[Changing characteristic of blood coagulation factors and their correlation with blood coagulation status in different hepatic diseases].

OBJECTIVE: To investigate the correlation between pro coagulation factors and anti-coagulation factors synthesized by the liver, and the correlation between fibrin degradation products (FDP) and D-dimer (D-D) concentration and coagulation proteins synthesized by extra-hepatic tissues, in different liver diseases; to explore the relationship between coagulation and bleeding in hepatic diseases. METHODS: Chronic hepatitis B (CHB) patients, CHB-related liver cirrhosis patients, CHB-related liver failure patients and healthy (normal) controls were selected for study and provided blood samples for analysis. The activity of coagulation factors (F) II, V, VII, VIII, IX, X, XI, and XII was detected using the one-stage clotting method. Coagulogram analysis, including activated partial thromboplastin time (APTT), thrombin time (TT), and prothrombin time (PT), was conducted by the solidification method. Antithrombin III (AT-III) and protein C (PC) activities were measured by chromogenic substrate assay. FDP concentration was detected using immunoturbidimetry. Tissue factor pathway inhibitor (TFPI), thrombomodulin (TM), von Willebrand factor (vWF), and tissue factor (TF) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: With the exception of FVIII, coagulation factors and anticoagulant proteins synthesized by the liver were decreased and the coagulogram was extended for all patients. Likewise, the FDP and D-D concentrations were increased in blood. CHB patients, however, presented with increased levels of FVIII, TFPI, TM, vWF, and TF. Pairwise comparison indicated statistical differences existed among CHB, CHB-related liver cirrhosis, and liver failure patients: TFPI: 239.3+/-206.4, 315.0+/-258.6, and 319.5+/-298.1 -- higher than normal control: 104.0+/-87.1, F = 5.453, P less than 0.05; vWF: 70.3+/-29.5, 105.5+/-58.0, and 179.3+/-61.7 -- higher than normal control: 21.9+/-7.2, F = 20.104, P less than 0.05; TF: 85.9+/-85.7, 234.2+/-202.9, and 344.7+/-214.6 -- higher than normal control: 12.8+/-8.1, F = 8.619, P less than 0.05; FVIII: 157.2+/-53.4, 206.9+/-86.9, and 335.7+/-117.7 -- higher than normal control: 105.5+/-46.2, F = 13.418, P less than 0.05. CONCLUSION: In parallel to the progression of liver diseases, pro coagulation and anti-coagulation elements synthesized by the liver were reduced. In contrast, fibrinolysis activity was enhanced, which is expected to lead to an imbalance between blood clotting and anti-clotting factors. This may be an important cause for the bleeding that occurs in end-stage liver disease. Expressions of TFPI, TM, vWF, and TF significantly change in the early stage of liver diseases, as compared to normal (healthy) levels, and may represent a sensitive indicator of vascular injury.

[Effects of extracellular calcium concentration on platelets aggregation, coagulation indices and thromboelastography].

OBJECTIVE: To detect the effect of extracellular Ca2+ concentrations on test results of coagulation-related parameters. METHODS: Blood samples of outpatient medical volunteers were collected and then different doses of calcium chloride added. The rate of platelet aggregation (n = 42), prothrombin time (PT), thrombin time (TT) and activated partial thromboplastin time (APTT) (n = 21) and parameters of thromboelastography (n = 30) were detected according to the standard protocols by plasma turbidimetry, coagulation and recalcification respectively. RESULTS: When the plasma Ca2+ concentration was in the range of 0.1 - 33.7 mmol/L, the rate of platelet aggregation gradually increased with a increasing concentration of Ca2+. And the rates induced by adenosine diphosphate (ADP) and arachidonic acid (AA) were (51.8 +/- 9.6)% - (94.7 +/- 4.8)% and (64.4 +/- 12.2)% - (93.2 +/- 5.5)% respectively. When the Ca2+ concentration was 39.0 mmol/L, the rate decreased markedly [ADP (9.1 +/- 5.3)%, AA (11.1 +/- 4.5)%, both P < 0.01]. When the Ca2+ concentration was in the range of 0.1 - 33.7 mmol/L, the values of PT gradually increased with a increasing concentration of Ca2+. The values of TT changed in "V"-type and became minimum when the calcium concentration was 4.4 mmol/L. The values of APTT decreased with higher calcium concentrations and could not be determined when the concentration increased above 0.5 mmol/L. When the Ca2+ concentration was in the range of 0.4 - 27.3 mmol/L, the values of reaction time and coagulation time of thromboelastography changed in "V"-type and became nearly minimal at the Ca2+ concentration of about 2.1 mmol/L. The values of alpha angle and maximum amplitude changed in "V"-type and became maximal at the Ca2+ concentration of 2.1 mmol/L. CONCLUSIONS: The effect of Ca2+ concentration on the testing results of coagulation-related parameters is significant. A high calcium ( > or = 39 mmol/L) can inhibit the platelet aggregation, coagulation factor activity and blood coagulation. The Ca2+ concentration of 2.1 mmol/L seems to be the optimal concentration for thromboelastography by recalcification method.

[Identification of differential proteins in the seminal plasma of healthy fertile and non-obstructive azoospermia men by shotgun proteomic strategy].

OBJECTIVE: To identify differential proteins in the seminal plasma of healthy fertile men and non-obstructive azoospermia patients by the shotgun proteomic strategy. METHODS: Six seminal plasma samples from 3 healthy fertile and 3 non-obstructive azoospermia volunteers were collected by Percoll isolation, balanced-mixed, and followed by separation of the mixture by SDS-PAGE. The proteins were subjected to in-gel enzymolysis and isolation of peptide fragments, and then identified by the shotgun proteomic strategy. Then comparative analyses were made between the two groups on the identified proteins with the unique peptide count > or = 2 and = 1 but with the peptide count > or = 4. RESULTS: A total of 213 differential proteins were identified, 133 in the non-obstructive azoospermia patients and 80 in the healthy fertile men. According to the molecular function, these differential proteins mainly fell into the types of signal transduction, cytoskeleton and catalytic activity, especially oxidoreductase activity in the latter type. Eighteen of the differential proteins were found to be of particular significance, including dynein heavy chain, fatty acid synthase, and tubulin alpha-6 chain. CONCLUSION: The differential proteins identified in this study were many in number and various in function, which not only demonstrated the value of the shotgun proteomic strategy in protein identification, but also suggested the complicated pathogenesis and varied types of non-obstructive azoospermia. The samples must be selected strictly based on their gene and histological types. Non-obstructive azoospermia was shown to be related with the M phase of the mitotic cell cycle at the protein level, but its specific mechanism remains unknown.

[Identification of asthenozoospermia-associated proteins in human seminal plasma by shotgun proteomic strategy].

OBJECTIVE: To identify asthenozoospermia-associated proteins in seminal plasma by the shotgun proteomic strategy. METHODS: Six seminal plasma samples were collected by Percoll respectively from healthy fertile and asthenozoospermia volunteers, balanced, mixed, and then the mixture was separated by SDS-PAGE. The proteins in the gel were enzymolyzed, extracted and identified by the shotgun proteomic strategy. The identified proteins with the unique peptide count > or =2 or the unique peptide count=1 but the total count > or =4 were compared between the two groups. RESULTS: A total of 172 differential proteins were identified, of which, 89 were exclusively from the asthenozoospermia and 83 exclusively from the healthy fertile men. According to the molecular function, these differential proteins were mainly the types of signal transduction and catalytic activity. CONCLUSION: Functionally, 10 of the proteins are particularly important, which include annexin VI isoform 2, isoform 1 of interleukin-6 receptor subunit beta precursor, Mr 400,000 protein, cytosolic dynein heavy chain, alpha-actinin-4, receptor-type tyrosine-protein phosphatase eta precursor, vitamin D-binding protein precursor, protein S100-A11, protein S100-A9 and ANXA4.

[Identification of proteins in the seminal plasma of healthy fertile men by shotgun proteomic strategy].

OBJECTIVE: To identify proteins in the seminal plasma of healthy fertile men. METHODS: Three seminal plasma samples were collected from healthy fertile volunteers by Percoll isolation, and then the balanced mixture of the seminal plasma was separated by SDS-PAGE. The proteins in the gel band underwent enzymoloysis, and was extracted and identified by shotgun proteomic strategy. RESULTS: A total of 331 proteins were identified, with the molecular weight (MW) ranging from 8 000 to 572 068 and the isoelectric point (pI) from 4.36 to 11.05. Based on the molecular function and biological process of the proteins, 51 (15.4%) were classified as transport proteins, 11 (3.32%) as cell movement proteins, 63 (19.03%) as signal transduction proteins, 147 (44.4%) as proteases, 38 (11.5%) as enzyme regulator proteins, 21 (6.3%) as programmed cell death proteins, 12 (3.62%) as structural proteins and 59 (17.8%) as proteins with unknown molecular function. CONCLUSION: Shotgun proteomic strategy is a good method for protein identification. Annexin A, Annexin-associated proteins and the Ras-related protein Rab were the major members of the signal transducer proteins identified. Ca2+ and G protein signal pathways may play a most important role in the extracellular signal transduction into cells, but the interactions between these proteins remain unknown. The great quantity of enzymes and enzyme regulator proteins identified in the seminal plasma may be closely related with the maintenance of sperm motility and metabolism.

[Protein compositions in the seminal plasma are similar between severe oligospermic and healthy fertile men].

OBJECTIVE: To analyse the variability of proteins in the seminal plasma of severe oligospermic and healthy fertile men. METHODS: Spermatic fluid samples were collected from 11 healthy fertile men and 6 severe oligospermic male volunteers and tested by SELDI-TOF-MS with the CM10 protein chip to get the protein spectra maps, which were automatically treated with the special softwares of Ciphergen Inc. RESULTS: The mean peak heights of 2 lower-abundance proteins expressed in the seminal plasma of the severe oligospermic men were statistically different from the healthy fertile males (P<0.05). Fifteen different proteins existed between the nonobstructive azoospermic and the severe oligospermic group, 7 of which, with m/z of 7,196.058, 7,547.610, 5,780.493, 7,059.844, 7,409.589, 5,379.173 and 10,778.810, also between the non-obstructive azoospermic and the healthy fertile males (P<0.05). Except the latter two, the contents of the other 5 proteins were decreased in the non-obstructive azoospermic men (P<0.05). CONCLUSION: The finger prints of the seminal plasma proteins of the severe oligospermic group were similar to those of the healthy fertile males, both significantly different from the non-obstructive azoospermic men. It is suggested that pathogenesis mechanisms differ exist between non-obstructive azoospermia and severe oligospermia but are not the simple accumulation of genetic factors.

[Difference in examination of coagulation function in heparin anticoagulant therapy between point of care tests and automatic coagulation analyzer].

OBJECTIVE: To investigate the difference in coagulation screening in heparin anticoagulant therapy between dry-chemistry point of care tests (POCT) and automatic analyzer. METHODS: Blood samples were collected from 44 patients with myocardial infarction or intracranial hemorrhage, 34 males aged 78 +/- 10 and 10 females aged 73 +/- 8, to undergo examination of prothrombin time (PT), international normalized ratio (INR), activated partial thromboplastin time (APTT), and fibrinogen (Fbg) by dry-chemistry methods with coagulation analyzers of the models CG02 and Hemochron Jr II. The levels of APTT of 30 patients were measured by the bedside with analyzers model Hemochron Jr II, after their anticoagulation samples underwent measurement of the plasma heparin activity with ACL Advance automatic coagulation analyzer; the APTT was determined with STA-R. RESULTS: The correlation coefficients of PT, INR, APTT, and Fbg between the CG02 and STA-R results were 0.945, 0.966, 0.662, and 0.977 respectively (all P < 0.05). However, t test showed that the levels of PT and INR between these 2 groups were not significant, and the differences in the levels of APTT and Fbg were significant (both P < 0.05). ACL Advance analyzer showed that the heparin activities of 30 samples were 172 approximately 1465 U/L, and the APTT values measured by Hemochron Jr II and STA-R were 87 s +/- 32 s and 90 s +/- 46 s (P < 0.01). CONCLUSION: Dry-chemistry POCT is not suitable to report the results of the same patient simultaneously with the automatic analyzer.

[Comparative analysis of proteins in seminal plasma of non-obstructive azoospermia patients and healthy fertile males].

OBJECTIVE: To analyse protein alterations in the seminal plasma of non-obstructive azoospermia patients. METHODS: Semen samples were collected from 11 healthy fertile and 6 azoospermia male volunteers respectively and tested by SELDI-TOF-MS with CM10 protein chip to get protein spectra maps, which were automatically treated with the special softwares of Ciphergen Inc. RESULTS: The mean peak heights of 28 proteins expressed in the seminal plasma of the azoospermia patients were statistically different from those of the healthy fertile males (P < 0.05 ), of which 24 were of lower contents than in the normal controls, 4 with remarkably significant difference, M/Z 7 196.058, 7 630.573, 7 547.610 and 7 709.833 (P < 0.01). CONCLUSION: The seminal plasma proteins of the azoospermia patients were significantly different from those of the healthy fertile males, with decreased contents of most of the different proteins, which might be significantly correlated with the development of azoospermia.

[Study of two electrophoresis procedures of seminal plasma proteins].

OBJECTIVE: To analyse clinical feasibility of two electrophoresis procedures of seminal plasma proteins, agarose gel electrophoresis and SDS-agarose gel electrophoresis. METHODS: Sixty-nine semen samples were examined and classified into three groups: the asthenozoospermia (n = 22), the asthenoteratozoospermia (n = 19), and the relative normal group (n = 28) with normal routine and special test results, according to WHO routine and special test criterion. Then, the seminal plasma protenis were separated by two different electrophoresis, with SDS-agarose and agarose support medium, the buffer pH 7.0 and 9.2 respectively. The agarose gel electrophoresis was done under various sample loading time, motion power and staining modules. The completed gels were scanned and compared the each other statistically. RESULTS: Seminal plasma proteins can be separated into 4 strips by SDS-agarose gel electrophoresis with acid crystal violet, and the strips were diffusion and with dark background. However, 6 clear strips named A, B, C, D, E, and F can be obtained by agarose gel electrophoresis with 6 min. After samples were loaded and stained by amidoblack, there showed appropriate spaces among strips, and it was very easy to scan the drying gel by a densitometer. Using agarose gel electrophoresis, the statistical difference in strip C and E was shown between the asthenozoospermia and the relative normal group, and between the asthenozoospermia and the asthenoteratozoospermia, however, not between the relative normal and the asthenoteratozoospermia group. Moreover, the samples in the relative normal group with normal routine and special test results were in different electrophoresis maps. CONCLUSION: Agarose gel electrophoresis of seminal plasma proteins with buffer pH 9.2, 6 min. sample loading and amidoblack stain was a simple, fast and fit technique for clinic.

[Evaluation of PFA P2Y assay in monitoring platelet function in elderly patients with cardiovascular disease receiving clopidogrel treatment].

OBJECTIVE: To evaluate the value of a new platelet function test PFA P2Y (PFA-200) in monitoring clopidogrel treatment for cardiovascular disease in elderly patients. METHODS: Fifty-six elderly patients receiving clopidogrel therapy in the Department of Cardiology of General Hospital of PLA from March to August in 2016 and 85 healthy volunteers were recruited for analysis. All the subjects underwent PFA P2Y, LTA (light transmittance aggregometry) and TEG (Thromboelastograph) tests, and Spearman correlation coefficients were used to test the associations between test results. The agreement among the 3 platelet function test methods was assessed using Cohen's kappa coefficient. RESULTS: Correlation coefficient (r) was -0.701 (P<0.001) between PFA P2Y and LTA, and 0.475 (P<0.001) between PFA P2Y and TEG. The agreement was 75% between PFA P2Y and LTA and 67.9% between PFA P2Y and TEG. The κ value was 0.434 (P=0.001) between PFA P2Y and LTA and 0.242 (P=0.046) between PFA P2Y and TEG. With ADP-induced maximum platelet aggregation rate of LTA >50% as the laboratory clopidogrel resistance, the cut-off value of PFA P2Y was 119 s (AUC=0.733) with a sensitivity of 75.6% and a specificity of 73.3%. CONCLUSION: PFA P2Y has a moderate correlation and agreement with LTA, but has a poor correlation and agreement with TEG. PFA P2Y can be useful for assessing the effects of clopidogrel therapy and the association of the cut-off value (119 s) with the long-term clinical ischemic events needs be confirmed in further study.

[The value of coagulation factors in severity categorization of hepatitis B-related liver cirrhosis].

OBJECTIVE: To investigate the diagnostic value of coagulation factors in assessing the severity degree of liver cirrhosis caused by hepatitis B. METHODS: Fifty-eight patients with liver cirrhosis and twenty healthy persons as control were enrolled. Prothrombin time activity percentage (PTA), activated partial thromboplastin time, coagulation activity of factor II, V, VII, VIII, IX and X were detected with clotting assay. Antithrombin-III (AT-III) was detected with colorimetric assay. The biochemical markers were also detected. RESULTS: The differences of PTA, factor II, VII and AT-III among Child-Pugh A, B, C in patients with liver cirrhosis were statistically significant (P < 0.01). Through receiver operating characteristic curve analysis, when 64% and 50% were used as cut-off values for PTA and factor VII in diagnosing Child-Pugh B, the area under the curve (AUC) was 0.689 and 0.610, the sensitivity was 76.9% and 61.5%, the specificity was 62.2% and 55.6%; when 54% and 39% were used as cut-off values for PTA and factor VII in diagnosing Child-Pugh C, the AUC was 0.924 and 0.942, the sensitivity was 80.0% and 86.7%, the specificity was 88.4% and 90.7%. Stepwise linear regression was done between Child-Pugh grade and coagulation factors. PTA, cholinesterase (Che), total bilirubin (TBil), albumin (Alb), factor VII were included in regression equation, Y = 15.008 - 0.018 x PTA - 0.288 x Che + 0.264 x TBil - 0.988 x Alb - 0.034 x VII, R(2) = 0.871. Patients whose Y was less than 8 were classified as grade "a", between 8 - 10 as grade "b", more than 10 as grade "c", the diagnostic accuracy was 84.5%. CONCLUSION: Coagulation factor VII may serve as a helpful marker in diagnosing the severity degree of liver cirrhosis.

[The relationship between hemostatic changes in liver cirrhosis patients with different degrees of liver lesions in reference to Child-Pugh scores].

OBJECTIVE: To investigate the relationship between hemostatic changes in liver cirrhosis patients with different degrees of their liver lesions. METHODS: Forty-three patients (35 men, 8 women; age: 25 to 71 yr) with liver cirrhosis were divided into three subgroups (A, B, and C) on the basis of Child-Pugh classification. Among the patients, 13 were classified as Child-Pugh class A, 15 were class B, 15 were class C. 16 healthy individuals served as controls. A series of hemostatic tests and parameters including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (Fib), factors II, V, VII, VIII, IX, X, vWF assay, antithrombin-III (AT-III), protein C (PC), D-dimer, tissue plasminogen activator antigen (t-PA), plasminogen activator inhibitor activity (PAI) were performed on 43 patients and the 16 healthy controls. RESULTS: PT and APTT were progressively prolonged from A to B and then to C. In comparison to the controls there was a significant difference. Fibrinolytic activity and the activities of factors II, V, VII, IX, X were progressively decreased from A to B and then to C. In comparison to the controls there was a significant difference . AT-III and PC activity were progressively decreased from A to B and then to C. In comparison to the controls there was a significant difference. D-dimer and t-PA-antigen were progressively increased from A to B and then to C. In comparison to the controls there was significant difference. PAI activity did not display significant changes in the four groups. CONCLUSION: We found that there is a close relationship between the severity of cirrhosis and the hemostatic changes. Because the deterioration of the coagulation function and increasing fibrinolytic activity parallel the severity of liver cirrhosis, adequate treatment for cirrhotic bleeding should not only correct the coagulation defects, but also lower the increased fibrinolytic activity.

[A discussion of standardization for prothrombin time in patients with advanced liver diseases].

OBJECTIVE: To determine which expression mode of prothrombin time (PT) might achieve PT standardization in patients with advanced liver diseases. METHODS: PT was measured with six thromboplastins with different ISI values in 16 severe chronic hepatitis patients, 50 decompensated liver cirrhosis patients and 30 patients on oral anticoagulation therapy. The results were expressed in PT (second), PTA (%), PTR and INR. RESULTS: In chronic hepatitis patients, the means of the six group's PTAs ranged from 24% to 34%, while their upper limits ranged from 47% to 61%. The means of the INRs ranged from 2.55 to 5.13, while their upper limits ranged from 4.65 to 12.77. Through one-way ANOVA of repeated measures, PPTA (0.489) was > PINR (0.120). In patients with liver cirrhosis, the means of the PTA in six groups ranged from 50% to 59%, while their upper limits ranged from 82% to 90%. The means of the INR ranged from 1.40 to 1.80, while their upper limits ranged from 1.97 to 3.69. Through one-way ANOVA of repeated measures, PPTA (0.102) was > PINR (0.01). In patients on oral coagulation therapy, the means of PTA ranged from 26% to 37%, while their upper limits ranged from 39% to 49%. The means of INR ranged from 2.35 to 2.66, while their upper limits ranged from 3.16 to 4.26. Through one-way ANOVA of repeated measures, PPTA (0.01) was less than PINR (0.102). The correlation between the results detected by Neoplastine and by other reagents were analyzed. They correlated well with each other when PTA was used as the expression mode of PT in patients with advanced liver disease. But in patients on oral anticoagulation therapy, when only the INR was used as the expression mode of PT, the correlation was well with each other. CONCLUSION: The use of INR provides inadequate standardization. Only when the PT is expressed in PTA, then it may provide a standardization mode in patients with advanced liver diseases.

[Establishing the reference range of venous blood measured by automated haematology analyzer in Chinese adults].

OBJECTIVE: To establish the reference range of venous blood measured by automated hematology analyzer in China. METHODS: Different hematoanalyzers were used to examine the white blood cell (WBC), red blood cell (RBC), hemoglobin (Hb), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular volume (MCV), and hemotocrit (HCT) of venous blood from 1,749 healthy people, 927 males and 822 females, aged 18 - 60, in 14 cities (Harbin, Changchun, Beijing, Tianjin, Lanzhou, Xi'an, Nanjing, Suzhou, Chengdu, Wuhan, Chongqing, Fuzhou, Kunming, and Guangzhou) of China. The examination was completed within the period of one month under strict in-laboratory quality control. RESULTS: The range of WBC was (3.97 - 9.15) x 10(9)/L in males and (3.69 - 9.16) x 10(9)/L in females. The range of RBC was (4.09 - 5.74) x 10(12)/L in males and (3.68 - 5.13) x 10(12)/L in females. The range Hb was 131 - 172 (151) g/L in males and 113 - 151 (129) g/L in females. The range of MCH was 27.8 - 33.8 (30.8) pg in males and 26.9 - 33.3 (30.2) pg in females. The range of MCHC was 263 - 375 (335) g/L in males and 278 - 372 (325) g/L in females. The range of MCV was 83.9 - 99.1 (91.2) fl in males and 82.6 - 99.1 (91.3) fl in females. The range of HCT was 38.0% - 50.8% (44.8%) in males and 33.5% - 45.0% (38.9%) in females. The lower limits of WBC in Lanzhou and Chengdu were lower than those in other cities (P < 0.01). The concentrations of RBC and Hb in Kunming were significantly higher than those in other cities (P < 0.01). There was no significant difference in other parameters among different cities. The WBC, RBC, and Hb in the females were lower than those in the males (P < 0.01). CONCLUSIONS: The reference ranges of different parameters of venous blood of Chinese have been primarily established. There is little discrepancy in those reference range in different district (except for highland). Some of the indices such as WBC, MCH and MCV measured by analyzer are quite different from those measured manually.

[Change of T lymphocyte and its activated subsets in SARS patients].

OBJECTIVE: To study abnormal changes of T lymphocyte and its activated subsets in severe acute respiratory syndrome (SARS) patients. METHODS: Flow cytometer with multi-color flouroscence and hematology analyzer were used to detect the expression of T lymphocyte and its activated a subsets in 240 SARS patients including 50 cases of critical type and 190 cases of common type. RESULTS: Statistical analysis by means of SAS software showed that there was significant decrease in absolute counts (AC) of T lymphocyte and its subsets in SARS patients when compared with normal people, while percentages (PC) of CD3+CD25+ and CD3+ HLA-DR+ subsets were increased markedly. Compared with common type, there was significant decrease in absolute counts of critical type of T lymphocyte, CD4+, CD25+CD3+, CD28+CD4+, and CD95+CD4+subsets. The ACs of T lymphocytes including CD4 and CD8 subsets in different phases were as below: III > II > I. The ACs of subsets involved in activation such as CD3+ HLA-DR+/lym, CD3+CD25+/lym, CD28+CD4+/CD4, CD28+CD8+/CD8, and CD38+CD4+/CD4 all were highest in group III. In addition, the AC and PC of CD95+CD4+/CD4 and CD95+CD8/CD8 subset in group III were highest while group I was lowest. CONCLUSIONS: With depressing cellular immunity, the activation of T lymphocytes were suppressed obviously in SARS patients, especially for critical patients.

[The relationship among the counts of platelet, thrombopoietin and spleen index in patients with liver cirrhosis].

OBJECTIVE: To determine the reason of thrombocytopenia in patients with liver cirrhosis, we studied the relationship among platelet counts, serum thrombopoietin (TPO) level and spleen index. METHODS: Serum TPO, platelet counts and spleen index were measured in 71 cirrhotic patients. TPO was measured with ELISA method, spleen index were measured on ultrasonography by the same doctor. RESULTS: Platelet counts in patients with cirrhosis were lower than that of healthy group [(109.20+/-53.39) vs (169.63+/-26.60) x 10(12)/L, P<0.05]. Serum thrombopoietin level in patients with cirrhosis was similar to that of healthy group [(436.42+/-258.97) vs (412.63+/-132.80) pg/ml, P>0.05]. However, serum thrombopoietin level decreased as liver disease aggravated, [(526.13+/-317.44) pg/ml in Child-Pugh grade A, (445.22+/-214.90) pg/ml in grade B and (311.45+/-182.66) pg/ml in grade C, grade A vs. Grade C, P<0.05]. However, decline in platelet counts was accompanied with incline in spleen index coordinately. 35 of 71 cirrhotic patients had normal platelet counts whereas 36 of them had thrombocytopenia. Thrombopoietin levels were higher in non-thrombocytopenia group than in thrombocytopenia group [(529.43+/-282.64) vs. (351.27+/-228.25)pg/ml, P<0.01]; but spleen index of two groups showed no difference [(29.65+/-12.00) vs. (36.35+/-12.68) cm2, P>0.05]. Correlation was found between thrombopoietin level and platelet counts (r=0.252, P=0.025); no correlation was found between spleen index and platelet counts (r=-0.238, P=0.062). CONCLUSION: The decline serum TPO levels might play an important role for thrombocytopenia in patients with liver cirrhosis.

[The contrast study of Pre-S1 protein, HBV-DNA and HBeAg in diagnosing viral replication in patients with chronic hepatitis B].

OBJECTIVE: To determine the role of Pre-S1 protein in diagnosing viral replication in patients with chronic hepatitis B. METHODS: 104 consecutive patients with chronic hepatitis B were included in the study, liver biopsy were performed in all patients. Serial serum samples were studied with the quantitative determination of HBV-DNA by a quantitative PCR assay, determination of Pre-S1 protein by ELISA. RESULTS: The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg HBeAg anti-HBc (+) both were 96.5%. The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg anti-HBe anti-HBc (+) were 81.5%, 72.3%, respectively. The positive rates of HBV-DNA and Pre-S1 protein in patients with HBsAg anti-HBc (+) were 87.5%, 75.0%, respectively. It represented some patients with HBeAg (-) anti-HBe (+/-) still had viral replication. HBV-DNA>10(3) copy/ml as positive criteria for diagnosing viral replication, the positive rate of HBeAg, Pre-S1 were 31.5% (28/89), 80.9% (72/89) in patients with HBV-DNA>10(3) copy/ml, respectively. The concordance rates of HBeAg, Pre-S1 with HBV-DNA were 40.0% (42/104), 82.0% (85/104), respectively. CONCLUSION: It showed that Pre-S1 was more sensitive than HBeAg in diagnosing viral replication in patients with chronic hepatitis B.