 =14.760,P ＜0.001).Obese group and non obese group average RFs were 34.7 and 42.0 months respectively,and the difference were statistically significant (x2 =16.077,P ＜ 0.001);The median PFS were 22.8 and 32.6 months respectively,the difference was statistically significant (x2 =16.174,P＜0.001).The average RFS of MS group and non MS group were 21.5 and 28.4 months respectively,the difference was statistically significant (x2 =5.429,P =0.02);the average PFS was 35.1 and 38.7 months respectively,and the difference was statistically significant (x2 =3.854,P ＜ 0.05).Cox multivariate survival analysis showed that diabetes and obesity can increase the risk of recurrence and progression of T1 advanced stage bladder cancer (HR =1.792,P =0.013,HR =2.498,P ＜ 0.001;HR =0.559,P ＜ 0.001;HR =0.492,P ＜ 0.001).Conclusions Diabetes mellitus and obesity are high risk factors for the recurrence and progression of T1 advanced stage bladder cancer,but MS is not related to the prognosis of T1 patients with advanced bladder cancer.
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Objective To evaluate the value of digital subtraction computed tomography angiography(DSCTA)in the diagnosis of intracranial small aneurysms.Methods 92 patients with intracranial aneurysms confirmed by digital subtraction angiography(DSA) or surgery were selected as the subjects of this study.Their clinical and CT imaging data were analyzed retrospectively.The diagnos-tic accuracy of DSCTA and conventional CTA for intracranial aneurysms was compared.Results A total of 102 aneurysms were con-firmed in the 92 patients,100 of them were detected by DSCTA and 89 by conventional CTA (χ2 =8.707,P =0.003).Two small aneurysms of less than 5.0 mm located respectively at the cavernous sinus and beneath the clinoid segments were missed by DSCTA, while 13 by conventional CTA.When aneurysms of less than 3.0 mm and 3.0-5.0 mm were combined with calculation,the sensi-tivity of DSCTA in detecting aneurysms of less than 5.0 mm was significantly higher than that of conventional CTA (χ2 =8.393,P =0.004).Conclusion DSCTA has more advantages than conventional CTA in the diagnosis of intracranial aneurysms,especially for the small aneurysms adjacent to the skull base.DSCTA can be used as a preferred technique in the screening and diagnosis of in-tracranial aneurysms.
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Objective To observe the effect of combination of dexmedetomidine and sufentanil on postoperative analgesia for the highly nicotine dependent patients undergoing esophagectomy. Methods Ninety highly nicotine dependent patients undergoing esophagectomy were randomly allocated into three groups (n = 30 each): low-dose sufentanil group (Group S1), high-dose sufentanil (Group S2), dexmedetomidine and sufentanil group (Group DS). All patients of three groups received postoperative PCIA with following setting: 2 mL/h;bolus: 2 mL; lock time: 5 min; 4 hours limited: 40 mL. The scores of VAS and Ramsay, effective presses/total presses of PCIA, the consumption of sufentanil during 72 h after operation, side effects and the satisfaction degree of patients were recorded. Results Compared with those of group S2, the scores of VAS (both at rest and movement) decreased significantly in group DS from 1 h to 8 h (P<0.05). Compared with those of group S1, the scores of VAS (both at rest and movement) decreased significantly in group S2 and DS from 1 h to 72 h (P<0.05). Compared with that in group S1 and S2, the consumption of sufentanil during 72 h after operation decreased significantly in group DS (P < 0.05). Compared with those in group DS, The numbers of nausea and vomiting were significant decreased in group S1 and S2(P < 0.05). Compared with those in group S1, the remedial cases were significantly decreased in group S2 and group DS (P < 0.05). Conclusion The effect of combination of dexmedetomidine and sufentanil is superior to that of sufentanil in terms of postoperative analgesia in highly nicotine dependent male patients undergoing esophagectomy.
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Objective To evaluate the relationship between metabolic syndrome , its components and the histopathological findings in bladder cancer patients .Methods The data of 326 patients in our department between October 2010 and October 2013 were retrospectively analyzed.Age, gender, stature, weight, histologic stage, grade, and the presence of hypertension , diabetes mellitus, body mass index ( BMI) were evaluated.There were 64 females, 262 males, aged 23-89 years, including 241 low stage, 85 high stage, 155 low grade, and 171 high grade, respectively.There were 117 cases with hypertension, 95 cases with diabetes mellitus , 139 cases with BMI ≥25 kg/m2 and 49 cases with metabolic syndrome.The TNM classification was used , with Ta and T1 tumor accepted as low stage , T2 , T3 and T4 tumor as high stage bladder cancer.In addition, the pathological grading system adopted by the 2004 World Health Organization was applied.Non-invasive papillary urothelial neoplasms of low malignant potential were regarded as low grade.Analyses were completed using Chi-square tests to evaluate the correlation of diabetes mellitus , hypertension and obesity with the pathologic stage and grade .Moreover , the pathologic stage , grade and recurrence were compared between metabolic syndrome and non-metabolic syndrome groups . Results Metabolic syndrome was significantly associated with histological grade and stage (P=0.001, P=0.011). Diabetes mellitus and obesity were also associated with histological grade and stage (P=0.006, P<0.01). Conclusions Patients with metabolic syndrome were found to have significant higher T stage and grade of bladder cancer .Diabetes mellitus and obesity may promote the grading and staging of bladder cancer .
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Objective To investigate the clinicopathological features, treatment modalities, and prognostic factors for survival in patients with urinary tract small cell carcinoma (UT-SCC).Methods A total of 25 patients treated from June 2000 to December 2014 were included in the retrospective study.The data included age, gender, primary tumors origins, stage, treatment modalities, progression-free survival (PFS), overall survival (OS), pathology and immunohistochemistry.Of these cases, 22 were male, and the other was female, whose age was 45-79 years (mean age 67).20 cases small cell carcinoma of bladder patients and 2 small cell carcinoma of prostate cancer patients were included.The number of small cell carcinoma in pelvis,ureter and retroperitoneal was 1 respectively.The patients with small cell carcinoma of the urinary tract were classified as disease and extensive disease.17 bladder small cell carcinomas were limited disease and 3 cases were extensive disease;Prostate small cell carcinomas were both extensive disease;The small cell carcinomas in pelvis, ureter were limited disease;The small cell carcinoma in retroperitoneal was extensive disease.10 bladder small cell carcinomas which were limited disease received radical cystectomy.6 of 10 patients received etoposide and cisplatnum (EC).4 of 10 patients received gemcitabine and cisplatnum (GC).7 bladder small cell carcinomas patients who with limited disease refused to receive radical cystectomy in which 2 patients received TURBT and 5 patients received TURBT followed chemotherapy.Both prostate small cell carcinomas received chemoradiotherapy.2 small cell carcinomas in upper urinary tract (pelvis and ureter) received radical nephroureterectomy with bladder cuff resection.The patient of retroperitoneal small cell carcinoma received percutaneous nephrostomy after biopsy.The progression-free survival (PFS) and overall survival (OS) of these patients are analyzed;the influence of TURBT with adjuvant chemotherapy and clinicopathologic characteristics were analyzed in median PFS and OS.PFS and OS were compared between groups as a function of time, using a Kaplan-Meier survival curve analysis and the log-rank significance test.All statistical tests were two-sided, and P values ＜ 0.05 were considered statistically significant.Results 25 patients with a pathologic confirmation of UT-SCC,either by biopsy or surgery,were finally included.These patients were classified as pure UT-SCC (14) and Mixed UT-SCC (11).Mixed UT-SCC was defined as tumors containing both SCC and non-SCC components,regardless of the proportion of the latter.13 cases were strongly positive and 3 cases were weakly positive in neuron specific enolase (NSE) level.8 cases were strongly positive and 2 cases were weakly positive in CgA level.Patients with limited disease experienced a significant longer PFS and OS compared with extensive disease subjects (PFS 13.2 vs.7.8 x2=13.53 P＜0.01;OS27.2 vs.12.7x2=19.88 P＜0.01).Patients with bladder SCC showed a significantly higher median PFS and OS compared with patients with SCC of other parts of urinary tract (PFS 12.8 vs.8.2 x2 =12.00, P =0.001;OS 26.3 vs.13.2 x2 =14.45,P ＜0.01) .The two different chemotherapy regimens (GC and EC) have no influence on survival (PFS: 16.3 vs.12.5,x2 =3.34, P =0.07;OS 29.5 vs.22.8, x2 =1.66, P =0.198).TURBT followed by adjuvant therapy have no influence on survival (PFS 14.5 vs.12.0 t =1.30 P =0.251;OS 24.5 vs.28.4 t =0.50,P =0.636).Conclusions The primary tumors origins and stage may have influence on survival in patients with UT-SCC.Patients with bladder small cell carcinoma and limited disease experienced a longer survival.
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Tumor-derived heat shock protein-peptide complex 96 (HSPPC-96) containing tumor antigenic peptides can elicit po-tent tumor-specific and protective immunity. Autologous HSPPC-96 vaccine has been shown to effectively prolong recurrence-free sur-vival and increase the overall survival of many tumors, thereby suggesting extensive future applications. However, as an autologous tu-mor-derived individual vaccine, the development of HSPPC-96 vaccine is challenged by the lack of an adequate autologous tumor, lim-ited efficacy for advanced-stage cancer, etc. This paper summarized the progress, future perspectives, and challenges in the clinical de-velopment of HSPPC-96 vaccine immunotherapy.
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Objective To evaluate the functional changes of visual cortex (V1) in patients with primary open angle glaucoma (POAG) by fMRI retinotopic mapping technology. Methods Fifteen POAG patients and 15 healthy volunteers underwent stimulations with fMRI retinotopic mapping stimulus and contrast-reversing checkerboard patterns stimulus on a Siemens Trio 3.0 T MRI whole-body scanner for functional data collection. Comparisons of V1 fMRI responses between the glaucomatous eyes and the healthy eyes of the patients were carried out using paired samples t-test, while independent samples t-test was used to compare V1 fMRI responses and activations between the healthy eyes of patients and the age-, gender- and side- matched eyes of normal people. Differences of V1 cortical functions and visual functions were analyzed by linear correlation analysis when the glaucomatous and the healthy eyes were simulated individually. Results (1) V1 fMRI responses of the individually stimulated glaucomatous eyes[(1.24±0.72)%]were weaker than those of the healthy eyes[(2.18±0.93)%](t=4.757,P<0.01). Comparisons of V1 fMRI responses between the glaucomatous eyes and matched eyes of normal people, as well as between the healthy eyes of patients and the matched eyes of normal people, were performed respectively: the responses in the glaucomatous eyes[(1.24±0.72)%]were weaker than those in the matched eyes of normal people[(2.01±0.65)%](t=-3.011,P<0.01). There was no statistical difference of the responses between the healthy eyes from patients[(2.18±0.93)%]and the matched eyes of normal people[(1.95±0.75)%](t=0.742,P＞0.05). (2) Differences of V1 cortical functions were negatively correlated with those of visual functions in the individually stimulated glaucomatous and healthy eyes (r=-0.887, P<0.01). (3) The activated area indexes of V1 cortexes in the healthy eyes from patients (0.72±0.12) were lower than those in the matched eyes of normal people (0.85±0.09) (t=-3.801, P<0.01). Conclusion Cortical function impairment was in accordance with visual function impairment in glaucoma. Located and quantified measurement with fMRI retinotopic mapping was a useful method for clinical follow-up and evaluation of functional alteration of glaucomatous visual cortex, and a potentially useful means of studying trans-synaptic degeneration of visual pathways of in vivo glaucoma.
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Objective To explore the normal range of the fourth ventricle volume of Chinese adults of the Han nationality and provide morphological data for the construction of database for Chinese Standard Brain.Methods This is a clinical multi-center study.One thousand Chinese healthy volunteers (age range= 18 to 70) recruited from 15 hospitals were divided into 5 groups, i.e., Group A (age range = 18 to 30),B (age range =31 to 40), C (age range =41 to 50), D (age range =51 to 60), and E (age range =61 to 70).Each group contained 100 males and 100 females.All of the volunteers were scanned by MR using T1 weighted three-dimensional magnetization prepared rapid acquisition gradient echo sequence.After three dimension data reconstruction, the volumes of the fourth ventricle were measured at sagittal view by automatic trace of Midobl.2 combined with manual outlining.The difference of volumes of the fourth ventricle between male and female were analyzed by independent sample t test, and among age groups by ANOVA.Pearson's correlation coeffcient was used to characterize the relationship between volumes of the fourth ventricle and age.Results The fourth ventricle volumes of Group A-E were (2.1±0.9), (2.1±0.8), (2.2±0.8), (2.1±1.0) and (2.4±0.8) ml respectively for male; those for female were(2.0±0.7), (1.9±0.6), (18±0.6), (1.9±0.7) and (2.0±0.6) ml respectively.The fourth ventricle volumes of males were significantly larger than those of females ( t = 5.573, P =0.000 ) ; there were no significant differences among the female groups ( F = 1.788, P = 0.130 ) ; there were significant differences among the male groups ( F = 2.639, P = 0.033 ) and multiple comparison found that the 60 years old was the watershed with significant difference ( P < 0.05 ).Correlation between the change of males' volumes and the ages was not strong (r = 0.119, P = 0.008 ), and the females' volumes did not correlated with their ages ( r = 0.041,P = 0.360 ).Conclusion There are gender differences in the fourth ventricle volumes of normal Chinese adults, and changes of the fourth ventricle volume with aging are different between males and females.
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Objective To explore the morphological characteristics of optic tracts in healthy Chinese Han adults on the high-resolution MRI and fill the database of Chinese standard brain with morphological data of optic tracts.Methods Cerebral MRI scans with T1 WI 3D MPRAGE sequence of 1000 healthy Chinese volunteers from 15 hospitals were divided into five stages, ranging in age from 18 to 70.With the technique of multi-baseline, structure and morphology of optic tracts were displayed optimally on the images with multiplanar reconstruction.Data were measured as following: transverse distance of the cisternal optic tract (TD1) and peri-crural optic tract (TD2), length from the cisternal optic tract to the peri-crural optic tract (L) ,angle between optic tract(AOT) and height of optic tract from its first segment to plane of anterior commissure (H) including H1, H2, H3, H4 and HS.The measurements of optic tracts between sexualities and among age groups were compared by anasis of covariance; those among five age groups were compared pairwisedly by least significant difference analysis (LSD); and the differences of measurements between left and right optic tracts were analyzed using paired t test.Results (1) Comparisons of optic tract structures between male and female: the mean optic tract length of male [(11.69±1.45),(11.56±1.44) mm] was significant longer than that of female [(10.58±1.29),(10.40±1.34) mm] (F=22.236, 29.703, P=0.000); the mean H1 of male [(2.56±0.28),(2.60±0.29) mm] and female [(2.57±0.31 ), (2.63±0.32) mm] were significantly different ( F =11.130,7.805, P = 0.000, 0.005).No significant differences of the other measurements were found between male and female ( P > 0.05 ).(2) Comparisons among age groups: among 5 age groups, TD1 of both sides [left TD1 :(4.64±0.51 ), (4.64±0.57), (4.55±0.58), (4.39±0.53), (4.36±0.58)mm;right TD1 :(4.84±0.53) ,(4.80±0.60), (4.77±0.65), (4.60±0.59), (4.57±0.59) mm] and the right TD2[(3.33±0.45),(3.34±0.41),(3.33±0.36),(3.23±0.38),(3.23±0.39) mm] had statistical differences ( F = 3.458, 2.735, 4.711, P = 0.008,0.028, 0.001 ).The LSD analysis found that the 50 years old group was the watershed with significant differences ( P < 0.05 ).The results also showed that the L of both sides had statistical differences among 5 age groups ( F = 14.510,14.532, P = 0.000).The LSD analysis found that the 60 years old group was the watershed with significant differences (P <0.05).Left TD2 and the H1-H5 of both sides had no significant differences among age groups ( P >0.05).(3) Comparisons of the measurements between left and right optic tracts: TD1 of bilateral optic tracts were (4.52±0.57)and(4.72±0.60) mm respectively; H1 of bilateral optic tracts were (2.56±0.30)and (2.61±0.30) mm respectively; H2 of bilateral optic tracts were (2.66±0.30)and (2.70±0.30) mm respectively; and L of bilateral optic tracts were ( 11.14±1.47 ) and ( 10.98±1.50 )mm respectively.There were significant differences in these measurements between left and right optic tracts( t =12.460, - 6.013,5.595,4.784, P = 0.000 ), while there were no significant differences in TD2, H3, H4 and H5( P > 0.05 ).Conclusions With high-resolution MR1 and 3D reconstruction, optic tract can be displayed clearly and measured accurately.There are definite differences in anterior segments of optic tracts between sexualities, sides and among ages in normal Chinese Han adults, while the posterior segments of optic tracts keep stable.Normal reference values of optic tracts in Chinese Han adults are provided to clinical practices and scientific researches, which are valuable for building of Chinese standard brain.
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Objective To investigate the effect of high-dose creatine phosphate (CP) on myocardial ischemia-reperfusion (I/R) injury in patients undergoing cardiac valve replacement. Methods Two hundred and forty-six ASA Ⅱ or Ⅲ patients aged 42-71 yr weighing 45-80 kg undergoing mitral-aortic valve replacement were randomly assigned into 2 groups: control group ( n = 122) and CP group ( n = 124). CP 10 g in 100 ml normal saline (NS) was infused over 60 min starting from the beginning of operation in group CP. In control group NS 100 ml was infused instead of CP. Blood samples were collected before anesthesia and on 1st and 5th postoperative day for determination of serum CK, CK-MB and LDH activity and cTnI concentration. The number of patients receiving dopamine and adrenaline at the time of return of spontaneous heart beat and 12, 24 and 48 h after operation was recorded. The incidence of arrhythmia (auricular fibrillation, ventricular arrhythmia), myocardial infarct and left ventricular ejection fraction (LVEF) were also compared between the 2 groups. Results Serum CK, CK-MB and LDH activity and cTnI concentration on 1st and 5th postoperative day, the doses of dopamine and adrenalin, the incidence of arrhythmia and myocardial infarct were significantly lower in group CP than in control group. The postoperative LVEF and the incidence of spontaneous recovery of spontaneous heart-beat were significantly higher in group CP than in control group. Conclusion Pretreatment with high dose CP can protect myocardium against I/R injury in patients undergoing cardiac valve replacement under CPB.
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Objective:To obtain antibodies against amylin from a 'naive' human Fab fragment antibody phage diasplay library and to analyze the specificity of antigen binding activity.Methods:Panning and screening Fab antibody from the antibody library,the positive clones with well reactivity to amylin were selected after five times selection of 'adsorption-elution-enrichment'.Then the plasmid DNA which was extracted from the clones,was digested with Spe Ⅰ and Nhe Ⅰ to delete gⅢ (about 660 bp).The digested 47 000 bp DNA which was purified after separation of bands from agarose gel was ligated with T4-DNA ligase.The constructed expressing phagemids were transformed to the BL21(DE3)pLysS,soluble Fab was expressed in it by the induction of IPTG and its characteristics and specificity were determined by ELISA and Western blot.Results:Soluble Fab antibodies were expressed in E.coli.According with molecular weight of IgG Fab,protein band of about 47 kD was shown by SDS-PAGE.Western blot using the goat anti human IgG-HRP showed their binding activities.ELISA showed their specificity with amylin antigens and they did not react with bovine serum albumin.Conclusion:The high level expression and identification of the soluble human anti- amylin Fab fragment antibodies has been obtained successfully,which lays a solid foundation for further researching about the biological and pathological activities of amylin.
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Objective To investigate the distribution of killer immunoglobulin-like receptor(KIR) genes in Qingdao Han population and its difference from other populations.Methods The KIR genotypes of 81 cascs were examined by PCR-SSP,and the gene frequencies were calculated and compared to those of other populations.Result Fourteen KIR genes and 2 pseudo genes were found and all had different gene frequencies.Conclusion All inhibit-gene frequencies are higher than their active-gene frequencies.KIR gene frequencies are different between the southern and northern Chinese Han people.Many of them increase gradually from the south to the north.The KIR gene distributions of Korean and Japanese populations are close to the Northern Chinese Han population.
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Background and purpose:To investigate the induction of apoptosis by epigallocatechin-3-gallate(EGCG) in xenograft nude mice with human gastric cancer cells and its molecular mechanism. Methods:Human gastric cancer cells were planted into nude mice in order to establish the cancer model, the different dosages of EGCG were injected intraperitoneally in the nude mice. After treatment, flow cytometry (FCM) was used to detect the apoptosis of implanted tumor cells. Immunohistochemical staining was used to detect the expression of apoptosis-related genes like Bal-2 and Bax in implanted tumor.Results:EGCG significantly inhibited tumor growth after being injecting intraperitoneally in the nude mice. The apoptotic cells in implanted tumor could be detected by flow cytometry with PI staining. The expressions of Bax、Caspase-3 were upregulated and Bcl-2 expression was downregulated in implanted tumor.Conclusions:EGCG could significantly inhibit tumor growth in xenograft nude mice with human gastric cancer cells through inducing apoptosis. The down-regulation of Bcl-2 expression and up-regulation of Bax expression observed could result in the activation of Caspase-3, the pathway might account for the induction of apoptosis.
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Objective:To construct a human Fab fragment phage display library and provide a platform for human antibody preparation.Methods:Peripheral blood lymphocytes were collected from healthy donor.The heavy chain Fd fragment and light chain of human immunoglobulin's genes were amplified by RT-PCR,and then cloned into phagemid pComb3XSS to generate human phage antibody library.Cutting with endonucleases such as SacⅠ,XbaⅠ,XhoⅠand SpeⅠto identify the insertion of the light chain or heavy chain Fd genes.IL-2 and digoxin as the antigen was used to scan the phage antibody library.Results:A phage antibody library of Fab had 8.4?107 members and it's recombinant rate was 70%.Through DNA sequencing of one positive clone,it was showed that its heavy chain belonged to IgG subvariety and its light chain to ? family.Conclusion:The success of constructing a nave human phage antibody library proves the useful of phage display system in human antibody preparation,and it can be used to select,purify and express of amylin Fab antibody.
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Aim To investigate the arrest effect of diallyl disulfide(DADS) in the cell cycle of human nasopharyngeal carcinoma SW480 cell line and its molecular mechanism.Mothdes The growth inhibition effect of DADS of different concentrations on SW480 cell line was measured by MTT assay and cell counting.Phase distribution of cell cycle was analyzed by flow cytometry.Expression of Ubiquitin and FKBP was determined by Western blot.Results The MTT assay showed that DADS inhibited growth of SW480 cells significantly in a dose-dependent manner.Adding 25,35,50 and 70 mg?L-1 DADS for 48 hours,SW480 cell growth was suppressed by 18.67%,33.02%,49.12% and 66.86%,respectively.There were significant differences between the treated and controlled cases(P
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Objective To prepare human Fab fragment antibodies against Interleukin-2 and identify their antigenic specificity and combining activities with antigens. Methods The specific Anti-interleukin-2 clones were screened from a natural human Fab fragment antibodies phage display library against the immobilized interleukin-2 antigens. Then the phagemid DNA from the specific clones was digested with Spe I and Nhe I to delete gⅢ (about 660bp). The digested 4.7kb DNA, which was purified after separation of bands from agarose gel using purification kit, was ligated with T4-DNA ligase and the ligated reaction mixture were transformed to the BL21 (DE3) pLysS. Positive clones on the LB agar plates were inoculated to liquid LB culture medium, and when the bacteria were grown to OD600≈0.5 at 37℃ with continuous shaking, IPTG was added to induce the expression of soluble Fab fragment antibodies at 30℃. The expressed products containing Fab fragment antibodies were determined by SDS-PAGE, Western blot and ELISA. Results The soluble products were identified as containing human Fab fragment antibodies against Interleukin-2 by Western blot and formed a Mr 47?103 band under non-reducing condition on SDS-PAGE. The band was then proved as anti-human Fab fragment antibodies by Western blotting. ELISA demonstrated that Fab fragments possessed good antigenic specificity as well as excellent combining activity with interleukin-2 antigens, and the fragments did not react with bovine serum albumin and IL-4 in ELISA. Conclusions The soluble human anti-interleukin-2 Fab fragment antibodies have been highly expressed and successfully identified, and an effective way has been searched out for constructing the engineering antibodies. All of the results may lay a potentially good foundation for engineering human Fab antibodies, and for the clinical application of the antibodies on the immunotherapy of tumor diseases.
