Objective Through the investigation of 80 cancer operation patients' families,we would like to investigate further more the effect of their anxiety conditions for the patients' mental state and find out the corresponding strategy.Methods 80 cancer patients' families in our department were chosen to fill the anxiety scales between the 2nd to 5th day after their admission and some corresponding directions were given.Results Compared with the norm,the cancer patints' families' anxiety scores had significant difference(P

Drugs, Chinese Herbal ; pharmacology ; Female ; Granulosa Cells ; drug effects ; secretion ; Humans

0.05).The score of GMFM-66(55.24?11.68)after treatment had obviously improved as compared with that before treatment(39.81?13.17)in the observation group(P=0),and the score of GMFM-66(48.76?11.96)after treatment had obviously improved as compared with that before treatment(40.25?12.89)in the control group(P=0.010 3),and the effect of the observation group was significantly superior to that of control group(P=0.038 0).Conclusion ESFN combined with TOT can effectively improve the motor function in children with cerebral palsy.

10.3969/j.issn.1000-3606.2013.06.015

Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P＜0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P＜0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.

Diagnostic Errors ; Female ; Humans ; Middle Aged ; Mycoses ; diagnosis ; Paranasal Sinus Diseases ; diagnosis ; Sinusitis ; diagnosis ; Tuberculosis ; diagnosis

Arrhythmias, Cardiac ; physiopathology ; Electromyography ; methods ; Female ; Humans ; Infant, Newborn ; Male

Background Krüppel-like factor 6 (KLF6) is related to the physiological or pathological process,such as growth,cell differentiation,proliferation,apoptosis,angiogenesis,tissue repair,and so on.But in ophthalmology,it is less reported about the expressing level of KLF6 protein in lens epithelial cclls (LECs) or the effect of KLF6 on the proliferation of human LECs.Objective This study was to investigate whether KLF6 can inhibit proliferation of human LECs.Methods KLF6 eukaryotic expression plasmid (pEGFP-C2-KLF6) was constructed using reverse transcription PCR(RT-PCR) and identified by double enzyme digestion method and PCR.Human LECs strain (HLE-B3) was cultured and passaged using low glucose DMEM containing 10％ fetal bovine serum and then divided into 4 groups.KLE-B3 transfection reagents were added in the culture medium of all groups.In addition,no agent was used in the blank control group;only insulin-like growth factor-1 (IGF-1) was appended to the medium in only IGF-1 group ;null vector was transfected and IGF-1 was appended in the null plasmid transfection+ IGF-1 group;while pEGFP-C2-KLF6 eukaryotic expression plasmid was transfected into the cells,and simultaneously IGF-1 was added in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group.After 24 hours of intervene,water soluble tetrazolium salt-1 (WST-1) test was used to detect the growth status of the cells,and Western blot assay was used to assay the relative expressing level of KLF6 protein in the cells.In the other hand,the cells were cultured at the density of 1 ×104/piece,and 0,0.10 and 0.25 μg pEGFP-KLF6 were transfected into each piece of cells respectively,and then IGF-1 was added with a final concentration of 50 μg/L for 24 hours after cell culture.Expressions of Ki-67 protein and mRNA in the cell pieces were detected by immunocytochemistry and fluorescent semiquantitative PCR,respectively.Results The PCR product bands were consistent with KLF-6 gene in length,and the product fragments were corresponding with expectant ones via PCR and double enzyme digestion method,showing a successful construction of pEGFP-C2-KLF6 eukaryotic expression plasmid.After 24 hours of IGF-1 stimulation,the absorbance values of the cells were 0.86±0.00,2.10±0.01,2.24±0.12 and 1.06±0.02 in the blank control group,only IGF-1 group,null plasmid transfection + IGF-1 group and the pEGFP-C2-KLF6 plasmid transfection + IGF-1 group,with a significant difference among the 4 groups (F =38.322,P ＜ 0.05),and that in the pEGFP-C2-KLF6 plasmid transfection +IGF-1 group was significantly lowed in comparison with the only IGF-1 group and the null plasmid transfection+IGF-1 group (q=6.42,7.31,both at P＜0.05).Western blot assay showed that the relative expressing levels of KLF6 protein were statistically different among the four groups (F =591.858,P＜0.05),and those in the pEGFP-C2-KLF6 plasmid transfection+IGF-1 group were 1.47,2.04,3.27 folds higher than those in the blank control group,only IGF-1 group,respectively and null plssmid transfection+IGF-1 group,respectively.Immunocytochemistry revealed that the expressing intensity of Ki-67 protein was gradually weakened with the decrease of pEGFP-C2-KLF6 plasmid dose in the 0,0.10 and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+IGF-1 groups.Fluorescence semiquantitative PCR results showed that the relative expression values of Ki-67 mRNA in the cells were O.15±0.08 and 0.11±0.03 in the 0.10 μg and 0.25 μg pEGFP-C2-KLF6 plasmid transfection+ IGF-1 groups,which were significantly lower than O.77± 0.12 of the 0 μg pEGFP-C2-KLF6 plasmid transfection group,with a statistically significant difference among the three groups (F =54.825,P＜0.05).Conclusions KLF-6 can effectively inhibit IGF-1-induced proliferation of human LECs,and it can be regarded as one of the regulatory factors of the proliferation of HLE-B3 cells.

Objective To investigate the effect of jejunal infusion of amino acids on secretion of gastrointestinal hormone in healthy dogs.Methods Six healthy adult dogs were treated with jejunal fistulas and femoral vein intubation.Twenty-four hours after the operation,solution of 8 different amino acid monomers (experimental group) or normal saline (control group) were infused into the jejunum of the dogs every 24hours.The levels of cholecystokinin (CCK),motilin,and gastrin in the peripheral plasma were measured using radioimmunoassay at the start of infusion (0 minute),and 30,60,90,and 120 minutes after infusion.Results Compared with the control group,the serum CCK level in the phenylalanine group was significantly higher 30 and 60 minutes after infusion [(1.25 ±0.19) ng/L vs.(0.66 ±0.14) ng/L,(1.23 ±0.12) ng/L vs.(0.80 ± 0.03) ng/L,both P ＜ 0.01],while that in the tryptophan group was significantly higher 30 minutes after infusion [(1.08 ±0.26) ng/L vs.(0.66 ±0.14) rig/L,P ＜0.01].The other measurement results showed no statistically significant differences.Conclusions Jejunal infusion of phenylalanine or tryptophan may stimulate the secretion of gastrointestinal hormone to some extent.Aromatic amino acids (phenylalanine and tryptophan) is more potent in triggering the release of CCK than aliphatic (leucine,isoleucine,and methionine) and charged amino acids (aspartic acid,arginine,and glutamate).The mechanism may be related to the properties of the amino acids.

OBJECTIVE:To observe therapentic efficacy of bevacizumab combined with irinotecan+leucovorin+fluorouracil (FOLFIRI)plan in the treatment of advanced colon cancer,toxic reaction and patients'survival rate. METHODS:A total of 113 patients with advanced colon cancer admitted to the oncology department in our hospital from Jan. 2010 to Aug. 2014 were random-ized into observation group 1(40 cases),observation group 2(39 cases)and control group(34 cases). Three groups received FOLFIRI;observation group 1 and 2 were additionally given Bevacizumab injection 5 and 7.5 mg/kg 14 d as a treatment course, for 8 cycles. Clinical efficaices as well as the positive rate of VEGF-A,immune indexes(the proportion of CD3+,CD3+CD4+,CD3+CD8+ in T cell subset)before and after treatment,the incidence of toxic reaction,1-year and 2-year survival rates were compared among 3 groups. RESULTS:The total response rate of observation group 1 and 2 were significantly higher than control group,and the positive rate of VEGF-A in observation group 1 and 2 were significantly lower than control group,with statistical significance (P<0.05). The proportion of CD3+,CD3+CD4+ and CD3+CD8+ in 3 groups were significantly lower than before treatment,but ob-servation group 1 and 2 were significantly higher than control group,with statistical significance(P<0.05). The incidence of grade hypertension(grade 3-4)in observation group 1 and control group were lower than observation group 2;the incidence of leucope-nia(grade 3-4)in observation group 1 and 2 were significantly lower than control group,with statistical significance(P<0.05). There was no statistical significance in 1-year survival rate among 3 groups(P>0.05). 2-year survival rate of observation group 1 and 2 were significantly higher than control group,with statistical significance(P<0.05). CONCLUSIONS:For advanced colon cancer,different doses of bevacizumab combined with FOLFIRI have significant synergistic effect,can effectively inhibit VE-FG-A,play a role of immune protection and anti-toxic side effects,and prolong the survival time. The incidence of hypertension in patients treated with low-dose bevacizumab is relatively lower and the safety is better.