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1.
Article in English | WPRIM | ID: wpr-878357

ABSTRACT

Objective@#The aim of the present study was to evaluate the performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one dried blood spot (DBS) as an alternative sample to plasma.@*Method@#A total of 571 paired DBS/plasma samples were collected from men who have sex with men (MSM) and injection drug users (IDUs), and serological and molecular assays were performed. Using plasma results as the reference standard, the performance of DBS tests for HIV-1 RNA, HIV-1 DNA, and HCV RNA was evaluated. Pearson's correlation coefficients and Bland-Altman analysis were performed to assess the correlation and concordance between DBS and plasma.@*Results@#Among paired plasma/DBS samples with detectable HIV-1 RNA and HCV RNA, five samples (5/32) were not detectable in DBS, while measurable HIV-1 RNA levels were present in plasma (1.44 to 3.99 log @*Conclusion@#The performance of the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA using one DBS was acceptable. DBS, as an alternative sample to plasma, may be a viable option for the simultaneous detection of HIV-1 RNA, HIV-1 DNA, and HCV RNA in resource-limited settings or for individuals living in areas that are difficult to access.


Subject(s)
DNA, Viral/analysis , Diagnostic Tests, Routine/methods , Dried Blood Spot Testing/methods , HIV Infections/diagnosis , HIV-1/isolation & purification , Hepacivirus/isolation & purification , Hepatitis C/diagnosis , RNA, Viral/analysis , Sensitivity and Specificity , Specimen Handling/methods , Syphilis/diagnosis , Treponema pallidum/isolation & purification
2.
China Journal of Endoscopy ; (12): 100-104, 2018.
Article in Chinese | WPRIM | ID: wpr-702915

ABSTRACT

Objective To study the etiology clinical features, treatment outcomes and prevention of esophageal submucosal hematoma caused by endoscopic injection sclerotherapy (EIS) of esophageal varices. Methods We retrospectively reviewed the clinical data of patients who were diagnosed with esophageal submucosal hematoma caused by EIS and treated in our hospital from Jan 2014 to July 2016. Five patients were analyzed including one patient receiving endoscopic gastrointestinal catheterization combined with medication, and the remaining four received medication therapy only. Results All five patients were discharged with clinical improvement. However the patients treated only with medication therapy recovered more slowly than the ones who treated with combined therapy. No treatment related side-effects were observed among two treatment groups. Conclusion Endoscopic gastrointestinal catheterization combined with medication may be an effective treatment for esophageal submocasal hematoma caused by EIS. However, the actual clinical efficacy and safety remain to be proven by future large sample randomized clinical studies.

3.
Article in English | WPRIM | ID: wpr-264574

ABSTRACT

A real-time RT-PCR (RT-qPCR) assay for the detection of Tahyna virus was developed to monitor Tahyna virus infection in field-collected vector mosquito samples. The targets selected for the assay were S segment sequences encoding the nucleocapsid protein from the Tahyna virus. Primers and probes were selected in conserved regions by aligning genetic sequences from various Tahyna virus strains available from GenBank. The sensitivity of the RT-qPCR approach was compared to that of a standard plaque assay in BHK cells. RT-qPCR assay can detect 4.8 PFU of titrated Tahyna virus. Assay specificities were determined by testing a battery of arboviruses, including representative strains of Tahyna virus and other arthropod-borne viruses from China. Seven strains of Tahyna virus were confirmed as positive; the other seven species of arboviruses could not be detected by RT-qPCR. Additionally, the assay was used to detect Tahyna viral RNA in pooled mosquito samples. The RT-qPCR assay detected Tahyna virus in a sensitive, specific, and rapid manner; these findings support the use of the assay in viral surveillance.


Subject(s)
Animals , Culicidae , Virology , Encephalitis Virus, California , Real-Time Polymerase Chain Reaction , Methods , Reverse Transcriptase Polymerase Chain Reaction , Methods , Sensitivity and Specificity
4.
Article in Chinese | WPRIM | ID: wpr-231154

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of human cytomegalovirus (HCMV) on the cell cycle of duct epithelial cell cultures of human salivary gland (HSG) in vitro and relative mechanism.</p><p><b>METHODS</b>HSG was cultured in vitro. Reverse transcriptase polymerase chain reaction (RT-PCR) and nest-RT-PCR were used respectively to investigate ie1/ie2 transcription in HSG infected by human cytomegalovirus(HCMV). The effects of HCMV on the cell cycle of HSG were studied by flow cytometry in vitro. The expression of cyclin D1 in HSG infected by HCMV was detected by Western blotting.</p><p><b>RESULTS</b>HCMV iel/ie2 transcription could be detected in HSG infected by HCMV. HCMV arrested productively infected cells in G1 stage. And cyclin D1 was down-regulated in HCMV infected HSG.</p><p><b>CONCLUSION</b>HCMV inhibits proliferation of HSG by affecting G1/S check point and down-regulating cyclin D1 in vitro.</p>


Subject(s)
Blotting, Western , Cell Culture Techniques , Cell Cycle , Physiology , Cyclin D1 , Genetics , Metabolism , Cytomegalovirus , Physiology , Epithelial Cells , Cell Biology , Virology , Flow Cytometry , Humans , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Salivary Glands , Cell Biology , Metabolism
5.
Article in Chinese | WPRIM | ID: wpr-237610

ABSTRACT

This study was aimed to explore the effects of peptidoglycan (PGN) on proliferation and cell cycle of human bone marrow mesenchymal stem cells (MSCs). MSCs were isolated from human bone marrow by density gradient centrifugation. The purity of MSCs with the spindle fibroblastic morphology was identified by microphotography and the phenotypes were detected by flow cytometry (FCM). MSCs incubated with different doses of PGN (1, 10, 20 μg/ml) were used as test groups, and those incubated without PGN were regarded as control group. The isolated and cultured MSCs were inoculated into 96-well plates according to a certain concentration. Cell cycle was measured by flow cytometry after incubated with PGN for 72 hours. The results showed that the cell proliferation index was significantly increased in dose and time dependent manners after MSCs was incubated with PGN. Its effects on the proliferation of MSCs were highest in 10 μg/ml group. Compared with the control group, PGN could significantly decrease proportion of MSCs in G₀/G₁ phase and increase them in S and G₂/M phases (p < 0.05). It is concluded that PGN can promote more MSCs to enter the DNA synthesis phase and proliferate many much MSCs in dose and time dependent manners.


Subject(s)
Bone Marrow Cells , Cell Biology , Cell Cycle , Cell Proliferation , Cells, Cultured , Flow Cytometry , Humans , Mesenchymal Stem Cells , Cell Biology , Peptidoglycan , Pharmacology , Toll-Like Receptor 2
6.
Article in Chinese | WPRIM | ID: wpr-334043

ABSTRACT

The aim of this study was to explore the characteristics of Toll-like receptor expression in mesenchymal stem cells derived from bone marrow of healthy donor (BM-MSCs). BM-MSCs were isolated from bone marrow of healthy donor by Ficoll method. Expressions of CD34, CD45, HLA-DR, CD44 and CD71 in BM-MSCs were detected by flow cytometry. CD71 in BM-MSCs was assayed by immunocytochemistry. The adipocyte and osteoblast induction of BM-MSCs were detected by alizarin red stain and oil red stain respectively. TLR 1 - 10 mRNA levels in BM-MSCs were evaluated by semiquantitative RT-PCR. The results showed that expressions of CD34, CD45 and HLA-DR in BM-MSC were negative while the expressions of CD44 and CD71 were positive. CD71 in BM-MSCs was positive. After induced by osteoblast and adipocyte inductor, BM-MSCs were positive for alizarin red staining and oil red staining respectively. All of TLR 1 - 10 mRNA were found in BM-MSCs with high expression levels of TLR2, TLR3, TLR4, TLR7, TLR8, TLR9 and low expression levels of TLR1, TLR5, TLR6, TLR10. In conclusion, different levels of TLR 1 - 10 mRNA were expressed in BM-MSCs of healthy donor.


Subject(s)
Bone Marrow Cells , Metabolism , Cell Differentiation , Cells, Cultured , Humans , Mesenchymal Stem Cells , Metabolism , RNA, Messenger , Genetics , Toll-Like Receptors , Metabolism
7.
Acta Pharmaceutica Sinica ; (12): 563-566, 2002.
Article in Chinese | WPRIM | ID: wpr-251100

ABSTRACT

<p><b>AIM</b>To study the effects of various liposomes formulations and preparation methods on the stability of acyclovir palmitate (ACV-C16) liposomes on storage at 4 degrees C and 25 degrees C over a 6 months period.</p><p><b>METHODS</b>The mean particle size, Zeta potential, pH and leaking ratio of ACV-C16 liposomes were the parameters chosen to indicate the stability of liposomes. All of the parameters were compared among various lipid compositions [egg lecithin/cholesterol/hosphatidylserine (PC/CH/PS), egg lecithin/cholesterol/stearylamine (PC/CH/SA), egg lecithin/cholesterol/cholesteryl sulphate (PC/CH/CS), bovine brain ceramides/cholesterol/palmitic acid/cholesteryl sulphate (CM/CH/PA/CS)], different preparation methods (film dispersing, reverse phase evaporation, dehydration/rehydration), charges (positive, negative), as well as among multilamellar vesicles liposomes (MLV), large unilamellar vesicles liposomes (LUV) and dehydration/rehydration vesicles liposomes (DRV).</p><p><b>RESULTS</b>An analysis of various parameters led to the conclusion that the stability of liposomes followed the order of PC/CH/CS > CM/CH/PA/CS > PC/CH/PS > PC/CH/SA at the same storage conditions; the positively charged system showed the most unstable delivery system of liposomes as compared to the other three systems. As far as stability was concerned, LUV liposomes proved to be superior to MLV liposomes and DRV liposomes, and the modified reverse phase evaporation method of Szoka provided the best preparation method. The stability in systems was enhanced when systems were stored at 4 degrees C as compared to storage at 25 degrees C.</p><p><b>CONCLUSION</b>The stability of liposomes was significantly interrelated with lipid composition of various liposomes, preparation method and different storage conditions.</p>


Subject(s)
Acyclovir , Chemistry, Pharmaceutical , Drug Carriers , Drug Delivery Systems , Drug Stability , Liposomes , Chemistry , Palmitates , Chemistry , Particle Size , Technology, Pharmaceutical , Methods
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