OBJECTIVE To define the regularity of survival ability of HIV in natural environment,and prevent(infection) through contacting with positive body fluids during daily life or medical work.METHODS Having been diluted by sterile water or 10% serum RPMI 1640 medium,HIV was exposed to 4℃,room temperature(20-26℃) or 37℃ for different period of time.TCID_(50) of these samples was detected.Non-pathological samples were blind passaged for three generations.RESULTS HIV infective ability persisted more than 35 days both in(water) and medium at 4℃;whereas it persisted 7-14 days in water,14-21 days in medium at room temperature and 37℃.CONCLUSIONS HIV has higher resistance in natural environment.To prevent accidental spreading of HIV,HIV positive liquids and contaminants staffs should be treated carefully.

The mediastinal lymph node tuberculous abscesses (MLNTAs) are secondary to mediastinal tuberculous lymphadenitis.Surgical excision is often required when cold abscesses form.This study was aimed to examine video-assisted thoracoscopic surgery (VATS) for the treatment of MLNTA.Clinical data of 16 MLNTA patients who were treated in our hospital between December 1,2013 and December 1,2015 were retrospectively analyzed.All of the patients underwent the radical debridement and drainage of abscesses,and intrathoracic lesions were removed by VATS.They were also administered the intensified anti-tuberculosis treatment (ATT),and engaged in normal physical activity and follow-up for 3 to 6 months.The results showed that VATS was successfully attempted in all of the 16 MLNTA patients and they all had good recovery.Two patients developed complications after surgery,with one patient developing recurrent laryngeal nerve injury,and the other reporting poor wound healing.It was concluded that VATS is easy to perform,and safe,and has high rates of success and relatively few side-effects when used to treat MLNTA.

OBJECTIVEFrequency, type and clinical implications on protease and reverse transcriptase drug resistance mutations were investigated and phylogenetic analysis in antiretroviral drug-naïve AIDS patients was carried out in Henan province.

METHODS45 plasma samples were separated from the anticoagulatory whole blood, from which reverse transcription-polymerase chain reaction technique was used to amplify the partial pol gene. The sequences were analysed for genotypic antiretroviral resistance and phylogenetic relation through landing the websites http://hivdb.stanford.edu and http://hiv-web.lanl.gov, under BioEdit and DNAClub software.

RESULTSPartial pol sequences of 36 samples were successfully amplified. The major mutation rate of resistance to protease was 8.3% (3/36), including types D30A, V32A, G73C and V82A. Minor mutation rate of resistance was 100%, including types of L63PS (36/36), I93L (35/36), V77IL (34/36), A71IVT (10/36) and D60E (2/36). The mutation rate of resistance to reverse transcriptase was 38.9% (14/36). Mutation-scoring and clinical implication clewed drug resistance rates were 5.6% (2/36) and 22.2% (8/36) to protease inhibitors and reverse transcriptase inhibitors respectively, while 1 sample was potentially low-level resistant to all of the protease inhibitors and 3 samples to part of the reverse transcriptase inhibitors. Phylogenetic analysis revealed that the pol gene of 36 samples were highly homologous and having a near relative to B.US.83.RF ACC M17451. 36 samples seemed to have the same infection source while their resistance mutations were not due to drug-resistant virus infection but to the evolving of virus in vivo.

CONCLUSIONMost of the antiretroviral drug-naïve AIDS patients in Henan province were sensitive to the currently available antiviral medicine, but antiviral treatment must be in accordance with the strict procedure and to keep better adherence, to avoid the epidemics caused by drug-resistant virus.

Acquired Immunodeficiency Syndrome ; genetics ; Adult ; Anti-HIV Agents ; pharmacology ; China ; Drug Resistance, Viral ; genetics ; Female ; Genes, pol ; genetics ; Genotype ; HIV Protease ; genetics ; HIV Protease Inhibitors ; pharmacology ; Humans ; Male ; Mutation ; Phylogeny ; RNA-Directed DNA Polymerase ; genetics ; Reverse Transcriptase Inhibitors ; pharmacology

OBJECTIVETo comparatively investigate the inorganic composition and crystallographic properties of cortical and cancellous bone via thermal treatment under 700 °C.

METHODSThermogravimetric measurement, infrared spectrometer, X-ray diffraction, chemical analysis and X-ray photo-electron spectrometer were used to test the physical and chemical properties of cortical and cancellous bone at room temperature 250 °C, 450 °C, and 650 °C, respectively.

RESULTSThe process of heat treatment induced an extension in the a-lattice parameter and changes of the c-lattice parameter, and an increase in the crystallinity reflecting lattice rearrangement after release of lattice carbonate and possible lattice water. The mineral content in cortical and cancellous bone was 73.2wt% and 71.5wt%, respectively. For cortical bone, the weight loss was 6.7% at the temperature from 60 °C to 250 °C, 17.4% from 250 °C to 450 °C, and 2.7% from 450 °C to 700 °C. While the weight loss for the cancellous bone was 5.8%, 19.9%, and 2.8 % at each temperature range, the Ca/P ratio of cortical bone was 1.69 which is higher than the 1.67 of stoichiometric HA due to the B-type CO₃²⁻ substitution in apatite lattice. The Ca/P ratio of cancellous bone was lower than 1.67, suggesting the presence of more calcium deficient apatite.

CONCLUSIONThe collagen fibers of cortical bone were arrayed more orderly than those of cancellous bone, while their mineralized fibers ollkded similar. The minerals in both cortical and cancellous bone are composed of poorly crystallized nano-size apatite crystals with lattice carbonate and possible lattice water. The process of heat treatment induces a change of the lattice parameter, resulting in lattice rearrangement after the release of lattice carbonate and lattice water and causing an increase in crystal size and crystallinity. This finding is helpful for future biomaterial design, preparation and application.

Animals ; Bone Density ; physiology ; Bone and Bones ; chemistry ; ultrastructure ; Crystallography ; Spectrophotometry, Infrared ; Swine

OBJECTIVEUndried silver-hydroxyapatite-titania (Ag-nHA-nTiO2) nanoparticles slurry was used to make membrane with polyamide 66 (PA66) by co-polymerization method. The purpose of this study is to test the physical and chemical characteristics and antibacterial ability.

METHODSThe morphology, chemical components and structures of the membrane were characterized by atomic absorption spectrometer (AAS), X-ray diffraction (XRD), scanning electron microscope (SEM) and energy-dispersive X-ray analysis (EDX). Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Porphyromonas gingivalis (P. gingivalis), Fusobacterium nucleatum (F. nucleatum) and Streptococcus mutans (S. mutans) were utilized to test the antibacterial effect.

RESULTSXRD results demonstrated that the membrane have characteristic diffraction peaks of pure hydroxyapatite (HA). A homogeneous distribution of the Ca, P, Ti and Ag element in the membrane was confirmed by EDX. Both surface and section showed porous structure which was confirmed by SEM and the average hole size was 20-30 microm. The bacteria assay reflected to the antibacterial effect, 50.10% of S. aureus and 56.31% of E. coli were killed. However, 91.84% of P. gingivalis, 90.64% of F. nucleatum and 90.49% of S. mutans were killed and pictures of SEM showed obviously fewer cells on the surface.

CONCLUSIONThe nanocomposite membrane could be one of the bioactive materials with antibacterial properties for oral guided bone regeneration technique.

Anti-Bacterial Agents ; Bone Regeneration ; Durapatite ; Escherichia coli ; Nanocomposites ; Nylons ; Silver ; Staphylococcus aureus ; Titanium ; X-Ray Diffraction

OBJECTIVETo evaluate the effects of the nano-hydroxyapatite/polyamide 6 (n-HA/PA6) on the proliferation and osteoblastic differentiation of rat bone marrow mesenchymal stem cells (BMSCs), and the feasibility of using both for constructing tissue engineered bone in the calvarias of rats with critical sized defects.

METHODSThe third passage of BMSCs were cultured in osteoblastic medium and seeded on the scaffolds of n-HA/PA6, the proliferation of the BMSCs was tested by MTT (3-{4,5-dimethylthiazol-2yl}-2,5-diphenyl-2H-tetrazolium-bromide) on scheduled dates, and the osteoblastic differentiation of the BMSCs were measured by alkaline phosphatase (ALP) staining. Furthermore, the scaffolds with or without BMSCs in rat calvarial defects, after 4 weeks, 8 weeks, and 16 weeks have been implanted. Histology and scanning electron microscope were used to test the bone healing in the different groups.

RESULTSThe BMSCs seeded on the n-HA/PA6 grew well, the proliferation of cells was not affected by the scaffold, and the staining of ALP was also positive. At 4 week and 8 week after implantation, the n-HA/PA6 with BMSCs showed more new bone formation on the surface of scaffolds, with a better osseointegration of implant and host bone when compared with the group of n-HA/PA6 without BMSCs. However, there was no significant difference between these two groups at 16 week.

CONCLUSIONThe porous n-HA/PA6 has no negative effects on the proliferation and osteoblastic differentiation of rat BMSCs, and using BMSCs as seed cells and n-HA/PA6 as scaffolds is a good choice for constructing tissue engineered bone due to the enhanced new bone formation and osseointegration.

Animals ; Bone Marrow Cells ; Bone and Bones ; Caprolactam ; analogs & derivatives ; Cell Differentiation ; Durapatite ; Mesenchymal Stromal Cells ; Polymers ; Rats ; Tissue Engineering ; Tissue Scaffolds

OBJECTIVETo clone, identify and phylogenetically characterize a clade B-Thai HIV isolate representing the most prevalent virus in Henan province.

METHODSPeripheral blood mononuclear cells (PBMCs) from an HIV-1 infected patient in Henan Province were separated, and co-cultivated with phytohemagglutinin-stimulated healthy donor PBMCs. Proviral DNA was extracted from productively infected PBMCs. The full-length HIV-1 genome was amplified by using the LA Tag long template PCR system. Primers were positioned in conserved regions within the HIV-1 long terminal repeats. Purified PCR products were T-A ligated into a pWSK29-T vector(CNHN 24 clone). Three recombinant clones containing virtually full-length HIV-1 genome were identified by PCR. The full-length genome was sequenced by using the primer-walking approach. Nucleotide sequence similarities were calculated by the local-homology algorithm. Phylogenetic trees of gag, pol and env reading frames were constructed using the Phylip software.

RESULTSHIV-1 C3V4 sequences indicate that the epidemic in this area was B-Thai subtype. V3 loop multiple amino acid sequence alignments showed amino acid alterations at nine positions. The 9,010 bp genomic sequence derived from isolate CNHN 24 contained all known structural and regulatory genes of an HIV-1 genome. No major deletions, insertions, or rearrangements were found. The highest homologies of the gag, pol, vpr, and vif reading frames to the corresponding clade B-Thai RL 42 sequences were 95.42%-97.08%. Phylogenetic trees showed the closest relationship of CNHN 24 and RL 42.

CONCLUSIONThe cloning and characterization of a virtually full-length HIV-1 B-Thai subtype in central China was completed in our laboratory. The data should be helpful to future studies on the genetic diversity of HIV-1.

Amino Acid Sequence ; Base Sequence ; Blood Donors ; China ; Cloning, Molecular ; DNA, Viral ; genetics ; Female ; Genome, Viral ; HIV Infections ; virology ; HIV-1 ; classification ; genetics ; Humans ; Leukocytes, Mononuclear ; virology ; Phylogeny ; Reading Frames ; Sequence Analysis, DNA ; Sequence Homology

Objective:To investigate the subtype distribution of HIV-1 strains prevalent in four areas of China,and to study the characteristics of gag gene variation and changes in antigen epitopes under the host immune pressures. Methods:The plasma of HIV-1 infected people from Henan, Guangdong, Sichuan and Beijing in China were collected. Virion RNA was extracted directly from plasma after the virion was condensed. The gag gene was amplified by RT-PCR and nested-PCR.Sequences were subtyped by Genotyping Tool software, and phylogenetic analysis of gag gene were performed using the MEGA 4.1 software.The gene distances intra each subtype were calculated by Distance program. The Ks/Ka ratios were calculated using SNAP program. The variation analysis of CTL antigen epitopes restricted by main HLA-Ⅰ specificities in China was performed.Results:Six subtypes or circulating recombinant forms(CRFs)of HIV-1,including B',CRF07_BC,CRF01_AE,B,CRF08_BC and CRF02_AG,were identified in four areas of China.The gene distances intra each subtype were CRF01_AE>B>CRF08_BC> CRF07_BC>B' listed in order of size, meanwhile the order of Ks/Ka ratios was CRF01_AE>B>CRF08_BC>B'>CRF07_BC. Far more diversity of antigen epitopes in P17 region was observed than that in P24.Epitope mutations intra subtypes were CRF01_AE>B>B'>CRF07_BC listed in order of size. Conclusion:Itseems that CRF01_AE is under the strongest immune pressures,and displays the most diversity of gene and variation of epitopes intra subtypes prevalent in China, followed by subtype B, B' and CRF07_BC. The discrepancy of epitope mutations intra the subtypes is significant.

OBJECTIVETo investigate the FDG uptake characteristics, the factors affecting 18F-FDG uptake and the extra CT diagnostic value of 18F-FDG PET/CT scan in the malignant tumor with false negative 18F-FDG PET image.

METHODSThe data of PET/CT image in 17 patients with various kinds of cancers were reviewed and analyzed by visual observation and semi-quantity analysis ( SUV). The results were compared with the CT and histopathological diagnosis, respectively.

RESULTSOf 6 well-differentiated HCC patients confirmed by histopathological diagnosis, one had two lesions in the right lobe of the liver. One of these two lesions showed low FDG uptake on 18F-FDG PET scan and low density on CT scan. The other one was not shown on either 18F-FDG PET or plain CT scan. But on enhanced CT scan, these two lesions were found to be inhomogeneous with high density at arterial phase. The false negative 18F-FDG PET images of one gastric signet ring cell carcinoma in the gastric fundus with right adnexa metastasis, 3 renal cell carcinoma, one greater omentum and peritoneal metastatic adenocarcinoma and one well-differentiated prostate cancer were caused by normal physical uptake in the digestive tract or FDG retention in the urinary system due to normal excretion. The size of three metastases was smaller than or equal to 1 cm in diameter, however, two primary lesions of these metastases showed high FDG uptake and only one was negative on either 18F-FDG PET or CT scan. In this series, 68.8% of the primary tumors and 66.7% of metastases were found to show abnormal density on CT scan, and 31. 2% of the primary tumors and 33. 3% of metastases were not detectable on either PET or CT images.

CONCLUSIONFalse negative 18F-FDG PET in malignant tumor may be correlated with the pathologic type, differentiation degree and the lesion size. Combining CT information with PET or paying attention to the scan methods during 8 F-FDG PET examination may reduce the rate of false negative 18F-FDG PET diagnosis in various kinds of malignant tumors.

Adenocarcinoma ; diagnostic imaging ; secondary ; Adult ; Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; pathology ; Carcinoma, Renal Cell ; diagnostic imaging ; pathology ; Carcinoma, Signet Ring Cell ; diagnostic imaging ; pathology ; False Negative Reactions ; Female ; Fluorodeoxyglucose F18 ; Humans ; Kidney Neoplasms ; diagnostic imaging ; pathology ; Liver Neoplasms ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Omentum ; diagnostic imaging ; Peritoneal Neoplasms ; diagnostic imaging ; secondary ; Positron-Emission Tomography ; methods ; Radiographic Image Enhancement ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Tomography, X-Ray Computed

BACKGROUNDIt is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20% - 30%. The aim of this study was to evaluate allele-specific real-time PCR (ASPCR) to detect the resistance-related mutations located at positions 103, 184 and 215.

METHODSWe developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140 clinical samples using this method.

RESULTSThe sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M184I, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained.

CONCLUSIONSDrug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Y emerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.

Alleles ; Drug Resistance, Viral ; HIV-1 ; drug effects ; genetics ; Humans ; Mutation ; Real-Time Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity