Objective To construct and screen the human immunodeficiency virus-1 (HIV-1) negative regulation factor (Nef) peptide-specific CD4+ T lymphocyte clone.Methods Peripheral blood mononuclear cells (PBMC) from five asymptomatic HIV-1 infected patients were collected and Bulkcultured with Nef end peptides.The CD4 molecule and intracellular interferon (IFN)-gamma of cultured cells were detected by two-color flow cytometry.The Nef end peptide-specific T cell clone was then constructed by limited dilution and confirmed through enzyme linked immunospot assay (ELISPOT).The best grown cells were selected and cultured as the final clone.Results The Nef end peptide-specific-T lymphocyte clone was successfully constructed from PBMC of one HIV-infected patient and confirmed by ELISPOT.The detection of human leukocyte antigen (HLA)-DRB1 type showed that the epitope of this peptide was probably HLA-DRB1 * 0406.Conclusion The Nef end specific-T cell clone is successfully constructed,and a new epitope in the C-terminus of Nef protein and its HLA restriction are identified.

Lepidopteran heat-tolerant(ht)cell lines have been obtained with sf-9,sf-21 and several Bombyx cells.They have a distinct karyotype,membrane lipid composition,morphology and growth kinetics from the parental cell lines.In this paper,we report the development of ht cell lines from other insect species and examination of their growth characteristics and virus susceptibility.Adaptation of cell lines sf-9,BTI-TN-5131-4(High5)and BTI-TN-MG1(MG 1)to 33℃ and 35℃ was carried out by shifting the culture temperature between 28℃ and higher temperatures by a gradual stepwise increase in temperature.The process of adaption to a higher culture temperature was accomplished over a period of 2 months.The cell lines with the temperature adaption were designated as sf9-ht33,sf9-ht35,High5-ht33,High5-ht35,MG1-ht33,MG1-ht35.These cell lines have been subcultured over 70 passages.Adaption to high temperatures was confirmed by a constant population doubling time with individual cell lines.The population doubling time of heat adapted cell lines were 1-4 h less than these of parental cell lines.Cell shapes did not show obvious change,however,the cell size of sf9-ht cells was enlarged and those of High5 and MG1 ht cells were reduced after heat adaption.When the cell lines were infected with Autographa californica nuclear polyhedrosis virus(AcMNPV)at 28℃,33℃,35℃ and 37℃,production of budded virus and occlusion bodies in each cell line was optimum at its own adapted temperature.

Objective To investigate the relationship between the plasma neutrophil gelatinase-associated lipocalin (NGAL)and endothelin-1 (ET-1) levels and cognitive dysfunction in chronic obstructive pulmonary disease (COPD) patients.Methods A case-control study was performed,consisting of 128 patients with COPD(68 patients without cognitive dysfunction and 60 patients with cognitive dysfunction) and 70 normal controls.All patients with COPD were diagnosed by pulmonary function tests and plasma levels of NGAL and ET-1 were determined by enzyme immunoassay.The cognitive function was evaluated by the MMSE and MoCA.Results ①Compared with normal control,the levels of plasma NGAL and ET-1 were increased(NGAL:(2.20±0.60) μg/L vs (1.69±0.73) μg/L,P＜0.05;ET-1:(26.19± 10.55)pg/ml vs (13.05±2.37) pg/ml,P＜0.05) in COPD patients without cognitive dysfunction and in COPD patients with cognitive dysfunction(NGAL:(3.80±2.75) μg/L vs (1.69±0.73) μg/L,P＜0.01;ET-1:(37.82±0.29) pg/ml vs (13.05±2.37) pg/ml,P＜0.01).Compared with the COPD patients without cognitive dysfunction,the levels of plasma NGAL and ET-1 were also increased in COPD patients with cognitive dysfunction (all P＜0.05).②The plasma NGAL levels were correlated negatively with MMSE scores(r=-0.524,P＜0.05),and negatively correlated with MoCA scores (r=-0.527,P＜0.05).The plasma ET-1 levels were negatively correlated with MMSE scores(r=-0.549,P＜0.05),and negatively correlated with MoCA scores(r=-0.558,P＜0.05).The levels of NGAL and ET-1 were positively correlated(r=0.564,P＜0.05).Conclusion NGAL and ET-1 may be involved in the pathophysiological process of cognitive dysfunction in patients with COPD,which provides a certain clinical value for the assessment of cognitive dysfunction in patients with COPD.

Objective To investigate the inhibitory effect of small interference RNA(siRNA)on the exDression of tumor necrosis factor(TNF)-oligo nucleotide fragments were designed and synthesized according to the sequence of TNF-insert downstream to H1 promoter to construct plasmids pHS-A and pHS-B for expression of short hairpin RNA(shRNA).Then the recombinant pSilence3.1-TNF-phages(RAW264.7)by lipofectamin 2000.The inhibition of TNF-examined bv real-time PCR and ELISA,respectively.Results After LPS stimulating 6 hours,TNF-expression was increased in a specific time-dependent manner,and reached high peak at 9-12 hours.The macrophages treated with plasmids pHS-A showed a significant decrease in TNF- mRNA(0.021 34±0.009 60)and protein(149.93,P＜0.01)compared with untransfected group[TNF-mRNA:0.021 34±0.009 60,protein:1 922.30±149.05]pg/ml].The relative rate of inhibition was 83.3％.No inhibitory effects was found in plasmids pHS-B and control group.Conclusions LPS stimulation results in a increasing expression of TNF-of TNF-

Objective To investigate the alternation of insulin -like growth factor -Ⅱ( IGF-Ⅱ) gene promoter P4 methylation status in hepatocellular carcinoma ( HCC) and explore its relationship with expression of P4 mRNA levels.Methods Liver specimens of 43 patients with HCC and normal liver specimens of 9 control patients were collected in operation .Tissue DNA and total RNA were extracted from these specimens .IGF-Ⅱ P4 methylation status and P4 mRNA expression levels were detected .Results (1)The incidence of IGF -ⅡP4 methyl-ation in HCC group was significantly lower than that in normal liver specimens (16.28%vs 88.89%,χ2 =19.12,P<0.01).(2)The expression level of IGF -ⅡP4 mRNA in HCC group was significantly higher than that that in normal liver specimens[(0.96 ±0.74) vs (0.25 ±0.19),t=5.48,P<0.01].(3)In HCC group,the IGF-Ⅱ P4 mRNA expression level with hypomethylation gene was significantly higher than that without hypomethylation gene [(1.18 ± 0.76) vs (0.32 ±0.27),t=5.28,P<0.01].Conclusion The hypomethylation alternation of IGF -Ⅱ P4 gene promoter which is accomplished by up -regulate P4 mRNA expression has a close relationship with HCC .

Objective To compare intraindividual higher-order aberrations(HOA)after implantation of aspherical and spherical intraocular lenses(IOL).Design Prospective,case-controlled study.Participants 33 patients(66 eyes)were between 50 and 76 years of ages.The patients with bilateral senile cataracts were operated with phacoemulsification and IOL implantation.All the patients had no other ocular pathologies and no previous ophthalmic surgery.Patients with keratometric astigmatism higher than-2.0 diopters(D)were excluded from the study.Methods Wavefront aberration measurements of 33 patients(66 eyes)after implantation of a spherical IOL(Canon Staar KS-3)in one eye and an aspherical IOL(Cannon Staar KS-3Ai)in the contralateral eye were analyzed.The eyes were randomly assigned to groups of the spherical IOL or the aspherical IOL.Third-order,4th-order,5th-order,total HOA root-mean-squares(RMS),and 4th-order spherical aberrations(Z40)were compared 3 months after surgery.Main Outcome Measures Visual acuity(VA),the best corrected visual acuity(BCVA),the centrality of IOL,refraction data,diameter of pupil size,3rd-order,4th-order,5th-order and total HOA RMS,and 4th-order spherical aberration(Z40)RMS were measured.Results Fourth-order aberration RMS(0.193?0.098?m),Z40 RMS(0.037?0.099?m),total HOA RMS(0.498?0.072?m)in the aspherical IOL group were lower than those with the spherical IOL(0.403?0.155?m,0.381?0.142?m,0.737?0.164?m,respectively)at 6mm pupil diameter.For 3rd-and 5th-order RMS at 6mm pupil diameter,total HOA RMS(at 5mm pupil diameter),no significant difference was found in both IOLs.Under mesopic conditions,ten patients(30%)with an aspheric IOL had better contrast sensitivity.The average myopic shift were(-0.29?0.09)D and(-0.87?0.16)D in the aspherical and spherical IOL groups at 6mm pupil diameter(P

Objective To find out the relationship between kidney- yang- deficiency syndrome and the Th1/Th2 expression, and to investigate the effect of kidney- yang warming therap yon the Th1/Th2 expression. Methods Kidney- yang- deficiency rat models were firstly induced by adenine and then were sensitizated by inhalation of ovalbumin to establish allergic rhinitis (AR)models .The treatment group was given Jinkui Shenqi pill. After treatment , the changes of behavior , secretion of nasal cavity and nasal mucosa were observed. The levels of Th1 cytokines IFN- ? , IL- 2 and Th2 cytokines IL- 4 , IL- 5 in serum were determined by ELISA. Results The behavioral score in AR group and kidney- yang- deficiency AR group was higher than that in the treatment group and normal control group ( P

0.05),while loratadine group shows a highly significant treatment efficacy(P

This review focuses on biological characteristics of Echinococcus f elidis including molecular genetic markers , species status ,host coverage ,geographical distribution ,epidemiological implications ,phylogeny ,and evolution .The molecular genetic markers are involved in mitochondrial cox1 (cytochrome C oxidase subunit 1) and nad1 (nicotinamide adenine dinucle-otide dehydrogenase subunit 1) genes ,nuclear protein-coding gene sequences such as elp (ezrin-radixin-moesin-like protein) , e f1a (elongation factor 1 alpha) ,pepck (phosphoenolpyruvate carboxykinase ) ,pold (DNA polymerase delta ) ,and ribosome RNA gene sequences such as ITS1 and 18S rRNA .The establishment of species status is based on distinctly discriminated mor-phological characteristics such as hooks on the rostrum with apparent rugae ,the special definitive host (lions) ,and divergence of DNA sequences ,etc .between E . f elidis and other Echinococcus species .In brief ,the review has provided researchers and ex-perts in the field of echinococcosis with fundamental background knowledge and guidelines for future research directions ,clinical and epidemiological investigations ,and prevention and control of echinococcosis .

Objective To investigate the effect of transplantation of allogeneic bone marrow hematopoietic cells(HCs)and mesenchymal stem cells(MSCs)on experimental colitis(EC)in rats.Methods The HCs and MSCs obtained from SD male rats were cultured and expanded in vitro.In experiment 1 and 2 groups,HCs were labeled with bromodeoxyuridine(BrdU)and MSCs were obtained using the tube wall attach technique,respectively.Seventy-two female rats were infused with trinitrobenzene sulfonic acid(TNBS)to induce EC models.After 24 hours,HC or MSC suspensions were injected into the rats in experimental 1(n=18)and 2(n=18)groups via caudal veins,respectively.Control animals were injected with isotonic saline.The whole colon was removed on day 7,14 and 21 after transplantation and examined histopathologically.BrdU labeled HCs were tested with immunohistochemical staining and MSCs were detected for sex-determining gene(sry)by PCR.Results EC models were successfully established.The HCs or MSCs grew rapidly in the culture suspension.On day 7,14 and 21 after transplantation,the BrdU immunoreactive cells were detected in the colon(6/6),and the positive expression of the sry gene was found in 1/6,2/6 and 3/6,respectively.No positive labeled cell was found in controls.There was no significant improvement in histopathological scores on the colon in two experimental groups compared with the controls.Conclusions Allogeneic HCs and MSCs may localize in the colon of EC models.The ability of localization is higher in HCs than MSCs.The transplantation of HCs and MSCs can not obviously improve histopathologically.