OBJECTIVETo establish the linkage methods of X ba I polymorphisms specific for FVIII gene intron 22, and to find a rapid and simple system for haemophilia A (HA) carrier detection and prenatal diagnosis.

METHODSA long PCR to amplify FVIII gene intron 22 followed by X ba I digestion was used to assay the gene rate and heterozygosity rate of 206 unrelated people. Detection of intron 22 inversion by long distance PCR (LD-PCR) and XbaI, BclI, Hind III, DXS52, STR polymorphism within intron 13 and 22 by hereditary linkage analysis were assays in 20 HA pedigrees.

RESULTSThe gene rate and polymorphism information contents of 206 people were 0.5475 and 0.4955 respectively, 7 of 20 HA families were diagnosed as intron 22 inversion, 6 of 13 non-inversion HA families were diagnosed by X ba I linkage analysis, 8 of 13 non-inversion HA families were diagnosed by two or more linkage analysis.

CONCLUSIONSThe improved X ba I linkage analysis is a specific and useful molecular diagnosis marker. LD-PCR and five-linkage analysis can be used in prenatal HA gene diagnosis.

Factor VIII ; genetics ; Female ; Genetic Linkage ; Genotype ; Hemophilia A ; diagnosis ; genetics ; Heterozygote ; Humans ; Introns ; Polymorphism, Single Nucleotide ; Pregnancy ; Prenatal Diagnosis ; methods

OBJECTIVETo elucidate the therapeutic effect and the influence on PI3K-Akt-PKB-BAD-CREB-PCREB pathway in focal cerebral ischemia rat responses before and after treatment with baicalin and jasminoidin given alone or in combination.

METHODRat model of ischemia reperfusion was established with thread. Generally accepted methods were used, including TTC staining, behavior test, as well as micro and ultra microscopy which can dynamically and accurately monitor pathological and physiological changes after cerebral ischemia on earlier period, to evaluate the brain injury induced by ischemia and the attenuations by the drugs. The difference of PI3K-Akt-PKB-BAD-CREB-PCREB expression was detected by western-blot technology.

RESULT AND CONCLUSIONThe combination of baicalin and jasminoidin composition can be potential neuroprotective agent. TTC staining technology combined with behavior grade and ultrmicro-structure observation on brain tissue is effective method to evaluate protective agent, which is related to signal transduction PI3K-Akt-PKB-BAD-CREB-PCREB pathway. The results provide benofical basis for revealing the complex of therapeutic mechanism of traditional Chinese medicine Qingkai Ling (QKL).

Animals ; Behavior, Animal ; drug effects ; Brain ; pathology ; Brain Ischemia ; complications ; metabolism ; pathology ; CREB-Binding Protein ; metabolism ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Drug Combinations ; Flavonoids ; isolation & purification ; pharmacology ; Gardenia ; chemistry ; Injections ; Iridoids ; isolation & purification ; pharmacology ; Male ; Neuroprotective Agents ; pharmacology ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-akt ; metabolism ; Pyrans ; isolation & purification ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; pathology ; Scutellaria ; chemistry ; Signal Transduction

OBJECTIVETo improve the accuracy of preoperative evaluation of cervical lymph node metastasis in thoracic esophageal squamous carcinoma.

METHODSForty-two patients with thoracic esophageal squamous carcinoma underwent neck ultrasonography. Enlarged lymph nodes with their long axis greater than 10 mm and a short-to-long axis ratio greater than 0.5 were considered as metastatic.

RESULTSPreoperative neck ultrasonography revealed the enlarged lymph nodes in 16 patients, but only in 5 (31%) cases the nodes were palpable. Among them 9 were classified as metastatic (cM(1-LN)), including 4 patients with palpable nodes. In 5 cM(1-LN) patients surgical intervention was canceled and the remaining 37 patients underwent trans-thoracic esophagectomy. Cervical node metastasis (pM(1-LN)) was confirmed pathologically in 6 surgical patients, 4 with tumors invading the adventitia (pT3) and the other 2 into the surrounding structure (pT(4)) (pT(1), pT(2) vs. pT(3), pT(4), P = 0.020). All 6 pM(1-LN) patients had concomitant mediastinal node metastasis and 4 of them had upper abdominal node metastasis. Statistically significant relationship was detected between cervical and abdominal nodal status (r = 0.536, P = 0.007). In comparison with the results of pathological examination and treatment response, the accuracy and sensitivity were 81% and 95% (P = 0.043), 36% and 82% (P = 0.081), respectively, for palpation and ultrasonography. Five out of 39 (13%) patients had their therapy changed due to ultrasonographic findings.

CONCLUSIONSNeck ultrasonography for cervical lymphadenopathy is of high sensitivity and accuracy, which plays an important role in the preoperative evaluation and therapeutic decision-making.

Adult ; Aged ; Carcinoma, Squamous Cell ; diagnostic imaging ; secondary ; surgery ; Esophageal Neoplasms ; pathology ; surgery ; Female ; Head and Neck Neoplasms ; diagnostic imaging ; secondary ; surgery ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; diagnostic imaging ; Lymphatic Metastasis ; diagnosis ; Male ; Middle Aged ; Neck ; diagnostic imaging ; Sensitivity and Specificity ; Ultrasonography

The aim of this study was to explore application value of detecting platelet associated antibody and platelet membrane glycoprotein in the diagnosis and prognosis for immune thrombocytopenia. The platelet associated immunoglobulin (PAIg) and platelet membrane glycoprotein (CD41, CD61, GPIIb/IIIa) in 76 cases of immune thrombocytopenia and 30 healthy subjects were determined by FCM. The results showed that PAIg level in ITP patients included PAIgG (31.25 +/- 18.06)%, PAIgM (32.41 +/- 15.51)%, PAIgA (23.39 +/- 16.67)% which were remarkedly higher than in health control (10.48 +/- 5.05)%, (9.40 +/- 4.42)% and (7.23 +/- 3.61)% (P < 0.001). In patients with secondary immune thrombocytopenia (chronic aplastic anemia, SLE, Evans syndrome, liver cirrhosis hypersplenism, etc), PAIg level was higher than that in control group, while the platelet membrane glycoprotein in the blood of these patients was lower than that in control group. The level of PAIg decreased (P < 0.05) after treatment, but platelet membrane glycoprotein increased (P < 0.01). The result suggested that measurements for platelet membrane glycoprotein and platelet associated antibody by FCM were practical with high sensitivity, rapidity and simplicity used as a routine method in diagnosis and evaluation of the therapeutic effects in immune thrombocytopenia patients.
Adolescent ; Adult ; Aged ; Blood Platelets ; immunology ; Child ; Female ; Flow Cytometry ; Humans ; Immunoglobulins ; blood ; Integrin beta3 ; analysis ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; analysis ; Platelet Membrane Glycoprotein IIb ; analysis ; Platelet Membrane Glycoproteins ; analysis ; Purpura, Thrombocytopenic, Idiopathic ; blood ; diagnosis ; Thrombocytopenia ; blood ; diagnosis

Objective To analyze the impact of attack frequency as well as therapy strategies on outcome of patients with vasovagal syncope (VVS).Methods A total of 159 patients (aged from 15-59 years old) with VVS were included in this study.Patients were divided into low frequency (＜ 3) group (n =95) and high (≥3) frequency group (n =64) according to the attack frequency in the past 5 years at the primary survey.Patients received one of the three therapies:no treatment,physical therapy,and comprehensive treatment.All cases were followed up with telephone or outpatient visit for 24 months.Results Incidence of syncope was significantly higher in the high frequency group and in the low frequency group [40.6％ (26/64) vs.11.6％ (11/95),P ＜0.01].The overall improvement rate was significantly higher in the low frequency group than that of high frequency group (P ＜ 0.01).Improvement rate was significantly higher in the physical therapy subgroup and the comprehensive treatment subgroup than no treatment subgroup for patients with low attack frequency [81.8％ (27/33) vs.47.1％ (8/17),P ＜ 0.05 ; 82.2％ (37/45) vs.47.1％ (8/17),P ＜ 0.05],and in comprehensive treatment subgroup than in physical therapy subgroups observed between and [62.2％ (28/45) vs.31.6％ (6/19),P ＜ 0.05] for patients with high attack frequency.Conclusion Outcome is related to previous attack frequency for patients with VVS,physical therapy is effective for reducing the recurrence rate of syncope in VVS patients with low attack frequency while physical therapy combined with pharmacotherapy should be applied for VVS patients with high attack frequency to improve outcome.

OBJECTIVETo investigate the significance of mutation and single nucleotide polymorphism (SNP) of class III receptor tyrosine kinases such as PDGFRbeta and SHIP in acute myeloid leukemia (AML) patients.

METHODSScreening of the mutation and SNP of PDGFRbeta and SHIP by genomic PCR, RT-PCR, directly sequencing and Mass-ARRAY system was carried out in 273 AML patients.

RESULTSThe mutations of PDGFRbeta R685C and SHIP Q1153L were detected for the first time in AML patients. The positivity ratio was 0.73% and 0.36% respectively.

CONCLUSIONThe mutations of PDGFRbeta R685C and SHIP Q1153L may contribute to leukemogenesis of AML.

Humans ; Inositol Phosphates ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Mass Spectrometry ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Receptor, Platelet-Derived Growth Factor beta ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To design and manufacture a multifunctional cushion for venous thrombosis prevention of lower extremities, which can be also applied to the low extremities needing raising. Methods The cushion was made of anti-rust, low-weight, undeformable and easy-to-clean stainless material, which was composed of a pad and a massage apparatus. The pad had its height regulable from 16 to 30 cm and telescopic inclination from 30 to 70° to adapt the cushion to sizes of patients. There were 6 independent air chambers and tubular circulating pipes around the blanket wrapping the chambers so that extremity massage and temperature-controlled heating could be executed based on setting up pressure, time and temperature of the electromagnetic air and water pumps. Results The cushion simulated the functions of the muscle pump, which formed step-by-step-increasing pressure changes by driving the chambers to be inflated and discharged continuously to execute extremity massage, temperature-controlled heating etc. Conclusion The cushion gains advantages in flexible composition, patient comfort etc, solves the problems in fixing angle and height, constant time for pump inflation and discharge as well as temperature-controlled heating, and thus is worthy promoting clinically.

OBJECTIVETo establish a simple and rapid system for carrier detection and prenatal diagnosis in hemophilia A (CHA) family.

METHODSLong distance-polymerase chain reaction (LD-PCR) was selected for detection factor VIII intron 22 inversion. Polymorphism of factor VIII intragenic restriction fragment length polymorphism (RFLP) of Xba I and Hin d III, short tandem repeat (STR) within intron 13 and 22, as well as extragenic DXS52 (ST 14) variable number of tandem repeat (VNTR) were assayed by PCR and linkage analysis.

RESULTSSeventy-one females were diagnosed as carriers within 52 HA families. Twenty-one families were diagnosed to be factor VIII intron 22 inversion and 28 families were diagnosed by linkage analysis, whereas 3 families could not been diagnosed. Seventeen of 18 fetuses at risk were male. Ten of 17 male fetuses were shown to be affected and were subsequently aborted. Seven male fetuses were diagnosed to be not affected. One female fetus was identified to be HA carrier. One-year follow-up study demonstrated that these babies were normal and living well.

CONCLUSIONLD-PCR combined with multiple locus linkage analysis enables the direct and indirect detection of HA for carrier testing and prenatal diagnosis.

Factor VIII ; genetics ; Family Health ; Female ; Hemophilia A ; diagnosis ; genetics ; Humans ; Male ; Minisatellite Repeats ; genetics ; Pedigree ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Pregnancy ; Prenatal Diagnosis ; methods

OBJECTIVETo establish an automatic, high throughput, quick detection method of alpha thalassemia.

METHODSThe genotypes of -alpha(4.2) and -alpha(3.7) were detected by two real-time fluorescence PCRs using SYBR-Green1 (SYBR-PCR) with the analysis of dissociation curve (DC) and melting temperature (Tm). The PCR products were recombined into T-vector and the correct cloning was selected as positive control. Sensitivities were gained from a serious dilution of the recombinant as SYBR-PCR template, from which detection deadlines for two kinds of alleles could be determined. Totally 110 samples were detected by this technique.

RESULTSThe length of product of -alpha(4.2) and -alpha(3.7) were 1.65 kb and 1.9 kb respectively, and the Tm were (81.5+/-0.5) degrees C and (82.5+/-0.5) degrees C respectively. The detection deadline were 9 x 10(2 ) copies and 4.3 x 10(2) copies respectively. The sensitivity of the technique was much higher than that of regular PCR plus gel electrophoresis method, and the detection results of the technique were the same as that of multiplex PCR.

CONCLUSIONThe genotypes of -alpha(4.2), -alpha(3.7), non-deletion alpha alpha/and --(SEA) can be sensitively, exactly diagnosed by the real-time PCR with SYBR-Green1 combined with the dissociation curve analysis. The assay is automatic, low-cost, high throughput, and easy for quality control without fluorescence probe.

Genotype ; Humans ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity ; alpha-Thalassemia ; diagnosis ; genetics

To construct a safer and more efficient gene engineering Lactococcus Lactis for expressing phenylalaine ammonia lyase (PAL) which will be benefit for PKU therapy, pal cDNA of Parsly and synthesized sequence based on Lactococcus Lactis bias codons were recombined into two Lactococcus Lactis NICE systems. The activities of the expressed PAL were detected, and the effect of Lactococcus Lactis bias codons on the expression of exterior protein was analyzed. The results showed that the expression level of PAL was increased by using Lactococcus Lactis bias codons in both Lactococcus Lactis NICE systems. Through which several safer andmore efficient strains of the gene engineering Lactococcus Lactis were obtained.
Cloning, Molecular ; Codon ; genetics ; Genetic Vectors ; genetics ; Lactococcus lactis ; genetics ; metabolism ; Phenylalanine Ammonia-Lyase ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; metabolism ; Transformation, Bacterial