Objective To analyze the evolution of rtI233V mutation in the reverse transcriptase domain of hepatitis B virus (HBV) and its association with adefovir dipivoxil (ADV) resistance. Methods The rate of detection of rtI233V mutation in 9830 patients with chronic HBV infection was analyzed. HBV reverse transcriptase genes isolated from serial serum samples of two patients were amplified by nested PCR, and clonal sequencing (>20 clones/sample) was performed to analyze the evolution of rtI233V mutations. The replica of pTriEx-HBV1.1 vectors harboring wild-type and mutant strains (rtI233V, rtN236T, rtI233V+rtN236T) were respectively constructed and transiently transfected into HepG2 cells. Media containing serial concentrations of lamivudine (LAM), ADV, entecavir (ETV), or tenofovir (TDF) were used to treat the cells. Then HBV DNA in the supernatants was quantitatively determined by real-time PCR in order to analyze HBV mutants' replication competence and phenotypic characteristics under the drug pressures. Results The detection rate of rtI233V mutation in 9830 nucleos(t)ide analogues-treated patients was 0.28% (28/9830), including 0.19% (19 patients) with rtI233V individual mutation and 0.09% (9 patients) with rtI233V mutation combining with rtN236T or other mutations. All of the patients had rtI233V had ADV exposure history: 16 (57.1%) of them received ADV monotherapy for over six months, and 12(42.9%) of them received ADV combined sequential therapy for over 12 months. Replication competence and phenotypic resistance analysis showed rtI233V and wild-type strains had similar viral replication competence, while rtN236T exhibited significantly lower replication competence compared with wild-type strains. rtI233V strains remained sensitive to LAM, ADV, ETV, and TDF and showed little influence on drug resistance when combined with rtN236T, but it showed ability to restore the defected replication capacity of rtN236T strains. Conclusions The rtI233V mutation is associated with sub-optimal response to ADV. Though it does not directly reduce virus sensitivity to ADV, rtI233V mutation enhances replication competence of ADV resistant strains, and it is a complementary mutation.

OBJECTIVETo investigate the cytotoxic effect of γδ T cells from osteosarcoma patients against interferon-γ (IFN-γ)-treated osteosarcoma cells in vitro.

METHODSHuman γδ T cells were amplified by zoledronate from peripheral blood cells of osteosarcoma patients. The expression of Fas on the osteosarcoma cells were measured by flow cytometry and quantitative real-time PCR analysis before and after IFN-γ treatment. The cytotoxicity of γδ T cells against osteosarcoma cells was evaluated with LDH assay.

RESULTSIFN-γ significantly enhanced the susceptibility of the osteosarcoma cell lines HOS and U2OS to the cytotoxicity of γDelta; T cells from osteosarcoma patients (P<0.01). IFN-γ obviously up-regulated the expression of Fas in HOS and U2OS cells (P<0.01). Anti-FasL mAb failed to inhibit the cytotoxicity of γδ T cells in untreated osteosarcoma targets (P>0.05), but significantly impaired γδ T cell cytotoxicity in IFN-γ pre-treated osteosarcoma targets (P<0.01).

CONCLUSIONIFN-γ can enhance the cytotoxic effect of human γδ T cells from osteosarcoma patients against osteosarcoma cells in vitro.

Bone Neoplasms ; metabolism ; Cell Line, Tumor ; Cytotoxicity, Immunologic ; Humans ; Interferon-gamma ; pharmacology ; Osteosarcoma ; immunology ; metabolism ; Receptors, Antigen, T-Cell, gamma-delta ; immunology ; T-Lymphocytes, Cytotoxic ; cytology ; drug effects ; immunology ; fas Receptor ; metabolism

Objectives To investigate the application modes of indocyanine green (ICG) angiography,digital subtraction angiography (DSA) and methylene blue angiography in the hybrid surgery of cerebral and spinal arteriovenous malformations (AVMs) and to compare the application value of different intraoperative angiographic methods.Methods From July 2013 to December 2015,55 patients treated with hybrid surgery of cerebral and spinal AVMs in Xuanwu Hospital,Capital Medical University were analyzed retrospectively,including 8 patients with cerebral AVM(Spetzler-Martin grade ≥Ⅲ) and 47 with spinal AVM.Their mean age was 33.8±15.6 years.Intraoperative DSA was performed in a hybrid operation room and methylene blue angiography was performed via the feeding artery.ICG angiography was performed in 4 cases in the initial stage as a comparison.Anatomic cure was confirmed by DSA at 3 months after surgery and the results were assessed.Results All 55 patients performed intraoperative DSA,32 underwent methylene blue angiography and 4 performed intraoperative ICG angiography.The frequency of intraoperative DSA was 3.6±1.3 times for each case.After the resection of the first lesion,the residual lesions revealed by DSA accounted for 27.3% (15/55).85.5% (47/55) patients achieved anatomic cure,in which the patients with midbrain AVM (8/8) and spinal AVM 83.0% (39/47) achieved anatomic cure.Three patients were lost to follow up at 1 year after procedure.The median modified Rankin scale (mRS) score of the cerebral AVMs was 2.0 (0.0-3.8).Compared with 3.5 (2.0-4.0) before procedure,there was significant difference (Z=-2.264,P<0.05).The Aminoff score in patients with spinal AVM after procedure was 3.5 (1.0-6.0).Compared with 4.0 (1.0-6.0) before procedure,there was no significant difference (Z=-0.262,P>0.05).The patients with function preservation (function score equal or better than pre-operation) accounted for 88.5% (46/52).Conclusions Intraoperative DSA could precisely localize the nidi and verify the complete resection.Intraoperative methylene blue angiography could selectively reveal the feeding artery supplied nidi in the operative field and identify the angioarchitecture.ICG angiography could reveal the structures of nidi on the surface of the operation area.The intraoperative methylene blue angiography could replace ICG angiography and achieve the treatment target of anatomical cure of the hybrid surgery of the cerebral and spinal cord vascular malformations.

Objective To analyze the perioperative clinical character of the severe neurological complications in intracranial aneurism treated with stent-assisted coiling(SAC). Methods 203 cases of intracranial aneurysms patients treated by SAC were enrolled retrospectively(ruptured aneurysm group 45 cases and un-ruptured,aneurysm group 158 cases)and the perioperative clinical character of the serious neurological complications(11 cases)was further analyzed. Results The total rate of serious neurological complication was 5. 4%,11 cases of patients with 13 aneurysms got 13 stents. In the ruptured aneurysm group, 5 cases(11. 1%)suffered severe neurological complications,including intraoperative bleeding in one case, postoperative stent-related ischemia in one case,both 2. 2% . Postoperative bleeding 2 cases(4. 4%),and one case of bleeding during anesthesia induced stage(2. 2%). In the unruptured aneurysm group,intraoperative bleeding in three cases,and postoperative stent-related ischemia in three cases,both 1. 9% . No bleeding case during anesthesia induced stage or postoperative period. Although active rescue treatments were performed, 8 patients eventually died,and the total mortality rate was 3. 9% . Conclusion Intracranial aneurysms patients following SAC treatment may suffer from bleeding,ischemia,severe neurological complications, severe disability,and even die. So,we have to strengthen perioperative management.

Objective To comparatively analyze the advantages and disadvantages between a double immune-diffusion assay and a rate nephelometer analysis for the detection of antigen activity of pneu -mococcal capsular polysaccharides .Methods The antigen activity of pneumococcal capsular polysaccharides of serotypes 1,6B,9V,10A,14 and 19A from four manufacturers and ATCC were analyzed by a double im-mune-diffusion assay and a rate nephelometer analysis , respectively .The effects of antiserum samples and gain values on the rate response value were evaluated .Results The sample 4 of type 9V showed no antigeni-city with a rate response value similar to that of negative control as indicated by both tests .However ,the pre-cipitation lines and the rate response values presented by other polysaccharide samples differed in a wide range.Results of the rate nephelometer analysis were not affected by the anti -serum samples from different sources and the gain values .Conclusion The rate nephelometer analysis could quantitatively analyze the antigen activity of pneumococcal capsular polysaccharides .

OBJECTIVE: To investigate the rare earth elements(REEs) contents and distribution characteristics in nasopharyngeal carcinoma( NPC) tissue in Gannan region. METHOD: Thirty patients of NPC in Gannan region were included in this study. The REEs contents were measured by tandem mass spectrometer inductively coupled plasma(ICP-MS/MS) in 30 patients, and the REEs contents and distribution were analyzed. RESULT: The average standard deviation value of REEs in lung cancer and normal lung tissues was the minimum mostly. Light REEs content was higher than the medium REEs, and medium REEs content was higher than the heavy REEs content. REEs contents changes in nasopharyngeal carcinoma were variable obviously, the absolute value of Nd, Ce, Pr, Gd and other light rare earth elements were variable widely. The degree of changes on Yb, Tb, Ho and other heavy rare earth elements were variable widely, and there was presence of Eu, Ce negative anomaly(δEu=0. 385 5, δCe= 0. 523 4). CONCLUSION The distribution characteristic of REEs contents in NPC patients is consistent with the parity distribution. With increasing atomic sequence, the content is decline wavy. Their distribution patterns were a lack of heavy REEs and enrichment of light REEs, and there was Eu , Ce negative anomaly.
Carcinoma ; Humans ; Lung ; Lung Neoplasms ; Metals, Rare Earth ; chemistry ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; chemistry ; Reference Values ; Tandem Mass Spectrometry

In order to find out the distribution of Genotype of those people infected with HBV (hepatitis B virus) from north Guangxi and the relationship between different immune status of HBV infected people and their genotypes, the HBV infected people are classified into three types according to immune tolerance, immune clearance ( response) and immune incompetence (residues). 150 cases from each type, a total of 450 cases are chosen to be tested with real time fluorescence quantitative PCR assay for detection of HBV infection in three kinds of different immune state of the HBV genotype. In the 450 cases, 323 cases belong to type B, 94 cases belong to type B, 23 cases belong to mixed type B+C and 10 cases belong to none B and none C type. Type B are the majority in all the three HBV immune status, made up to 70%, 78%, 67.33% of each type. The different immune state genotype proportion difference don't have statistical significance; immune state and genotypic correlation isn't statistically significant; type B HBV-DNA load is higher than that of type C, groups of persons aged 30 years or older with type C are significantly higher than that of < 30 years of age, the difference was statistically significant; among the genotypes of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBil) positive rate showed no significant difference between male and female; there was no significant difference in genotype distribution. The results show that, in North Guangxi HBV genotypes B, C accounts for the proportion, a small amount of B+C hybrid, occasionally fails to type HBV infection; among immune tolerance, immune clearance (response) and immune incompetence (residues) type B are in majority in these three kinds of immune state, chronic HBV infection immunity with the HBV genotype correlations were not statistically significant.
Adult ; Aged ; China ; Female ; Genotype ; Hepatitis Antibodies ; immunology ; Hepatitis B ; immunology ; virology ; Hepatitis B virus ; classification ; genetics ; immunology ; isolation & purification ; Humans ; Male ; Middle Aged ; Young Adult

Humans ; Female ; Diagnosis, Differential ; Breast/pathology* ; Carcinoma, Intraductal, Noninfiltrating/pathology* ; Breast Neoplasms/pathology*

Objective To study the distribution of hypoxia inducible factor ( HIF ) 3A gene single nucleotide polymorphisms ( SNPs ) in Guangxi Han population and compare their distribution differences with different populations. Methods We conducted SNPscan technique to detect the genotypes of rsl 1672731 and rs2072491 on 286 Guangxi Han population included in the study and statistically analyzed the genotype and allele frequency and the HapMap-CEU, HapMap- HCB, HapMap-JPT, HapMap-GIH and HapMap-MEX data differences. Results Three genotypes, AA, AG and GG, were found in rsl 1672731 of HIF3A, with frequency of 42.7%, 45. 5% and 11. 8%, respectively, the allele frequencies of A and G were 65.5% and 34.5%, respectively. Three genotypes of CC, CT and TT, were found for rs2072491 with frequency distributions of 47.6%, 43.0% and 9.4%, respectively, the allele frequencies of C and T were 69.1% and 30.9%, respectively. There was no significant differences in genotype and allele frequencies of rsl 1672731 and rs2072491 between different genders in Guangxi Han population (P>0. 05). However, compared with the typing data of CEU, HCB, JPT, GHI, TSI and MEX from human genome project (HapMap), the genotype and allele frequencies of rsl 1672731 and rs2072491 were not significantly different from those of HCB and JPT (P>0. 05). The genotype and allele frequencies of rsl 1672731 and rs2072491 were statistically different with the date of CEU, GIH, TSI and MEX published by the HapMap (P<0.05). Conclusion The polymorphisms of HIF3A gene rsl 1672731 and rs2072491 have differences on different populations.

Objective To establish a standardized quantitative enzyme-linked immunosorbent as-say ( ELISA) recommended by WHO for the detection of human IgG antibodies specific for Streptococcus pneumoniae capsular polysaccharides ( Pn PS ELISA ) .Methods According to the WHO recommended standard Pn PS ELISA protocol , capsular polysaccharide concentrations of 13 serotypes (1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 23F) of Streptococcus pneumonia for coating were optimized;the ELISA plates and AP conjugated goat anti-human IgG antibody for the detection were experimentally selected .Using the established assay parameters assured by testing quality control standards ( QCs) provided by WHO refer-ence laboratory , a panel of 16 LIBP ( Lanzhou Institute of Biological Products Co .Ltd.) QCs were measured for comparison analysis between WHO reference laboratory and LIBP laboratory .In the meantime , the range of IgG concentrations of an internal QC panel 907 for 13 serotypes of pneumococcal capsular polysaccharide were established for routine QC work in LIBP laboratory , and inter-assay precision within LIBP laboratory was assessed as well .Results The standardized Pn PS ELISA assay established in LIBP laboratory met the criteria required by WHO .There was a high correlation between the data collected by WHO reference labora -tory and LIBP laboratory (slope=0.94, coefficient of correlation r=0.97, P<0.05).Eighty one percent of IgG concentrations measured by LIBP laboratory were within the range of ±40%of those measured by WHO reference laboratory , which met the criteria of 75%data falling within the ambit of ±40%of assigned values commonly used for comparison .The ranges of IgG concentration in LIBP QC 907 for 13 serotypes had been established.The inter-assay precision in LIBP laboratory was high with coefficiency of variation ( CV) less than 30%.Conclusion LIBP laboratory has successfully established the standardized ELISA recommended by WHO for quantitative detection of human IgG antibodies against pneumococcal capsular polysaccharides (Pn PS ELISA).The range of IgG concentrations in LIBP QC 907 for 13 serotypes of pneumococcal capsular polysaccharide are also established for routine quality-control practice .