OBJECTIVEA molecular technique based on quasispecies analysis for tracing postexposure HIV transmission was applied in an investigation of a possible case of HIV transmission after blood transfusion.

METHODSSixteen plasma specimens were collected from 3 HIV infections (T1-T3) involved in a possible HIV transmission chain and 13 HIV/AIDS (C1-C13) controls. The RNAs were extracted and then amplified by RT-PCR, the PCR products were cloned and sequenced.BioEdit 6.0.7 and MEGA 4.0 software were used to analyze gene sequences, calculate gene dispersion ratio and construct phylogenetic tree.

RESULTSThe sequences of 13 specimens were successfully obtained.The HIV strains from T1, T2 and T3 were CRF07_BC recombinants, those from 5 out of the 6 controls lived in the same city with T2 and T3 were CRF07_BC recombinants as well, while those from 4 controls living in the same city with T1 were CRF01_AE recombinants. Compared with the clone sequences from T1, the mean gene dispersion ratio of T2 was the least (2.0%), followed by C12 (2.8%) , T3 (2.9%) and others. The phylogenetic tree showed that all clones from T1, T2, T3 and C12 might cluster together,and implied that the direction of HIV transmission was from T3 to T2, and then to T1.

CONCLUSIONThe results support the possible epidemiological clue that HIV was transmitted from T3 to T2, and then to T1, indicating that molecular epidemiological investigation could provide more direct evidence for tracing postexposure HIV transmission.

Female ; HIV ; genetics ; HIV Infections ; epidemiology ; genetics ; transmission ; Humans ; Male ; Molecular Epidemiology ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, RNA ; Transfusion Reaction

OBJECTIVETo use dried blood spot (DBS) in studying the sequence and subtype analysis of HIV-1 genome.

METHODS2 ml whole blood containing EDTA anticoagulant from 20 HIV-1 infected patients were collected, then 80 microl blood was used to propare DBS. QIAamp Blood Mini kit and 10% Chelex100 resin extracted DNA genome from whole blood and DBS as well as nested PCR amplified specifically HIV-1 Gp41 region from the two kinds of DNA extraction. Software MAGE 3.0 was used to study the sequence and subtype of the PCR products.

RESULTSEligible 18 paired samples were analysed to show that 16 of them belonged to C54A, 97CNGX-7F, 98CN006 subtypes. The other two samples might belong to B. CN._. RL42 and B. US. 90WEAU160.

CONCLUSIONData showed that there were parallel results between the whole blood and DBS samples including subtype analysis, position of mutation and types of amino acid sequencing. Since DBS itself facilitated the collection, transportation and storage, it could be used as a measure to collect blood sample in resource limited area and to develop molecular epidemiologic research as well as early diagnosis on infant exposed to HIV.

DNA, Viral ; blood ; HIV Infections ; blood ; genetics ; HIV-1 ; genetics ; Humans ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; methods

OBJECTIVETo compare the results of detecting HIV-1 load by using NucliSens HIV-1 QT and Amplicor HIV-1 monitor 1.5 assays.

METHODSEighty-two clinical samples were collected and HIV viral load was determined with the above-mentioned two methods.

RESULTSThe number of samples in which values obtained by NucliSens HIV-1 QT and Amplicor HIV-1 monitor 1.5 differed by <0.5 log10 RNA copies/ml and in which the viral load was undetectable accounted for 88.9 percent of the measures. The correlation coefficient between the two methods was 0.956 in 56 samples of Deltalog10 VL<0.5.

CONCLUSIONThe results of HIV-1 viral load determination with the two methods are highly comparable.

HIV Infections ; virology ; HIV-1 ; genetics ; isolation & purification ; Humans ; Nucleic Acid Amplification Techniques ; instrumentation ; methods ; RNA, Viral ; genetics ; Viral Load

BACKGROUNDRegulatory T-cells (Treg) play key roles in suppressing cell-mediated immunity in cancer patients. Little is known about perioperative Treg fluctuations in nonsmall cell lung cancer (NSCLC). Video-assisted thoracoscopic (VATS) lobectomy, as a minimal invasive procedure for treating NSCLC, may have relatively less impact on the patient's immune system. This study aimed to observe perioperative dynamics of circulating Treg and natural killer (NK) cell levels in NSCLC patients who underwent major lobectomy by VATS or thoracotomy.

METHODSTotally, 98 consecutive patients with stage I NSCLC were recruited and assigned into VATS or thoracotomy groups. Peripheral blood samples were taken on 1-day prior to operation, postoperative days (PODs) 1, 3, 7, 30, and 90. Circulating Treg and NK cell counts were assayed by flow cytometry, defined as CD4 + CD25 + CD127 low cells in CD4 + lymphocytes and CD56 + 16 + CD3- cells within CD45 + leukocytes respectively. With SPSS software version 21.0 (SPSS Inc., USA), differences between VATS and thoracotomy groups were determined by one-way analysis of variance (ANOVA), and differences between preoperative baseline and PODs in each group were evaluated by one-way ANOVA Dunnett t-test.

RESULTSIn both groups, postoperative Treg percentages were lower than preoperative status. No statistical difference was found between VATS and thoracotomy groups on PODs 1, 3, 7, and 30. On POD 90, Treg percentage in VATS group was significantly lower than in thoracotomy group (5.26 ± 2.75 vs. 6.99 ± 3.60, P = 0.012). However, a higher level of NK was found on all PODs except on POD 90 in VATS group, comparing to thoracotomy group.

CONCLUSIONSLower Treg level on POD 90 and higher NK levels on PODs 1, 3, 7, 30 in VATS group might imply better preserved cell-mediated immune function in NSCLC patients, than those in thoracotomy group.

Aged ; Carcinoma, Non-Small-Cell Lung ; immunology ; surgery ; Female ; Flow Cytometry ; Humans ; Killer Cells, Natural ; immunology ; Male ; Middle Aged ; Postoperative Period ; T-Lymphocytes, Regulatory ; immunology ; Thoracic Surgery, Video-Assisted ; methods ; Thoracotomy ; methods

OBJECTIVE: We wanted to report our experience of metallic stent placement after insufficient balloon dilation in graft hemodialysis patients. MATERIALS AND METHODS: Twenty-three patients (13 loop grafts in the forearm and 10 straight grafts in the upper arm) underwent metallic stent placement due to insufficient flow after urokinase thrombolysis and balloon dilation. The indications for metallic stent deployment included 1) recoil and/or kinked venous stenosis in 21 patients (venous anastomosis: 17 patients, peripheral outflow vein: four patients); and 2) major vascular rupture in two patients. Metallic stents 8-10mm in diameter and 40-80 mm in length were used. Of them, eight stents were deployed across the elbow crease. Access patency was determined by clinical follow-up and the overall rates were calculated by Kaplan-Meier survival analysis. RESULTS: No procedure-related complications (stent fracture or central migration) were encountered except for a delayed Wallstent shortening/migration at the venous anastomosis, which resulted in early access failure. The overall primary and secondary patency rates (+/- standard error) of all the vascular accesses in our 23 patients at 3, 6, 12 and 24 months were 69% +/- 9 and 88% +/- 6, 41% +/- 10 and 88% +/- 6, 30% +/- 10 and 77% +/- 10, and 12% +/- 8 and 61% +/- 13, respectively. For the forearm and upper-arm grafts, the primary and secondary patency rates were 51% +/- 16 and 86% +/- 13 vs 45% +/- 15 and 73%+/-13 at 6 months, and 25% +/- 15 and 71% +/- 17 vs 23% +/- 17 and 73% +/- 13 at 12 months (p = .346 and .224), respectively. CONCLUSION: Metallic stent placement is a safe and effective means for treating peripheral venous lesions in dialysis graft patients after insufficient balloon dilation. No statistically difference in the patency rates between the forearm and upper-arm patient groups was seen.
Vascular Patency ; Treatment Failure ; Stents ; Renal Dialysis ; Polytetrafluoroethylene ; Middle Aged ; Metals ; Male ; Humans ; Graft Occlusion, Vascular/*therapy ; Forearm ; Female ; *Arteriovenous Shunt, Surgical ; *Angioplasty, Balloon ; Aged, 80 and over ; Aged

BACKGROUNDTo reveal the characteristics of genotype and phenotype of HIV strains in blood and some tissues of AIDS patients.

METHODSThe virus was isolated from peripheral blood mononuclear cell (PBMC),cerebrospinal fluid (CSF)and lymph nodes of 3 AIDS patients by coculture with PBMC stimulated by PHA for 72 hours from uninfected donor. The cytopathic effect of the HIV isolates was determined in cultured MT2 cell line. The env gene sequences form proviral DNA were analyzed by GCG software.

RESULTSIn one patient,there were differences between the strains from blood and different tissues both in genotype and phenotype. The biological phenotypes of two strains from CSF were non syncytium (NSI) type, their env sequences were similar to standard CNS tropic strain (SF162).

CONCLUSIONSThe viral heterogeneity exists in different body compartments within an infected individual. The neurotropic isolate which is similar to international standard strain exists in some AIDS patients in China.

Acquired Immunodeficiency Syndrome ; virology ; Adult ; Coculture Techniques ; Female ; Genetic Heterogeneity ; Genotype ; HIV ; isolation & purification ; Humans ; Leukocytes, Mononuclear ; virology ; Lymph Nodes ; virology ; Male ; Phenotype

BACKGROUNDEarly detection with screening mammography can potentially reduce breast cancer mortality rates. To achieve an efficient screening, a peer review system provides a compensatory double-check reviewing, will hopefully to prevent the omission of detectable lesions and reduce unnecessary recall.

METHODSIn 2009, 4643 initial mammographic screenings reported by 74 screening radiologists had negative results with a recall rate of less than 5%. In the same year, 2538 initial positives screened by 18 screening radiologists had a recall rate higher than 15%. Those 7181 randomized screenings were evenly distributed for reassessment by 39 reviewing radiologists. The disagreement of assessments between the reviewers and screening radiologists was recorded. The differential rate was defined as the number of the disagreements divided by the number of audited films reviewed by a screening radiologist. The equality of the differential rates for each screening radiologists with negative and positive assessments was compared by a Chi-square test. The performance of the 39 auditors was measured by the Kendall's tau statistic. P values less than 0.05 were considered statistically significant.

RESULTSThe mean differential rate for screening radiologists of negative assessments was 6.7% (P = 0.588), while 35.0% for positive assessments were significant (P < 0.001). The result indicated that most of the initial negative assessments reported by the screening radiologists were generally accepted by the reviewers but not the positive assessments. With respect to the 39 reviewers, there was no significant evidence for the association of the difference rates between negative and positive assessments. Nine reviewers were found to have their differential rate for negative and positive assessments larger than the average of the population. Eleven reviewers were found to have their differential rates smaller than the average for both. Thirteen reviewers had their differential rates smaller than the average for negative assessments but larger than the average for positive assessments. The opposite condition was found for six reviewers. The Kendall's tau statistic was 0.038 (P = 0.735).

CONCLUSIONSReviewers usually agreed with the opinion of the initial screening doctors who reported negative findings. Therefore, a 5% recall rate as the lower range of reviewing negatives may be still too high. The recall rate of more than 15% was significantly related to improper interpretation, especially when the differential rate is 25% or higher, a warning to the underperforming screening radiologist is recommended. An ideal reviewer should interpret films independently. Reviewers with tendencies to be followers or contrarians should not be enrolled in the reviewing system.

Breast Neoplasms ; diagnostic imaging ; Early Detection of Cancer ; Female ; Humans ; Mammography ; Peer Review ; Reproducibility of Results ; Taiwan

OBJECTIVETo study the influence of biological characteristics on HCV/HIV co-infection, including HIV-1 sequence distance, HIV-1 viral load and CD4+ count.

METHODSHIV-1 sequence distance was calculated by Clustal W and Phylip software while HIV-1 viral load being tested by NASBA and CD4+ count was tested using Epics XL of Coulter. Significance was determined by t-test using SPSS 12.0.

RESULTSThe mean HIV-1 genetic distances were 7.95% and 15.73% (P < 0.001) between those with HCV co-infection and those without. Their mean HIV-1 viral loads were 4.61 and 4.45 (P = 0.522) and their mean CD4I T counts were 308 and 251 (P = 0.161), respectively.

CONCLUSIONData showed that in the study group, the HIV/HCV co-infection had an influence on the HIV sequence distance, but did not have major impact on HIV-1 viral load and their mean CD4+ T count.

Adult ; Blood Donors ; statistics & numerical data ; CD4-Positive T-Lymphocytes ; immunology ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; immunology ; virology ; HIV-1 ; classification ; genetics ; immunology ; Hepatitis C ; epidemiology ; immunology ; virology ; Humans ; Male ; Phylogeny

OBJECTIVEThis study was to compare the performance of three HIV antibody confirmatory assay kits in confirming early HIV infection.

METHODSFive HIV antibody-positive plasma specimens were ten-fold serially diluted and then detected by ELISA. The above diluted specimens were detected with the following three HIV antibody confirmatory assay kits to analyze their sensitivity, including Wantai-RIBA (Recombinant immunoblot assay, Beijing Wantai Biological Pharmacy, China), MP-WB (HIV Blot 2.2 WB, MP Biomedicals Asia Pacific Pte. Ltd., Singapore) and INNO-LIA (INNO-LIA(TM) HIV I/II Score, Innogenetics N.V., Belgium), respectively. These kits were further used to detect 48 ELISA-reactive specimens from 11 sets of HIV seroconversion specimens (a total of 48 samples) which were previously detected as HIV antibody-positive by ELISA.

RESULTSWhen 5 samples were diluted to 100 fold, Wantai-RIBA still can detect them positive. Among the 48 HIV antibody-positive specimens detected with ELISA, the confirmation positive rate for Wantai-RIBA, MP-WB and INNO-LIA were 97.92% (47/48), 81.25% (39/48) and 91.67% (44/48), respectively. There was statistically significant difference between the confirmatory results of Wantai-RIBA and MP-WB (χ(2) = 6.13, P < 0.05), as well as between those of INNO-LIA and MP-WB (χ(2) = 5.48, P < 0.05); however, there was no statistically significant difference between those of Wantai-RIBA and INNO-LIA (χ(2) = 1.33, P > 0.05). For other six HIV seroconversion panels containing indeterminate specimens, the average seroconversion period of time for Wantai-RIBA, MP-WB and INNO-LIA were 0.7, 13.3 and 3.7 days, respectively.

CONCLUSIONCompared with MP-WB, Wantai-RIBA and INNO-LIA could reduce the window period to confirm early HIV infection.

Early Diagnosis ; HIV Antibodies ; blood ; HIV Infections ; diagnosis ; Humans ; Reagent Kits, Diagnostic

Adolescent ; Adult ; Aged ; COVID-19/virology* ; COVID-19 Nucleic Acid Testing/methods* ; Child ; Coronavirus Nucleocapsid Proteins/genetics* ; Feces/virology* ; Female ; Humans ; Male ; Middle Aged ; Phosphoproteins/genetics* ; RNA, Viral/genetics* ; SARS-CoV-2/isolation & purification* ; Young Adult