Objective To evaluate the performance of enzyme linked immunosorbent assay (ELISA) kit in detection of Hepatitis B virus(HBV) markers by using improved and optimized method ,so as to provide a practical and feasible method and reagents for clinical laboratory .Methods ELISA test was used for the detection of HBV markers .The gradient dilution method was used to e‐valuate the lower limit .The verifiation of cut off value was carried out based on clinical and laboratory standards institute (CLSI) EP12‐A2 document .Samples with cut off values were collected to evaluate the precision ,including repeatability and intermediate precision .The coincidence rates were counted through comparing the results of ELISA with those of external quality assessment and those detected by using Abbott i4000SR chemiluminescence instrument .Results The lower detection limit of HBsAg ,HBsAb , HBeAg ,HBeAb and HBcAb were 0 .2 IU/mL ,20 mIU/mL ,1 NCU/mL ,0 .75 NCU/mL and 0 .05 NCU/mL respectively .The cut‐off value(C50 )± 20% concentration included the concentration range between C5 and C95 .The within‐run coefficient of variation (CV)≤15% ,in sandwich method the between‐run CV≤25% ,in competition method the between‐run CV≤35% .The positive and negative coincidence rates in accuracy and comparing with i 4000SR all were more than 95% and all κ>0 .75 .Conclusion ELISA tests for HBV markers in our laboratory could meet the requirements of the detection performance and clinical needs .

Objective To observe the effects of tissue factor pathway inhibitor(TFPI) on thrombosis formation in rabbit carotid arteries after ballon injury. Methods Fouty rabbits with the weight 2.5-3.0 kg were respectively divided into 4 groups, Ad-TFPI, Ad-LacZ, PBS and normal control groups. The normal control group was not given any treatment and other 3 groups were given 0.2 ml Ad-TFPI, Ad-LacZ or PBS reproduced by the Dispatch catheter respectively after the PTCA balloon iniury on the right carotid arteries. Ten days after gene transfer the repeated balloon injury was performed in the 3 groups, and the first balloon injury was performed in the normal control group by the same method. The carotid blood flow was recovered immediately after the injury. Thirty minutes later all the animals were sacrificed. The injured carotid arteries and one part of contralateral normal artery were cut down, scissored along the long axis, flattened and fixed in the 2% glutaral. The platelet aggregation and thrombosis formation on the luminal surfaces was observed under electron microscope. Results The electron microscope results showed that the vascular endothelial cell structure was integrated and lined up in order in the nomal artery which had no any injury. After the balloon injury in the normal control group, the structure of the endothelial cell was disintegrated, and there was some platelet aggregation but no fibrosis formation. A large amount of platelet aggregated but no fibrosis formed in Ad-TFPI group after the repeated balloon injury. A large amount of fibrosis formed and red cells piled up in the Ad-LacZ and PBS group. The positive rate of thrombosis formation among groups had siginificant differences(χ2=14.95, P<0.01). The positive rate in Ad-TFPI group(20%) was lower than that in Ad-LacZ group(80%, χ2=7.20, P<0.01) and PBS group(70%, χ2=5.05, P<0.05), but was higher than that in the normal control group(10%, χ2=0.39, P>0.05). The positive rate in Ad-LacZ group(80%) was higher than in the normal control group(10%, χ2=9.90, P<0.01) and in the PBS group(70%, χ2=0.27, P> 0.05). The positive rate in PBS group(70%) was higher than that in the normal control group(10%, χ2=7.50, P< 0.01). Conclusions The repeated balloon injury method can cause a large amount of fibrosis formation in the rabbit carotid. TFPI gene inhibits thrombosis formation in balloon-injured rabbit carotid arteries.

OBJECTIVETo analyze CD21 expression in diffuse large B cell lymphoma (DLBCL) and to explore its relationship with the clinicopathological characteristics and prognosis.

METHODSThe clinical data from 80 DLBCL patients who were treated in First Hospital of Jilin University from June 2005 to September 2011 were retrospectively analyzed. The cases were subjected to immunohistochemical staining (SP method) for Ki-67, CD20, CD79a, CD3, CD43, CD5, cyclin D1, bcl-2, CD10, bcl-6, GCET-1, FOXP-1 and MUM-1 protein expression in the tumor tissue. Immunohistochemistry was also used to detect CD21 expression in the tumor tissue. SPSS 18.0 was used to analyze the relationship between CD21 expression and various clinical factors, and the relationship between various clinical factors including CD21 and overall survival.

RESULTSIn the patients aged under 60 years, the incidence of CD21(+) lymphoma (64.0%, 16/25) was significantly higher than that of CD21(-) lymphoma (38.2%, 21/55). There were more CD21(+) lymphoma patients who were at clinical stages I-II (52.0%, 13/25) than patients with CD21(-)lymphomas (23.6%, 13/55). There were also more CD21(+) lymphoma patients (68.0%, 17/25) having less than two extranodal sites involvement than CD21(-)lymphoma patients (41.8%, 23/55). In addition, there were more CD21(+) lymphoma patients with IPI 0-2 (68.0%, 17/25) than CD21(-)lymphoma patients (41.8%, 23/55). There were more CD21(+) lymphoma patients with GCB subtype (60.0%, 15/25) than CD21(-)lymphoma patients (23.6%, 13/55). Death related to DLBCL was less in CD21(+) lymphoma patients (32.0%, 8/25) than CD21(-) lymphoma patients (56.4%, 31/55). Univariate analysis showed that these clinical pathological characteristics affected the overall survival of DLBCL patients, including age, ECOG score, LDH, extranodal involvement, IPI index, CD21 expression, treatment option and efficacy (P < 0.05) . Cox multivariate analysis showed that ECOG score, LDH, extranodal involvement, CD21 expression were closely related to prognosis, and the difference was statistically significant (P < 0.05). Among the 80 patients, the overall survival (OS) of CD21(+) lymphoma patients was significantly higher than that of CD21(-) lymphoma patients.

CONCLUSIONSThe expression of CD21 is associated with young age at onset, early clinical stage, small number of involvement and low IPI index. The OS and median overall survival of CD21(+) lymphoma patients are significantly higher than those of CD21(-) patients. CD21 expression, ECOG score, LDH, extranodal involvement are independent prognostic factors in DLBCL, and in particular, the expression of CD21 is more significant in the prognosis of DLBCL patients.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Child ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Female ; Gastrointestinal Neoplasms ; pathology ; Germinal Center ; pathology ; Humans ; Immunohistochemistry ; L-Lactate Dehydrogenase ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Prednisone ; therapeutic use ; Prognosis ; Receptors, Complement 3d ; metabolism ; Retrospective Studies ; Survival Rate ; Vincristine ; therapeutic use ; Young Adult

OBJECTIVETo investigate the in vivo gene expression of adenovirus-mediated human tissue factor pathway inhibitor (hTFPI) and its inhibition effects on intimal proliferation in rabbit carotid arteries after balloon injury.

METHODSRabbits underwent carotid artery balloon injuries were treated with Ad-TFPI (n = 25), Ad-LacZ (n = 25) or PBS (n = 10), respectively. Sham operated rabbits (n = 10) serve as normal controls. The expressions of human TFPI at mRNA and protein levels were detected by RT-PCR and ELISA respectively on the 3rd, 7th, 10th, 14th, 28th day after operation. Intimal proliferation was detected by angiograms and morphometric analysis.

RESULTSTFPI mRNA and protein expressions were detected at 3 days and peaked at the 10th and 14th day after TFPI gene transfer. The expressions were still detectable on the 28th day. There was no TFPI expression in Ad-LacZ group. The carotid angiogram results indicated that the minimal lumen diameter in TFPI group was significantly larger and the lumina stenosis percentage was significantly lower in TFPI group compared those in Ad-LacZ and PBS groups (all P < 0.05). The morphometric analysis showed that the intimal area, the ratio of the intimal/media area, the lumina stenosis percentage in TFPI group were all significantly reduced compared with those in Ad-LacZ and PBS groups (all P < 0.01).

CONCLUSIONSThe TFPI gene could be effectively transferred by adenovirus vector to injured carotid arteries and transferred Ad-TFPI could significantly attenuate intimal proliferation in balloon injured carotid arteries in rabbits.

Adenoviridae ; genetics ; Animals ; Carotid Arteries ; metabolism ; Carotid Artery Injuries ; metabolism ; pathology ; Female ; Gene Transfer Techniques ; Genetic Vectors ; Humans ; Lipoproteins ; genetics ; Male ; Rabbits ; Transfection ; Tunica Intima ; pathology

BACKGROUNDPulmonary embolism (PE) can be difficult to diagnose in elderly patients because of the coexistent diseases and the combination of drugs that they have taken. We aimed to compare the clinical diagnostic values of the Wells score, the revised Geneva score and each of them combined with D-dimer for suspected PE in elderly patients.

METHODSThree hundred and thirty-six patients who were admitted for suspected PE were enrolled retrospectively and divided into two groups based on age (≥65 or <65 years old). The Wells and revised Geneva scores were applied to evaluate the clinical probability of PE, and the positive predictive values of both scores were calculated using computed tomography pulmonary arteriography as a gold standard; overall accuracy was evaluated by the area under the curve (AUC) of receiver operator characteristic curve; the negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were calculated.

RESULTSNinety-six cases (28.6%) were definitely diagnosed as PE among 336 cases, among them 56 cases (58.3%) were ≥65 years old. The positive predictive values of Wells and revised Geneva scores were 65.8% and 32.4%, respectively (P < 0.05) in the elderly patients; the AUC for the Wells score and the revised Geneva score in elderly was 0.682 (95% confidence interval [CI]: 0.612-0.746) and 0.655 (95% CI: 0.584-0.722), respectively (P = 0.389). The negative predictive values of D-dimer, the Wells score combined with D-dimer, and the revised Geneva score combined with D-dimer were 93.7%, 100%, and 100% in the elderly, respectively.

CONCLUSIONSThe diagnostic value of the Wells score was higher than the revised Geneva score for the elderly cases with suspected PE. The combination of either the Wells score or the revised Geneva score with a normal D-dimer concentration is a safe strategy to rule out PE.

Aged ; Aged, 80 and over ; Angiography ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Male ; Pulmonary Embolism ; diagnosis ; diagnostic imaging ; metabolism ; Retrospective Studies

OBJECTIVETo establish a rat model of heatstroke complicated by endotoxemia for studying the pathogenesis of severe heatstroke.

METHODSMale specific pathogen-free Wistar rats were randomly assigned into 4 groups, namely normothermic saline group (group C), heat exposure saline group (group H), normothermic LPS group (group L), and heat exposure LPS group (group HL). The rectal temperature (Tr), heart rate (HR), mean arterial pressure (MAP), and respiratory rate (RR) of the rats receiving different treatments were continually monitored and their white blood cell count (WBC) and histology of the lungs were observed at 0, 40, 80 and 120 min after the treatments.

RESULTSThe rats in HL-Group displayed significantly higher Tr (43.04+/-0.11 degrees C), HR (660+/-42 beats/min), and RR (150+/-11/min) but lower MAP (49.0+/-3.5 mmHg) as compared with the C Group. There were significant differences in the values of Tr, HR, RR and MAP between HL and group L and in HR and MAP between H groups HL and. The rats in group H displayed significantly higher WBC than group C. In contrast, the rats in L groups HL and had significantly lower WBC. LPS injection and heat stress induced pulmonary edema and features characteristic of acute microvascular lung injury in the rats.

CONCLUSIONThe rat model established by LPS injection and heat stress can successfully mimic the development of severe heatstroke after LPS challenge and heat stress, and provides a suitable model for studying the primordial role of the lungs in the pathogenesis of severe heatstroke.

Animals ; Disease Models, Animal ; Endotoxemia ; chemically induced ; complications ; Heat Stroke ; complications ; Lipopolysaccharides ; Male ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVEThe ratio of psychological to organic ED changes with aging. This study aimed to analyze the results of nocturnal electrobioimpedance volumetric assessment (NEVA) for ED patients of different age groups and their significance in the diagnosis of ED.

METHODSA total of 83 ED patients were divided into 4 age groups (< or = 29 yr, 30 -39 yr, 40 -49 yr and > or = 50 yr) and detected for nocturnal penile tumescence (NPT) by NEVA.

RESULTSThirty-four of the cases were diagnosed as organic ED, and the other 49 as psychological ED. With the increase of age, the former was increased from 30.3% in the < or = 29 yr group to 60.0% in the > or = 50 yr group, while the latter decreased from 69.7% to 40.0%.

CONCLUSIONThe percentage of organic ED tends to grow with the increase of age, while that of psychological ED is just the opposite.

Adult ; Aging ; Electric Impedance ; Erectile Dysfunction ; diagnosis ; physiopathology ; Humans ; Male ; Middle Aged ; Penile Erection

OBJECTIVETo analyse the efficacy and safety of thalidomide (Thal) for patients with multiple myeloma (MM).

METHODSEffectiveness and adverse events of 102 MM patients treated with thalidomide at a median dosage of 200 mg/d. Thirteen cases were treated with Thal alone (group A), and 105 case with Thal in combination with other therapeutic agents (group B) were retrospectively analyzed.

RESULT1) The response rate (RR) (CR + PR) was 65.4% for induction therapy in 52 cases and 45.5% for salvage therapy in 66 cases. RR in group B was higher than that in group A (58.1% versus 23.1/%, P= 0.017), and the non-response/progress (NR) rate was lower (15.2% versus 46.2%, P= 0.015). In group B, the NR rate was lower in 50 cases of newly diagnosed MM than in 55 cases of refractory or relapsed MM (6.0% versus 23.6%, P=0.012). In group B, RR between Thal+VAD or M, regimen (72 cases) and Thal + MP regimen (33 cases) was not statistically significant (62.5% versus 48.5%, P >0.05). 2) The median duration of response maintenance was 15.5 (1.0-58.0) months in 21 cases. 3) Among 97 patients with follow-up data, the estimated median duration of OS was 44 months in a median follow-up duration of 20 months and the accumulative time for use of Thal was 8 months. In univariate analysis,the accumulative duration for use of Thal 6 months, hemoglobin > or = 100 g/L and bone marrow megakaryocytes > 20 per smear were associated with longer OS (P = 0.0014, 0.0101, 0.019, respectively). 4) Multivariate analysis suggested that the accumulative time for use of Thal and bone marrow megakaryocytes > 20 were independent good prognostic factors for OS (P = 0.006, 0.036, respectively). 5) The adverse events of Thal were mostly endurable, the rate of thrombus events was lower than that reported in literature.

CONCLUSIONThalidomide alone or combined with chemotherapy is an useful therapy for MM. The accumulative time for use longer than 6 months may improve survival.

Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; Prognosis ; Retrospective Studies ; Thalidomide ; adverse effects ; therapeutic use ; Treatment Outcome

OBJECTIVETo investigate the changes in the activity of Escherichia coli asparaginase (L-asp) and the concentration of asparagines (ASN) in the plasma of the acute lymphoblastic leukemia (ALL) children receiving L-asp containing chemotherapeutic protocol to explore more reasonable usage of L-asp in the treatment of childhood ALL.

METHODSL-asp containing hemotherapy regimen of VDLP was used, in which L-asp (10,000 U/m(2)) was administered intravenously every other day for 10 doses in 15 children with ALL. A total of 340 peripheral blood samples were collected at scheduled time points during the therapy and plasma L-asp activity (by spectrophotometric assay) and asparagines concentration (by RP-HPLC) were measured.

RESULTSDuring the administration of L-asp, the plasma L-asp activity was increasing gradually peaked after eight doses and then decreased gradually, while the plasma concentration of asparagines maintained in complete or nearly complete depletion status. After the therapy courses finished, a plasma L-asp activity above 100 U/L with asparagines almost complete depletion status was lasting for about seven days.

CONCLUSIONThe current L-asp containing chemotherapeutic protocols in which L-asp was administered in a dose of 10 000/m(2) intravenously every other day, are efficient enough for the depletion of plasma ASN.

Adolescent ; Antineoplastic Combined Chemotherapy Protocols ; blood ; pharmacokinetics ; therapeutic use ; Asparaginase ; administration & dosage ; blood ; pharmacokinetics ; Asparagine ; blood ; Child ; Child, Preschool ; Drug Administration Schedule ; Female ; Humans ; Infusions, Intravenous ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; blood ; drug therapy ; Treatment Outcome