Objective To construct the eukaryotic expression vector of linker for activation of T cells(LAT) gene in mouse. MethodsLAT cDNA of mouse mononuclear cells was amplified with polymerase chain reaction.The fragment was inserted into the eukaryotic expression vector of pCMV-HA plasmid.The recombinant plasmid was verified by DNA sequencing and used to transfect lymphocytes of the asthmatic model. Results The length of amplified fragment was 729 bp.The sequence of LAT gene was confirmed by Genbank.The LAT protein could be detected in the lymphocytes of asthmatic model.Conclusion The recombinant eukaryotic expression vector pCMV-HA-LAT can be successfully constructed.

Objective The purpose of this study was to explore the effect of nursing intervention on xerostomia in patient with AIDS. Methods From October 2010 to April 2012, nursing interventions were administered in 53 AIDS patients with xerostomia during treatment. The self-designed evaluation form was used to evaluate the oral dryness of patients before and after nursing intervention. Results There were significant changes on oral dryness of patients before and after nursing invention ( < 0.01) . There was a progressive decrease in the occurrence degree of oral dryness from level 0 to level 4. The number of patients decreased by 11.4%in level 4, but increased by 9.4% in level 0. There was a positive correlation between the intervention time and treatment effect. Establishment of an effective oral care hygiene behavior intervention could help patients create a healthy lifestyle. Conclusion Nursing intervention is important to relieving xerostomia of AIDS patients.

Objective To investigate the changes and significance of concentration of soluble intercellular adhesion molecule-1(sICAM-1) in cerebrospinal fluid (CSF) of bacterial meningitis(BM) in rabbits. Methods A total of 36 rabbits were randomly divided into m eningitis group, meningitis cefotaxime-treated group and control group. BM indu ced by escherichia coli(Ec) via cerebellomedullary cistern inoculated. Normal sa line was injected in control group. CSF was sampled in different time. The conce ntration of CSF sICAM-1 was detected by ELISA.Results 1.There was a low concentration of sICAM-1 in CSF in 85 percent of normal rabbi ts.2.In meningitis group, there was a sharp rise in the concentration of CSF sIC AM-1 at 6 hours after Ec was inoculated, reached a peak level at 12 hours, and t here was higher concentration of CSF sICAM-1 between 6 and 24 hour than that at 0 hour.3.In meningitis cefotaxime-treated group, the concentration of CSF sICAM -1 at 6 hour and 12 hours was similar to meningitis group, the time that get pe ak level was at 24 hours that at 12 hours after making use of antimicrobial agen t.The peak level was higher than meningitis group. The concentration of CSF sICA M-1 decreased markedly at 48 hours that made use of antimicrobial agent 36 hour s later,but the concentration was still higher than the peak level of meningitis group.Afterwards, with the time of making use of antimicrobial agent lengthened ,the concentration of CSF sICAM-1 decreased gradually.4.Experimental results in dicated by correlating analysis to these data that there was positive correlatio n in the concentration of CSF sICAM-1 with the brain water content.Conclusions sICAM-1 participates in the pathological process of BM, and contributes to the damage of blood brain barrier and the formation of brain edema.There is importan t significance that drugs which can resist the discharge of sICAM-1 will be impl ied. J Appl Clin Pediatr,2005,20(2):163-165

Objective:To analyze the polymorphism in human cDNA sequence of prothymosin-?(ProT?)by sequencing analysis.Methods:The cDNA of human ProT? was amplified from cells of peripheral blood and cord blood by RT-PCR.The product of RT-PCR was purified and linked with vector pMD18-T.After cloning and sequencing,the sequence of ProT? cDNA was compared with the standard sequence to analyze the polymorphism in the ProT? cDNA sequence.Results:The cloned ProT? cDNA sequence was different from that of the standard.We found 2 kinds of variations:(1)The nucleotide in 107 position was varied and the nucleotides in 110-121 and 191-205 positions were deleted;(2)The nucleotide in 306 position was deleted,mainly in the 60-80 years old group.Conclusion:We have identified 2 kinds of variations in human ProT? cDNA,but the first 28 amino acid in the N-terminal of cDNA of human ProT? are not involved therefore the variations do not affect the function of human ProT?.

ObjectiveTo investigate the correlation between serum C-reactive protein(CRP),tumor necrosis factor-alpha(TNF-α) and atherosclerosis in patients with subclinical hypothyroidism.Methods Seventy-five patients with subclinical hypothyroidism were divided into two groups according to the level of TSH:mild group (42 cases) with TSH between 5.5 and 10.0 mU/L and severe group (33 cases) with TSH ＞10.0 mU/L.Thirty healthy physical examinees or volunteers were selected as control group.TG,LDL-C,HDL-C,TSH,FT4,FT3,CRP,TNF-α and carotid intima-media thickness(CIMT) of all cases were detected and measured.ResultsCIMT of mild,severe and control groups were(0.88 ± 0.20),( 1.12 ±0.21 ),(0.62 ±0.21 ) mm,respectively.CIMT of mild and severe groups were both significantly higher than that of control group,CIMT of severe group was significantly higher than that of mild group (P ＜ 0.01 ).There were no significant differences of CRP and TNF- α among three groups (P ＞ 0.05 ).LDL-C of mild and severe groups were significantly higher than that of control group [ (3.22 ± 0.37),(3.49 ± 0.38 ) mmol/L vs.(2.48 ±0.41 )mmol/L,P ＜0.01 ].HDL-C and TG between mild and control groups had no statistical significance (P ＞0.05).Compared with control and mild groups,HDL-C of severe group decreased [(0.92 ±0.10)mmol/L vs.( 1.21 ± 0.14),( 1.17 ± 0.11 ) mmol/L] and TG increased [ ( 1.50 ± 0.49) mmol/L vs.( 1.11 ±0.53),(1.27 ±0.47) mmol/L](P＜ 0.01 or ＜ 0.05).The levels of TG,LDL-C,TSH,CRP and TNF-α had positive correlation with CIMT (r =0.52,0.37,0.48,0.39,0.45;P ＜0.05 or ＜0.01 ).FT4 was negatively correlated with CIMT (r=-0.24,P＜0.05).HDL-C had no relation with CIMT (r =0.06,P ＞0.05).ConclusionsThere are many abnormalities such as metabolism disorder of blood lipid in patients with subclinical hypothyroidism.The risk of atherosclerosis increases in these patients and inflammatory reaction may not be an important factor of subclinical hypothyroidism complicated with atherosclerosis.

0.05).The serum PICP of tubercular meningitis children after 4 neeks glucocorticoid therapy (108.85?46.13) ?g/L was significantly lower than that in control group((154.38)?47.98) ?g/L and glucocorticoid- pretreatment (152.99?44.78) ?g/L (P

OBJECTIVETo observe the clinical effect of electric stimulation of long-term retaining needle on analgesia after laparoscopic cholecystectomy (LC) and the impacts on the post-surgical flatus time.

METHODSUnder static absorptive composite general anesthesia, 90 cases of LC were randomized into three groups, 30 cases in each one. In the control group, the analgesia was not applied after LC. In the analgesia-pumper group, the patient controlled intravenous analgesia (PCIA) was used. In the needle-retaining group, the electric acupuncture stimulator was used. The needles were inserted transversely at Riyue (GB 24), Qichong (ST 30) and Yanglingquan (GB 34) and fixed with sterile sticker. Separately, in 8 h and 24 h after surgery, the electric acupuncture stimulation with disperse-dense wave, 2 Hz/100 Hz frequency was applied continuously for 30 min. Visual analogue scale (VAS), adverse reactions such as vomiting and nausea and the postoperative flatus time in 2, 4, 8, 12, 24 and 36 h after surgery were observed and recorded in the three groups.

RESULTSIn 2, 4, 8, 12 and 24 h after surgery, VAS scores in the needle-retaining group and the analgesia-pumper group were all lower than those in the control group (P < 0.05, P < 0.01). The analgesia effect at the above time points in the needle-retaining group was better than that in the analgesia-pumper group (all P < 0.05). There was not adverse reaction in the needle-retaining group. But there were 3 cases of somnolence, 6 cases of nausea and 3 cases of vomiting in the analgesia-pumper group, and 2 cases of nausea and 1 case of vomiting in the control group. The flatus time was quite earlier in the needle-retaining group as compared with the other two groups [(14.77 +/- 4.99) h vs (18.50 +/- 4.22) h, P < 0.01; (14.77 +/- 4.99) h vs (18.17 +/- 4.69) h, P < 0.05].

CONCLUSIONThe electric stimulation of long-term retaining needle is safe and effective in analgesia after LC. It avoids the adverse reactions of analgesics and promotes postoperative flatus.

Acupuncture Analgesia ; instrumentation ; Adult ; Aged ; Cholecystectomy, Laparoscopic ; adverse effects ; Electroacupuncture ; instrumentation ; Female ; Humans ; Male ; Middle Aged ; Pain Management ; Pain, Postoperative ; etiology ; therapy

Central Nervous System Infections ; cerebrospinal fluid ; Child ; Child, Preschool ; Humans ; Infant ; Insulin-Like Growth Factor Binding Protein 3 ; cerebrospinal fluid ; Insulin-Like Growth Factor II ; cerebrospinal fluid ; Male

Objective To investigate the effects of chronic hyperglycemia on the proliferation,secretion function and expression of endoplasmic reticulum stress(ERS) related molecules in pancreatic ? cell line MIN6. Methods MIN6 cells were treated with different concentrations of glucose(5.6,25 and 33.3mmol/L) and were harvested at indicated time(24h and 96h) for examinations.Cell proliferation was tested using CCK-8 solution,insulin and proinsulin secretion function was determined by ELISA,and mRNA expression of ERS related molecules(Total XBP-1,Spliced XBP-1) and protein expression of phosphorylated IRE1? were detected by Real-time PCR and Western blotting,respectively. Results After treatment with hyperglycemia for 96h, cell proliferation was significantly lower than that treated for 24h(P

BACKGROUNDAllergic asthma is associated with airway inflammation and hyperresponsiveness caused by dysregulated production of cytokines secreted by allergen-specific helper T-type 2 (Th2) cells. The linker for activation of T cells (LAT) is a membrane-associated adaptor protein, which has been shown to take part in regulating T cell receptor (TCR) signaling and T cell homeostasis. In this study, we established an asthmatic mouse model to examine the changes in LAT levels during allergic airway disease and the effects of LAT transgenic expression on airway inflammation.

METHODST cells from mouse lung tissues were isolated from allergen challenged (ovalbumin (OVA)) and control mice, and the purity of these isolated T cells was examined by fluorescence-activated cell sorter (FACS). Semi-quantitative RT-PCR and Western blotting were used to detect the expression of the LAT gene and LAT protein, respectively. After an intranasally administered mixture of pCMV-HA-LAT plasmid and Lipofectamine 2000, 24 hours before and 72 hours after allergen challenge, the BALF cell count and the differential cytologies were studied. In addition, IL-4 and IFN-γ levels in the BALF were determined by ELISA, and pathological changes in lung tissues were observed.

RESULTSLAT protein and mRNA expression were decreased in lung T cells in a mouse model of allergen-induced airway disease. After intranasal administration of pCMV-HA-LAT, histopathological examination of the lungs showed that intervention with LAT overexpression prevented mice from developing airway inflammation, and the number of total cells, eosinophils, neutrophils, and lymphocytes in the BALF was reduced significantly compared with the OVA sensitized and challenged group. In addition, the Th2 cytokine IL-4 decreased, while the Th1 cytokine IFN-γ increased compared to the OVA sensitized and challenged group or the OVA sensitized group plus pCMV-HA treatment.

CONCLUSIONThis study demonstrates that LAT might effectively diminish Th2 cytokine responses, lung histopathological changes and lung inflammation to allergen challenge in a model of experimentally induced asthma.

Animals ; Asthma ; immunology ; metabolism ; Blotting, Western ; Bronchoalveolar Lavage Fluid ; immunology ; Cells, Cultured ; Cytokines ; metabolism ; Female ; Inflammation ; immunology ; Mice ; Mice, Inbred BALB C ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; metabolism