[Investigation and analysis of health-related quality of life in myasthenia gravis patients with myasthenia gravis quality of life-15 Chinese version].

OBJECTIVE: To investigate the level and influencing factors of health-related quality of life in myasthenia gravis (MG) patients with myasthenia gravis quality of life-15 (MGQOL-15) Chinese version and to provide corresponding measures in one tertiary hospital of Sichuan Province. METHODS: We collected the general data (gender, age, body mass index BMI, marital status, educational level and employee status), clinical data [Osserman type, myasthenia gravis composite (MGC), other immunopathies, disease duration, frequency of outpatient visits per month, ratio of disease cost to income each month and frequency of symptoms during the past month] and the MGQOL-15 Chinese version from 168 myasthenia gravis patients in one tertiary hospital of Sichuan Province. RESULTS: The mean score of MGQOL-15 was 17.67±12.78. The score of the item "My occupational skills and job status have been negatively affected." was the highest, followed by "I have trouble using my eyes." and "I am frustrated by my MG." Single factor analysis showed that MG patients' QOL were different with different disease severity MGC (F=19.353, P<0.001), ratio of disease cost to income each month (F=5.831, P<0.001) and the frequency of symptoms during the past month (F=9.128,P<0.001). Multiple regression analysis showed that disease severity MGC (ß=0.743,P<0.001), ration of disease cost to income each month (ß=3.347,P<0.001) and the frequency of symptoms during the past month (ß=2.216,P<0.003) were the main predictors of HRQOL in the MG patients. CONCLUSION: Our study showed that the MGQOL-15 is helpful for clinicians to evaluate MG patients' QOL regularly, investigate the influencing factors and implement corresponding interventions the so as to improve the patients' quality of life. Disease severity MGC, ratio of disease cost to income each month and the frequency of symptoms during the past month were the main predictors of MG patients' QOL. Clinicians should pay more attention to MG patients' disease severity MGC and the frequency of symptoms during the past month.

Detection of fetal sex in the peripheral blood of pregnant women.

OBJECTIVE: To choose the best method to examine fetal sex in maternal blood as early as possible and evaluate the quantitative change of fetal-free DNA in maternal plasma. METHOD: One hundred and fifty pregnant women were studied at 5-9 completed weeks of gestation. Fetal cells were isolated using lymphocyte separation liquid and 3% gelatin. Furthermore, fluorescence in situ hybridization was used to examine the terminal of the Y chromosome. Nested polymerase chain reaction (PCR) and real-time fluorescence quantitative PCR (FQ-PCR) were used to amplify the SRY gene of the plasma DNA extracted from the same 150 samples of maternal blood. Sequential analysis was performed using FQ-PCR during the whole pregnancy on 32 pregnant women carrying male fetuses. RESULTS: Using fluorescence in situ hybridization, we can find male fetal cells in maternal blood as early as the 49th day. We can also find free fetal DNA in maternal plasma as early as the 49th and the 42nd day of pregnancy using nested PCR and FQ-PCR. The amount of fetal DNA was increasing with the gestation week. The standard value of every gestation week was obtained by FQ-PCR. CONCLUSION: FQ-PCR was the best method to detect fetal sex in early pregnancy. There is a principle of quantitative change of free fetal DNA in maternal plasma.

Methylation status of the fragile histidine triad and E-cadherin genes in plasma of cervical cancer patients.

Recent evidence suggests that tumor cells may release DNA into the serum and plasma of afflicted cancer patients. However, no report existed regarding the methylation status of the fragile histidine triad (FHIT) and E-cadherin genes in plasma samples of cervical cancer patients. Methylation-specific PCR (MSP) was employed to examine CpG island methylation of the FHIT and E-cadherin genes in 151 pretreatment plasma samples and 30 tumor tissue samples from cervical cancer patients. MSP products were cloned and sequenced. CpG island methylation of the FHIT and E-cadherin genes was detected in 30.46% and 39.74% of plasma samples, respectively, and in 53.33% and 60.0% of tissue samples, respectively. The total concordance rate of methylation between plasma samples and tissue samples in FHIT gene was 80.00% and that in E-cadherin gene was 76.66%. At least one of the two methylated genes was detected in 56.29% of plasma samples and 76.7% of tissue samples. The presence of both methylated genes was detected in 13.9% of plasma samples and 36.67% of tissue samples. We found that the higher the clinical stage and histologic grade, the higher the rate of methylation in both genes in plasma samples. CpG island methylation of the FHIT and E-cadherin genes is present in plasma of cervical cancer patients. Using the two genes as markers simultaneously may allow clinicians to diagnose and evaluate the effect of treatment earlier and using fewer invasive procedures.

Construction and identification of a single stranded cDNA clone containing full-length genome of hepatitis G virus.

AIM: To construct a single cDNA clone with full-length genome of hepatitis G virus (HGV) could be transcribed and expressed in vitro. METHODS: The 5 initial HGV cDNA fragments of Iw5, Iwq2, Iwh6, Iw3 and Iw3 used in this study were amplified from serum of a Japanese non A-E hepatitis patient. These fragments overlapped and covered the entire genome from 5'-end to 3'-end of HGV cDNA. Overlap extension PCR and ligation methods were used with 12 primers for the construction of a full-length genomic HGV cDNA clone from the subgenomic fragments. RESULTS: A single HGV cDNA clone (pHGVqz) was successfully constructed, physical mapping of the generated pHGVqz found identical to what we expected, and the sequence was deposited with the GenBank under the Accession number AF081782. The analysis of the full-length sequence, which was able to be in vitro transcribed and expressed, showed that this single clone contained 9373 nucleotides (encoding 2873 amino acids), and shared high homologies with other compared HGV isolates. CONCLUSION: A full-length genomic HGV cDNA clone is generated for the first of the kind in this study, it could be expressed and transcripted. This single cDNA clone is expected to be of importance in the investigation on replication and pathogenicity of HGV.