Evaluation of a single-tube multiplex polymerase chain reaction screen for detection of common alpha-thalassemia genotypes in a clinical laboratory

We prospectively compared a single-tube multiplex polymerase chain reaction (PCR) for detecting alpha-thalassemia with our current approach using 452 blood samples. Initial evaluation of 89 specimens revealed sensitivity and specificity, respectively, for the hemoglobin H inclusion body test (HbH prep) vs PCR for detecting alpha0-thalassemia carriers of 0.79 and 0.96 and for a mean corpuscular volume (MCV) of 82 microm3 (82 fL) or less, 1.0 and 0.45. Detection of all alpha-thalassemia genotypes was significantly lower for HbH prep and MCV (sensitivity and specificity, respectively: HbH prep, 0.48 and 0.96; MCV, 0.87 and 0.47). In a follow-up evaluation of patients with positive HbH prep results or suspected alpha-thalassemia prescreened by low MCV, the sensitivity and specificity, respectively, of HbH prep vs PCR increased to 0.97 and 0.93 for alpha0-thalassemia and 0.83 and 0.92 for any alpha-thalassemia. PCR detected alpha-thalassemia in 37.2% of 298 suspected alpha-thalassemia cases with suggestive indices but negative HbH prep results and no detectable hemoglobinopathy. This multiplex approach was more sensitive than the HbH prep for detecting all alpha-thalassemia genotypes, particularly alpha+-thalassemia; was particularly valuable for identifying carriers of alpha0-thalassemia at risk for offspring with hemoglobin Bart hydropsfetalis, regardless of other diagnosed hemoglobinopathies; and is an ideal adjunct to standard clinical screening protocols for detecting alpha-globin deletions.     

Isolation of human complement subcomponents C1r and C1s in their unactivated, proenzyme forms

We have modified a standard isolation procedure for C1r and C1s, which employs IgG-Sepharose affinity chromatography followed by DEAE chromatography. As usual, all steps were performed at low temperature and two proteolytic inhibitors, PMSF and NPGB, were added during affinity chromatography on IgG-Sepharose. The novel condition was to keep the pH at pH 6.1 during the entire procedure, where activation was markedly depressed. In addition, purification was improved by washing the IgG-Sepharose column with a buffer free of added divalent cations immediately prior to elution of the C1r and C1s with EDTA. The final yields of highly purified C1r and C1s were about 20%; little or no activated material was detected in these highly purified fractions.     

中医灸与人体穴位红外辐射光谱特性研究

本文通过锁相放大技术测到人体不到1cm^2面积的体表红外辐射光谱。通过对这些光谱的归一化处理后发现，不同人体和同一人体的不同穴位，尽管其红外辐射绝对光谱及强度相差很大，但归一化光谱都有相当的一致性与可比性。进一步将人体发射的平均归一化光谱与中医艾条灸、隔姜灸、隔蒜灸及隔附子饼灸点燃过程中的光谱比较发现，后面三种间隔灸与人体自发辐射的光谱有着惊人的一致性，而艾条灸等的光谱却相差甚远。     

Molecular subtyping of Vibrio cholerae O1 and O139 by pulsed-field gel electrophoresis in Hong Kong: correlation with epidemiological events from 1994 to 2002

Two hundred twenty isolates of Vibrio cholerae O1 and O139 collected from 1994 to 2002 in Hong Kong were analyzed by pulsed-field gel electrophoresis (PFGE). Chromosomal DNAs from all V. cholerae isolates in agarose plugs were digested with the restriction enzyme NotI, resulting in 20 to 27 bands. Sixty distinctive PFGE patterns in the range of 10 to 300 kb were noted among 213 isolates typeable by PFGE. By comparing the common PFGE patterns obtained from four well-defined outbreaks of V. cholerae O1 and O139 with those obtained from other, epidemiologically unrelated isolates during the study period, indistinguishable and similar PFGE patterns were identified, indicating their close relatedness, in agreement with the results of epidemiological investigations. Heterogeneous PFGE patterns (with four to six banding differences), however, were identified among strains that were imported from other parts of Asia, including Indonesia, India, and Pakistan. Correlations with epidemiological information further support the usefulness of PFGE as an epidemiological tool in laboratory investigations of suspected outbreaks. Standardization of PFGE methodology will allow international comparison of fingerprint patterns and will form the basis of a laboratory network for tracking V. cholerae.     

Comparison of hospital staff performance when using desk top analysers for "near patient" testing.

The quality and reliability of four desk top analysers were evaluated. In the context of an outpatient clinic, intensive care unit, and a mock up of a physician's office. Seventeen nurses, 14 physicians, and 12 medical office personnel took part in the study. The instruments and tests evaluated were Reflotron (glucose, cholesterol, triglycerides, gamma-glutamyl transferase), Seralyzer (creatinine, glucose, potassium, aspartate aminotransferase), Vision (glucose, (creatinine, glucose, potassium, aspartate aminotransferase), Vision (glucose, urea, cholesterol, triglycerides alkaline phosphatase, uric acid), and DT60 (sodium, potassium, glucose, amylase, uric acid and creatinine). Of the 320 tests performed on the Vision, only two differed by more than 10% between the specialist staff and other groups. For those performed on the Seralyzer, 95 of 254 results differed by more than 10%, 19 of 199 by more than 10% for the Reflotron, and 50 of 318 by more than 10% for the DT60. In general, the nurses were more adept at using the analysers than the physicians and medical office personnel.     

Cytocidal and apoptotic effects of the ClyA protein from Escherichia coli on primary and cultured monocytes and macrophages

Cytolysin A (ClyA) is a newly discovered cytolytic protein of Escherichia coli K-12 that mediates a hemolytic phenotype. We show here that highly purified ClyA and ClyA-expressing E. coli were cytotoxic and apoptogenic to fresh as well as cultured human and murine monocytes/macrophages.