2.The association between HLA-A antigen carrying Bw4 epitope and HIV-1 viral loads and CD+4 T cell counts
Jianping CHEN ; Mingming JIA ; Hongwei LIU ; Sha LIU ; Xiaoqing ZHANG ; Hongring ZHAO ; Yiming SHAO ; Kunxue HONG
Chinese Journal of Laboratory Medicine 2009;32(4):383-387
Objective To investigate the impact of HLA-A antigens carrying Bw4 epitopes on disease course of HIV-1 infection.Methods Three hundred and forty subjects chronically infected with HIV-I were recruited and their HLA-A and B alleles were genotyped with sequence-based high resolution typing assay.HLA-Bw genotypes of these HIV-1 infected subjects were determined and their association with CD+4 T cell counts and viral load were analysed.Results When compared with subjects canting no Bw4 epitopes in HLA-A and HLA-B loci (OBw4) (median of CD+4 T cell counts:294/μ1;plasma viral load median 6.29×104 copies/ml),CD+4 T cell counts in subjects with genotypes of 1Bw4-A and 2Bw4-AA were comparable (307 and 308/μ1,respectively),but higher viral load (1.53×105 and 2.68×105 copies/ml,respectively) was observed.In subjects with Bw4 epitopes in HLA-B alleles but no in HLA-A,significantly higher CD+4 T cell counts (417/μ1,P=0.013) and lower viral load (2.10×104 copies/ml,P=0.007) were observed compared with those without Bw4 in HLA-A and HLA-B.Conclusion HLA-B antigens carrying Bw4 epitope were protective in HIV-1 infection by maintaining higher CD+4 T cell counts and lower viral load,but such protective effect was not observed in HLA-A antigens earring Bw4 epitope.
3.Clinical applicability of xTAG liquidchip technology as EGFR mutations detection method in lung cancer
Yuanjie GUO ; Wei WANG ; Sha FU ; Fang LIU ; Jing GUO ; Jianyong SHAO
Chinese Journal of Clinical and Experimental Pathology 2014;(11):1242-1246
Purpose To analyze EGFR exon 18 ~21 gene mutations in lung cancer, and compare xTAG liquidchip technology and Sanger sequencing technology in clinical practice. Methods 1 139 tumor tissue samples from phaseⅠtoⅣlung cancer patients were randomly collected. DNA was extracted from the samples. EGFR gene mutation status in exon 18~21 was detected by xTAG liquidchip technology and Sanger sequencing technology respectively. Results The mutation status of EGFR was obtained by xTAG liquidchip technology in 1 134 patients, and 1 105 by Sanger sequencing technology, detection success rate was 99. 56% and 97. 01% respective-ly. The sensitivity and specificity of xTAG liquidchip technology Comparing with Sanger sequencing was 99. 59% and 94. 54%. Sever-al cases of multiple mutations were detected by both methods. All mutation types detected by two methods are fully consistent. Conclu-sions Comparing with Sanger sequencing technology, xTAG liquidchip technology, which is able to detect mutations of exon 18~21 simultaneously, is more convenient and efficient for EGFR gene mutation detection in lung cancer.
4.Prenatal diagnosis of sirenomelia with 3D skeletal ultrasound mode imaging, 3D spiral CT and MRI
Sha, HE ; Xia, ZHU ; Xiaohong, YANG ; Sheng, ZHAO ; HuiJing, MA ; Jianbo, SHAO ; Xinlin, CHEN
Chinese Journal of Medical Ultrasound (Electronic Edition) 2014;(10):804-808
ObjectiveTo explore the clinical value of three-dimensional (3D) skeletal ultrasound mode imaging, 3D helical computer tomography (3D-HCT) and MRI in diagnosing lower limb skeletal malformations of fetal sirenomelia.MethodsSeven fetuses were suspected of sirenomelia with routine prenatal ultrasonography examination. Three-dimensional skeletal ultrasound mode imaging and MRI were used to conifrm the diagnosis, and the results were compared to those of pathology, 3D spiral CT or X-ray after termination. Five of them underwent chromosome examination including chorionic villus or umbilical cord biopsy.ResultsSix fetuses were singletons and one fetus was a conjoined twin. Three fetuses were male, while four fetuses were female. All fetuses with sirenomelia showed varying degrees of skeletal abnormalities: 1 case of typeⅢ, 2 cases of typeⅣ, 3 cases of typeⅤ and 1 case of typeⅥ. No foot was detected in one case and only single foot was detected in other 6 cases. In 7 cases, 3D skeletal ultrasound mode imaging could demonstrate all the lower limb skeletal malformations, including abnormal femur and tibioifbula, single foot or no feet. Prenatal MRI could demonstrate abnormal femur in 4 cases, abnormal tibioifbula in 1case, and no foot malformation. The results of 3D spiral CT after termination were consistent with X-ray and pathological examination results.ConclusionsAs a new imaging technology for detecting fetal skeletal malformations, prenatal 3D skeletal ultrasound mode imaging and postnatal 3D spiral CT both can display fetal bone clearly. They both have important clinical value in diagnosing lower limb skeletal malformations.
5.Optimization, validation and application of an assay for the activity of HMG-CoA reductase in vitro by LC-MS/MS
Jing WANG ; Jiye SUN ; Chunjie SHA ; Yufeng SHAO ; Yanhong LIU ; Youxin LI ; Zhenwen DUAN ; Wanhui LIU
Journal of Pharmaceutical Analysis 2015;(6):383-388
A stable HMG-CoA reductase (HMGR) reaction in vitro was developed by a sensitive, selective and precise liquid chromatography–tandem mass spectrometry (LC–MS/MS) method. The optimized enzyme reac-tion condition contained 1.5μg of HMGR, 20 nM of NADPH with 50 min of reaction time. The method was validated by several intra-and inter-day assays. The production transitions of m/z 147.0/59.1 and m/z 154.0/59.1 were used to detect and quantify mevalonolactone (MVAL) and MVAL-D7, respectively. The accuracy and precision of the method were evaluated over the concentration range of 0.005–1.000μg/mL for MVAL and 0.010–0.500μg/mL for lovastatin acid in three validation batch runs. The lower limit of quantitation was found to be 0.005μg/mL for MVAL and 0.010μg/mL for lovastatin acid. Intra-day and inter-day precision ranged from 0.95%to 2.39%and 2.26%to 3.38%for MVAL, 1.46%to 3.89%and 0.57% to 5.10% for lovastatin acid, respectively. The results showed that the active ingredients in Xuezhikang capsules were 12.2 and 14.5 mg/g, respectively. This assay method could be successfully applied to the quality control study of Xuezhikang capsule for the first time.
6.The Targeted-deletion Technology in the Monascus ruber Mediated via Agrobacterium tumefaciens
Yan-Chun SHAO ; Li LI ; Sha YANG ; Ying ZHAO ; Xiao-Hong WANG ; Fu-Sheng CHEN ;
Microbiology 1992;0(02):-
Monascus spp.,a kind of filamentous fungi,produce abundant of important metabolites which were widely used in the fields of food and medicine.Until now,there are few reports on the important functional genes of the Monascus spp.due to little genetic information.In this paper,the feasibility of gene deletion mediated via Agrobacterium tumefaciens on the basis of homologous recombination was analyzed by studying on the deletion of the RGS domain of putative G-protein signaling regulator gene mrfA in Monascus ruber.The length of homologous arms of deletion vector pC805S were 958 bp and 824 bp,respectively.There were 26 transformants in which homologous recombination occurred in 138 transformants and the recombination rate was 18.8%.The result showed it was feasible to identify the function of unknown gene in M.ruber with the targeted-deletion technology mediated via A.tumefaciens.
7.Analysis of Nef-specific interferon-γ secretion responses in HIV-1 B/C recombinant virus infectors
Hongwei LIU ; Kunxue HONG ; Jun MA ; Lin YUAN ; Sha LIU ; Jianping CHEN ; Yuanzhi ZHANG ; Yuhua RUAN ; Zhe WANG ; Yiming SHAO
Chinese Journal of Laboratory Medicine 2009;32(7):754-759
Objective To analyze characteristics of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype B/C infectors. Methods 19 HIV-1 recombinant subtype B/C infectors infected within 1 year, 40 chronic infectors infected for more than 3 years were enrolled in this cohort study. Elispot assay was used to observe HIV-1 specific T lymphocyte responses in HIV-1 recombinant subtype B/C infectors. Results Nef-specific T lymphocyte responses of interferon-gamma secretion were identified in 15 Chinese HIV-1 recombinant subtype B/C infectors infected within 1 year. The specific T lymphocytes were mainly targeted at four peptides which span from Nef 83 to 135: EVA7081.1, EVAT081.5, EVA7081.6 and EVAT081.48. Responses were identified in 29(75. 2%) infectors with more than 3 years of infection and the specific T lymphocytes were mainly targeted at three peptides which span from Nef 63 to 101 : EVA7081.43, EVA7081.44, EVAT081.45, EVA7081.47, EVA7081.48 and EVA7081.49. The average magnitude of response in infectors with less than 1 year of infection was 284. 13 SFC/106 PBMC. The average magnitude of response in infectors with more than 3 years of infection was 152. 44 SFC/106 PBMC. There was a significant difference between the two groups (U = 91. 000, P = 0. 002). Conclusions HIV-1recombinant subtype B/C infectors at different stages of diseases (less than 1 year and more thank 3 years) can recognize central region of Nef. The magnitude of Nef-specific IFN-γ secretion T lymphocyte responses in this cohort gradually decrease with disease progression.
8.Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype C/B' and subtype B' in-fectors
Hongwei LIU ; Kunxue HONG ; Jun MA ; Lin YUAN ; Sha LIU ; Jianping CHEN ; Yuanzhi ZHANG ; Yuhua RUAN ; Zhe WANG ; Yiming SHAO
Chinese Journal of Microbiology and Immunology 2008;28(11):998-1003
Objective To analyze character of Nef-specific T lymphocyte responses in Chinese HIV-1 recombinant subtype C/B' and subtype B' infectors and to identify the common immunodominant re-gions recognized by these infectors. Methods Fifty-nine HIV-1 recombinant subtype C/B' infectors and 27 subtype B' infectors were tested by IFN-γ enzyme linked immunospot (ELISPOT) assay using HIV-1 C/B' Nef overlapping peptides. Results Nef-specific T-cell responses of IFN-γ secretion were identified in 44 (74.58%) out of 59 HIV-1 recombinant subtype C/B' infectors. Ten peptides, EVA7081.1,5, 6, 7,43, 44, 45, 47, 48, 49 were mainly recognized. Amino acid position was from Nef63 to 115 and 117 to 139. Twenty of 27 (74.07%) HIV-1 subtype B' infectors recognized peptides. EVA7081.1,2, 43 and 49 were mainly recognized. Amino acid position was from Nef 63 to 77 and 87 to 119. There was no correlation be-tween the Nef-specific IFN-production of HIV-1-specific T cells responses and viral load or CD4 T cell count in both subtype infectors. Conclusion The immunodominant regions, from Nef63 to 77 and 87 to 115 were recognized by both Chinese HIV-1 recombinant subtype C/B' infectors and subtype B' infectors. These re-gions could be used in design of vaccine.
9.Simvastatin protect retinal ganglion cells against optic nerve crush in mice
Xuan ZHANG ; Zhen Wen AN ; Bo Shao DU ; Kang LI ; Li Xue SHA ; Lan GAO
Recent Advances in Ophthalmology 2017;37(12):1105-1109
Objectivc To investigate the protective effect of simvastatin on retinal ganglion cells (RGC) after optic nerve crush (ONC).Methods Together 50 Kunming mice were randomly divided into normal group,sham group,ONC group and simvastatin protection group.The mice in the normal group were untreated,the sham group was treated with the exposure of the optic nerve without injury,the ONC group mice underwent ONC operation on the left eyes,followed by intravitreal administration of equilibrium solvent [50 mg · mL-1 ethanol plus 1 mol · L-1 NaOH,which were activated by 1 mol · L-1 HC1 (pH 7.2)],and the simvastatin protection group was intravitreally injected different concentrations of simvastatin (0.5 g · L-1,1.0 g · L-1,1.5 g · L-1) after ONC operation.Brn3a immunofluorescence staining,HE staining and toluidine blue staining were used to detect the apoptosis of RGC and pathological changes of optic nerve.Results On day 7 after operation,in the ONC group,the apoptosis of RGC was observed obviously,with the survival rate dropping to (35.1 ± 3.9) %,and the thickness from the retinal ganglion cell layer to outer nuclear layer was decreased from (123.13 ± 1.04) μm to (97.48 ± 2.33) μm,which was significantly thinner than that in the control group (P < 0.01);moreover,the fibrous bundle of optic nerve disappeared,and the neuroglial cells were clustered into groups,as well as the axons showed swelling and serious degeneration,but after intravitreal injection of 1.0 g · L-1 simvastatin,the survival rate of retinal ganglion cells increased to (76.3 ± 3.7) % (P < 0.05),and the aforementioned thickness was increased to (111.39 ± 4.06) μm,which was statistically significant when compared with the ONC group (P < 0.01).The degeneration of optic nerve was improved,and the structure of neuroglial cell axons and the nerve fibers became normal.Conclusion Simvastatin can reduce the optic nerve degeneration and improve the survival rote of retinal ganglion cells.
10.Effect on therapeutic effect of inducing the formation of the post-moxibustion sore for bronchial asthma.
Jian-Mei SHA ; Xiao-Juan DENG ; Zhi-Chi SHAO
Chinese Acupuncture & Moxibustion 2012;32(4):305-308
OBJECTIVETo observe the impact of inducing the formation of the post-moxibustion sore on the efficacy of bronchial asthma treated with scarring moxibustion.
METHODSThree hundred and seventy-two cases diag nosed definitely as bronchial asthma at remission stage were randomly divided into a modified nursing group (248 cases) and a conventional nursing group (124 cases). The scarring moxibustion was applied at Dazhui (GV 14), Feishu (BL 13), Danzhong (CV 17) and Tiantu (CV 22) in either group. The direct moxibustion with moxa cone was adopted. In modified nursing group, 0.5% Iodine was used for the sterilization at moxa abscess. The herbal plaster was cut into an inverted triangle and compressed on the wound. After suppuration, the fester was not cleaned in each dressing change. Additionally, the patients were advised to have high-protein diets after moxibus tion till abscess dropped. In conventional nursing group, 0.5% Iodine was used for the sterilization at moxa abscess. The herbal plaster was cut into a round shape that could cover completely the moxa wound. After suppuration, in each dressing change, the wound was cleaned and sterilized. Additionally, the patients were advised to avoid any stimulating food after moxibustion till abscess dropped. The clinical efficacy, the change of C3 content in blood serum and the clinical symptom score were observed in two groups.
RESULTSThe total effective rate was 93.5% (232/248) in modified nursing group, which was better than 79.0% (98/124) in conventional nursing group (P < 0.01). C3 content increased apparently after treatment as compared with that before treatment in either group (both P < 0.01), but C3 level increased much more apparently in modified nursing group as compared with that in conventional nursing group (P < 0.01). The clinical symptom score reduced apparently after treatment in either group (both P < 0.01), but that reduced much more apparently in modified nursing group as compared with that in conventional nursing group (P < 0.01).
CONCLUSIONIn the treatment of bronchial asthma with scarring moxibustion, to induce the formation of the post-moxibustion sore achieves the better clinical efficacy as compared with conventional nursing.
Acupuncture Points ; Adolescent ; Adult ; Aged ; Asthma ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult