OBJECTIVETo compare the sensitivity and practicability of modified Bethesda assay and Nijmegen assay in detecting factor VIII (FVIII) inhibitor.

METHODSModified Bethesda assay and Nijmegen assay were used to screen FVIII inhibitors in 237 patients with hemophilia A. The buffer plus universal coagulation reference plasma (UCRP) was used to establish a standard curve for FVIII: C assay in modified Bethesda method, instead of Nijmegen plasma plus FVIII deficiency plasma in Nijmegen method. The cutoff value for positive FVIII inhibitors is > or = 0.6 BU/ml.

RESULTSThe positive rate of FVIII inhibitors was 5.5% (n = 13) when using modified Bethesda assay and was 8.4% (n = 20) when using Nijmegen assay (P > 0.05).

CONCLUSIONModified standard Bethesda assay is a convenient and feasible method for detecting FVIII inhibitors.

Adolescent ; Adult ; Autoantibodies ; blood ; Blood Coagulation Tests ; methods ; Child ; Child, Preschool ; Factor VIII ; immunology ; Female ; Hemophilia A ; blood ; immunology ; Humans ; Male ; Middle Aged ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo observe therapeutic effect of acupuncture at acupoints selected according to rehabilitation medical theory on upper limb spasticity in the patient of poststroke.

METHODSSixty cases were randomly divided into an acupuncture group and an electro-stimulation group, 30 cases in each group. The acupuncture group were treated by acupuncture at the contralateral scalp motor region of the affected limb, Jiquan (HT 1), Chize (LU 5), Daling (PC 7) on the flexor side and Jianyu (LI 15), Tianjing (TE 10), Yangchi (TE 4) on the extensor muscle side of the affected limb; the electro-stimulation group were treated by electric stimulation. The two groups also were treated with necessary medical treatment and anti-spasm rehabilitation motor training. The course was 3 weeks. Modified Ashworth Scale for muscle spasm (MAS), modified Fugl-Meyer Assessment (FMA) for upper limb motor function, and Modified Barthel Index (MBI) for ability of daily living were used for assessment of the therapeutic effect.

RESULTSAfter treatment, the spasm was significantly alleviated, the motor function of the upper limb and daily living ability were significantly increased (P<0.01) in the two groups; after treatment, BMI scores in the acupuncture group was very significantly superior to that in the electro-stimulation group. The total effective rate was 93.3% in the acupuncture group and 86.7% in the electro-stimulation group, with no significant difference between the two groups.

CONCLUSIONProper acupuncture is an effective method for upper limb spasm in the patient of poststroke, and the therapeutic effect is better for mild-moderate spasm of the upper limb.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Muscle Spasticity ; therapy ; Stroke Rehabilitation

OBJECTIVETo investigate the mechanism of anticoagulation protein defect in the pathogenesis of unexplained recurrent miscarriage.

METHODSFifty-seven patients with a history of unexplained abortion were enrolled as the investigation group for tests of protein C, protein S, antithrombin III (AT-III), as well as activated protein C resistance (APC-R). The control group consisted of fifty healthy women with a history of normal pregnancy and delivery. Blood samples were obtained for, measuring serum activity of protein C, protein S, AT-III, and APC-R. Patients with positive APC-R were tested for factor V (FV) Leiden gene mutation by PCR-RFLP method.

RESULTSOf the 57 patients, 12 (21.1%), 1 (1.8%), and 5 (8.8%) cases were found with protein S, protein C, and AT-III deficiency respectively, and 13 (22.8%) cases with positive results of APC-R. Of the control group, no protein C or AT-III deficiency was ever found, whereas 2 (4.0%) volunteers were presented with protein S deficiency and 3 (6.0%) with positive results of APC-R. No FV Leiden gene mutation was identified in all the patients with positive APC-R results. Late spontaneous abortion cases had higher incidence of anticoagulation protein defect than the early cases.

CONCLUSIONAnticoagulation protein defect may play a role in the pathogenesis of fetal loss, especially for those occurring in late stage of pregnancy.

Abortion, Habitual ; blood ; etiology ; Activated Protein C Resistance ; blood ; complications ; genetics ; Adult ; Antithrombin III ; metabolism ; Antithrombin III Deficiency ; blood ; complications ; Factor V ; genetics ; Female ; Humans ; Point Mutation ; Protein C ; metabolism ; Protein C Deficiency ; blood ; complications ; Protein S ; metabolism ; Protein S Deficiency ; blood ; complications

Objective:To observe the possible toxicity of long-term intravenous injection of Tanreqing injection in Beagle dogs， so as to provide experimental data for its clinical safe medication. Method:A total of 32 Beagle dogs （16 males and 16 females） were randomly divided into the low- （2.5 mL·kg^-1）， medium- （5.0 mL·kg^-1）， and high-dose （10.0 mL·kg^-1） Tanreqing injection groups and control group according to their body mass indices， with eight dogs in each group. In the waking state， the dogs were treated with intravenous injection of corresponding drugs into the medial cephalic vein of forelimb for 13 weeks， followed by four-week drug withdrawal. After the observation of general condition， body mass， and food consumption， the Beagle dogs were subjected to electrocardiography， ophthalmoscopy， hematological examination， serum biochemistry， and blood coagulation test in the middle of medication （week 6）， at the end of medication （week 13）， and during recovery （week 17）. Then the gross anatomy was conducted for calculating the major organ coefficients and observing the histopathological changes. Result:No obvious toxic reaction was found in each group， but the decreased fibrinogen and increased Kupffer's cells phagocytizing yellow-brown pigment in hepatic sinusoids were observed in the high-dose Tanreqing injection group following three months of medication. Reduction of fibrinogen was not observed in recovery period， but Kupffer's cells that phagocytized yellow-brown pigment still existed. Conclusion:The intravenous injection of Tanreqing injection at 2.50 mL·kg^-1 （low dose）， 5.00 mL·kg^-1 （medium dose） or 10.00 mL·kg^-1 （high dose） for three months in Beagle dogs resulted in no obvious toxic reaction. However， it is still suggested to test the liver function and blood coagulation after long-term administration of high-dose Tanreqing injection.

OBJECTIVETo screen for factor VIII inhibitor in patients with hemophilia A (HA) and explore the environmental risk factors for inhibitor development.

METHODSTotally 265 patients with HA were enrolled, including 107 consecutive inpatients and outpatients in Peking Union Medical College Hospital from April 2003 to April 2007 and 158 patients newly recruited from other hospitals. FVIII: C activity was measured by one-stage coagulation assay. FVIII inhibitor was determined using Bethesda method.

RESULTSIn 265 HA patients, FVIII inhibitor was detected in 22 patients (8.3%). Nine of them (86.4%) were low responders (inhibitor titers < or = 5 000 BU/L), 3 (13.6%) were high responders (the titers > 5 000 BU/L). The frequency of FVIII inhibitor was 50% in the patients aged over 50 years, which was significantly higher than those in other age groups (P = 0. 000). Among 158 newly recruited patients with full clinical data, the frequency of FVIII inhibitor was 12.8% in patients who had received infusion of FVIII products for more than 12 doses on average each year, while it was 5.8% in whom the infusion doses were less than 12 (P = 0.156). The frequency of FVIII inhibitor was 28.5% in patients with a history of continuous infusion of FVIII products whereas it was only 1.6% in patients without such history (P = 0.000). In patients who exposed to multiple-branded or single-branded FVIII products, the frequencies of FVIII inhibitor were 9.3% and 3.9%, respectively (P = 0.229).

CONCLUSIONThe development of factor VIII inhibitor in patients with hemophilia A may be related to the age and the history of continuous infusion of FVIII products.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Environment ; Factor VIII ; antagonists & inhibitors ; Hemophilia A ; blood ; Humans ; Infant ; Male ; Middle Aged ; Risk Factors ; Young Adult

OBJECTIVETo investigate the relationship of angiogenesis and the Ang family members/ receptor (Ang/Tie2) in hemangioma.

METHODSExpression of Ang1, Ang2 and the receptor Tie2 was detected with immunohistochemical SP method and RT-PCR method in 17 cases of proliferating hemangioma, 13 involuting cases and 10 cases of normal children skin.

RESULTSThe expression of Ang2 and Tie2 was higher markedly in proliferating hemangiomas than in involuting hemangiomas (P < 0.01), and was rare or negative in normal skin. Expression of Ang1 was rare or negative both in hemangioma and normal skin without significant difference between them (P > 0.05).

CONCLUSIONAng/Tie2 system may play an important role in the proliferating and involuting process of hemangioma.

Angiopoietin-1 ; metabolism ; Angiopoietin-2 ; metabolism ; Child, Preschool ; Hemangioma ; metabolism ; pathology ; Humans ; Infant ; Receptor, TIE-2 ; metabolism

BACKGROUNDHemophilia A (HA) is an X-linked inherited bleeding disorder caused by decreased activity of factor VIII (FVIII) due to heterogenous mutations in the FVIII coding gene (F8). The type of mutation plays an important role in the FVIII inhibitor formation. To date, several studies on the spectra of F8 defects have been performed in Western populations, but similar studies in Asian races are scarce. Here, we reported the distribution of the F8 gene mutations in 18 unrelated Chinese patients with HA.

METHODSIntron 22 and intron 1 inversions in the F8 gene were screened in 158 unrelated patients with HA using a long-distance PCR and multiplex PCR method. Direct sequencing of the coding region of the F8 gene was used to identify the mutations responsible for HA in 18 unrelated Chinese HA patients who were negative for intron 22 and intron 1 inversions; sequences were compared with the HAMSTeRS database. A clotting method was used to assay the FVIII activity level and the Bethesda assay was used to detect the FVIII inhibitor.

RESULTSA total of 18 different HA F8 mutations were identified, seven of which were described for the first time. These novel mutations included five small deletions, one point mutation and one small insertion. One novel mutation (4382-3 AC deletion) was associated with inhibitor development.

CONCLUSIONThese data extend our insight into the mechanisms by which novel amino acid mutations may lead to HA and how the HA patient genotypes influence the risk of FVIII inhibitor.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; Factor VIII ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Hemophilia A ; genetics ; Humans ; Infant ; Introns ; genetics ; Male ; Middle Aged ; Mutation ; Point Mutation ; genetics ; Polymerase Chain Reaction ; Young Adult

OBJECTIVESTo explore the frequency, clinical features and risk factors of venous thromboembolism (VTE) in hospitalized patients.

METHODSThe frequency, demographic features, and acquired and inherited factors of in-patient cases of VTE in Peking union medical college hospital from 1994 to 2004 were analyzed retrospectively.

RESULTSSix hundred and seventy-two patients were enrolled. Among them, male to female ratio was 1.2 and the median age was 53 (14 - 92). Five hundred and eighty (86.3%) patients were at their first diagnosis with the peak ages between 40 and 50 for men and 50 and 60 for women. More common acquired risk factors were antiphospholipid antibody syndrome (APS) (32.0%), trauma / surgery (31.1%) and malignancies (17.1%). 35.7% of the patients had multiple acquired risk factors. Before the initiation of anticoagulation therapy, the activities of protein C (PC), protein S (PS) and antithrombin (AT) were measured in 94 patients. The deficiency of these three natural anticoagulants was 44.7%. Among the anticoagulant deficiencies, PC deficiency was the commonest one (13.8%). Combined deficiency of PC and AT accounted for 10.6%. 31.6% of the 94 patients had inherited plus acquired risk factors.

CONCLUSIONSAge for the first event of VTE in the men was about 10 years ahead of that in the women. The major acquired risk factors were APS, trauma/surgery and malignancies, and inherited risk factors were PC deficiency and PC + AT combined deficiencies. It seems that the coexistence of multiple risk factors plays an important role in triggering VTE.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Venous Thromboembolism ; etiology

Objectives: To assess the diagnostic values of latex immunoturbidimetric assay (LIA) and particle immunofiltration assay (PIFA) for heparin-induced thrombocytopenia (HIT) . Methods: Samples from 94 patients with suspected HIT from May 2016 to July 2018 in our hospital were prospectively analyzed by the two immunoassays. Their medical records and further follow-up data were also collected and analyzed by our hematologists to make the 4Ts scores and confirm the diagnosis of HIT, respectively. Performance characteristics of the two immunoassays were assessed, including sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) . Their post-test probabilities (PTP) were also calculated based on the 4Ts score. Results: Among 94 cases, 15 (16.0%) had a positive HIT, including 6 of 37 (16.2%) with an intermediate, and 9 of 15 (60.0%) with a high 4Ts score. PIFA operating characteristics were: sensitivity 100.0% (15/15) , specificity 51.9% (41/80) , PPV 28.3% (15/53) , NPV 100.0% (41/41) . The positive PTP in intermediate and high 4Ts score group were 28.7% and 75.7%, respectively, while negative PTP were all 0. At manufacturers' cutoffs, LIA operating characteristics were: sensitivity 66.7% (10/15) , specificity 94.9% (75/79) , PPV 71.4% (10/14) and NPV 93.8% (75/80) . The positive and negative PTP in intermediate 4Ts score group were 71.8% and 6.3%, while 95.2% and 34.4% in high 4Ts score group, respectively. Receiver operating characteristic (ROC) analysis manifested that LIA was preferable than PIFA, and combining the 2 assays together was significantly better than single test. Conclusions: 4Ts score is still an important tool for the diagnosis of HIT. Combining clinical score with heparin/PF4 antibody assay can increase the accuracy of confirming or excluding HIT. Although PIFA is inferior to LIA in the diagnostic value, its user friendliness and 100% NPV have major advantages. Combining the 2 assays together can achieve a higher diagnostic value.
Antibodies ; Anticoagulants ; Enzyme-Linked Immunosorbent Assay ; Heparin/adverse effects* ; Humans ; Immunoassay ; Platelet Factor 4 ; Thrombocytopenia/chemically induced*