In order to establish stable high expression cell lines, the eukaryotic expression vector pIRES2EGFP and recombinant plasmid pIRES2EGFP-TIM-3 were transfected into mammalian cells CHO by Lipofectamine. The transfected cells were cultivated under selective growth medium including G418 and green fluorescent protein (GFP) positive cells were sorted by FACS. Simultaneously, growing transfectants were selected only by G418 in the medium. The GFP expression in stably transfected cells was detected by FACS. Under selective growth conditions with G418, the percentage of GFP positive cells was reduced rapidly and GFP induction was low. In contrast, the percentages of GFP positive cells were increased gradually after FACS. By 3 rounds of GFP selection, the stable high expression cell lines were established. Furthermore, using FACS analysis GFP and the target protein TIM-3 co-expression in the stable transfectants cultured in nonselective medium was detected. Theses results demonstrated that the stably transfected cell lines that express high titer of recombinant protein can be simply and fleetly obtained by using GFP and selective growth medium.

Objective To investigate the incidence,correlative factors and prevention of perimenopausal symptoms.Methods 146 perimenopausal women were investigated by using Kupperman menopausal index(KMI),life event scale,improved social support scale and self-made inventory.Results We got 112 available data,the incidence of perimenopausal symptoms was 75.0%,58.0%was mild,15.2%was moderate,1.8%was sevcre.The main correlative factors of perimenopausal symptoms were inhabited environment,self emotional controllability,history of gynecologic diseases,history of depression,dysmenorrhea,premenstrual syndrome,menopause,score of life eventscale,improved social support scale.Conclusions The incidence of perimenopausal symptoms is high.It is also related to factors of society,family and mentality,except hypogonadism.Preventing perimenopausal symptoms according to its cirrelative factors may decrease its incidence and enhance the life quality of women.

Objective To analysis the effect of dezocine combined anesthesia on serum levels of IL-6,MDA and SOD in patients after laparoscopic surgery for colon cancer.Methods 48 patients who were undergoing laparoscopic radical resection of colon cancer in our hospital were collected.All patients were randomly divided into experimental group and control group, 24 cases in each group, the control group was given remifentanil combined anesthesia, the experimental group received dezocine combined anesthesia, after surgery, the serum levels of interleukin-6 (IL-6), tumor necrosis factor-α( TNF-α) , malondialdehyde ( MDA ) , superoxide dismutase ( SOD ) and incidence of adverse reactions were detected in all patients. Results After surgery, compared with control group, the serum levels of IL-6 and TNF-αwere lower in the experimental group (P<0.05);the serum level of MDA was lower, and the serum level of SOD was higher in the experimental group(P<0.05);the incidence of adverse reactions was lower in the experimental group(P<0.05).Conclusion The dezocine combined anesthesia can significantly reduce the serum levels of IL-6,TNF-αand MDA in patients after laparoscopic surgery for colon cancer,improve serum SOD level, effectively reduce inflammation and oxidative stress reaction, and have high security.

Objective To probe the effect of applying montelukast combined with pidotimod in asthma of aged patient.Methods 120 cases of asthma in non-acute episode phase were randomly divided into three groups.Group 1 was treated with montelukast combined with pidotimod,group 2 was treated with montelukast,and the control,namely group 3 was ketotifen.All cases were treated by means of intensification therapy including glucocorticoids,?_2 agonists and some necessary anti-infection during acute episode.Results After 10 weeks treatment,both group 1 and 2 made notable improvement with respect to many asthma control parameters(P

AIM: To evaluate effects of inhaled nitric oxide(iNO) on adhesion molecule CD11b expression on lung neutrophils in experimental meconium aspiration syndrome(MAS) rabbits treated with conventional mechanical ventilation under room air or 100%O 2. METHODS: Animals were randomly allocated to 8 groups( n= 48) of 6 each: two MAS model groups(under room air or 100%O 2 without iNO treatment), 6 treatment groups were treated with continuous NO inhalation at a dose of 0.2?10 -6 mol/L,0.33?10 -6 mol/L or 0.67?10 -6 mol/L respectively for 12 hours under room air or 100%O 2. Mean systemic arterial pressure(SAP) and methemoglobin (MeHb) were performed at basement time, 0, 2, 4, 12 hours. Expression of CD11b on neutrophils in the bronchoalveolar lavage fluid(BALF) was detected with flow cytometry. RESULTS: SAP, MeHb at different time among different groups were within the normal scale. CD11b expression on the neutrophils in the BALF significantly decreased in groups of inhalation 0.33?10 -6 mol/L or 0.67?10 -6 mol/L NO, compared with the two MAS model groups. ( ?s : under 21%O 2, 0.33?10 -6 mol/L NO,121?20 vs 392?204; 0.67?10 -6 mol/L NO,112?30 vs 392?204;under 100%O 2,0.33?10 -6 mol/L NO,113?24 vs 293?65; 0.67?10 -6 mol/L 102?114 vs 293?65, P 0 05). No statistic difference was observed between groups inhaled 0.33?10 -6 mol/L NO and 0.67?10 -6 mol/L NO. CONCLUSION: 0.33?10 -6 mol/L or 0.67?10 -6 mol/L NO inhalation down-regulated the CD11b expression on the neutrophils in BALF to reduce the sequestration of neutrophils in rabbit lung.

Objective To summarize the nursing experience of 1 case of very low birth weight infant decannulation difficult in PICC. Methods The nursing key points included: full assessment analysis decannulation difficult reason, consult the PICC catheter outpatient health nurses, give magnesium sulfate hydropathic compress, mucopolysaccharide polysulfate cream local besmear outside, sanyrene outside, at the same time give low-molecular-weight heparin calcium injection subcutaneous injection such as anticoagulant active treatment and nursing. Results After 7 days ,the infant left axillary mass dispel, PICC pull out smoothly. Conclusions Decannulation difficult of very low birth weight infant requires full evaluation, multidisciplinary cooperation and specialist consultation, can give targeted personalized nursing safety smooth tube drawing, is worthy of reference for clinic.

The effects of drugs on intracellular calcium concentration(［Ca2+］i) were investigated with fura-2 fluorescence technique to investigate ATP and thrombin-induced Ca2+ entry in bovine aortic endothelial cells(BAEC). It was found that application of ATP and thrombin gave rise to biphasic ［Ca2+］i elevation. ATP or thrombin only triggered a fraction of cyclopiazonic acid(CPA)-sensitive Ca2+ store, which was enough to activate Ca2+ entry. The Ca2+ release induced by thrombin resulted from the activation of phospholipase C(PLC), whereas the PLC-independent mechanism was involved in ATP-induced Ca2+ release. Nifedipine had no effect on ATP and thrombin- induced Ca2+ entry. SK&F 96365 and ginsenoside-2A inhibited both ATP and CPA-induced Ca2+ entry, however no effect of them on thrombin-induced Ca2+ entry was found. The inhibitory effects of SK&F 96365 and ginsenoside-2A on CPA-induced Ca2+ entry were less than that on ATP-induced Ca2+ entry. The Ca2+ influx sensitive to SK&F 96365 was not the same as that to ginsenoside-2A. These observations suggest that both ATP and thrombin evoke Ca2+ release and Ca2+ influx by activation of different receptor. However their mechanisms appear different.

Objective To compare the severity of periopemtive myocardial injury caused by live donorliver transplantation (LDLT) performed under sevoflurane and pmpofol combined anesthesia in adult patients. Methods Forty ASA Ⅱ-Ⅳ patients(liver function Child-Pugh grade B or C)aged 40-67 yr weighing 47-95 kgundergoing right lobe LDLT were randomly divided into 2 groups(n=20 each):sevoflurane combined anesthesiagroup (group S) and propofol combined anesthesia group(group P).Anesthesia was maintained with 1.6%-3.0% sevoflurane in group S or PCI of pmpofol(Cp 2-4μg/ml)in group P combined with sufentanil infusion at 0.5- 1.0 μg·kg-1·h-1 and intermittent iv boluses of cisatracurium in both groups.MAP,HR,CVP,MPAP,PCWP, CO and mixed venous O2 saturation(S(v)O2)were recorded before skin incision at 5 and 30 min of anhepatic phaseand 5 and 30 min of ncohepatic phase and the end of operation:Blood samples were taken from central vein beforeskin incision(T0,baseline)at 30 min of anhepatic phase(T1),30 min of neohepatic phase(T2),the end ofsurgery (T3) and 24,48,72 h after operation (T4,5,6) for determination of serum concentrations of cardiactroponin I (cTnI) and creatine kinase MB(CK-MB).Postoperative adverse effects were recorded.Results Thetwo groups were comparable with respect to 8ex ratio(M/F),age,body weight,duration of operation,duration ofanhepatic phase,blood loss,dopamine and nitroglycerin consumption.There was no significant difference in MAP, HR,CVP,MPAP,PCWP,CO and S(v)O2 between the two groups.The serum concentrations of cTnI and CK-MB were significantly increased at T2-5 as compared with the baseline value at T0 in both groups. There was no significant difference in serum concentrations of cTnI and CK-MB and in the incidence of myocardial ischemia and arrythmia between the two groups. Conclusion Sevoflurane and propofol combined anesthesia have similar effects on myocardial injury caused by LDLT in adult patients.

Objective To investigate the clinical and pathological characteristics of IgA nephropathy (IgAN) in association with active tubular interstitial lesions. Methods 148 patients who were diagnosed as IgAN by renal biopsy and admitted to Zhejiang Provincial People's Hospital from March 2014 to December 2014 were enrolled. They were divided into IgAN with active tubular interstitial lesions group (IgAN?ATIL group, 23 patients) and IgAN without active tubular interstitial lesions group (control group, 125 patients). Clinical and pathological characteristics were retrospectively analyzed. Multivariate logistic regression analysis was used to analyze the influence factors. Results The prevalence of ATIL in 148 IgAN patients was 15.5%. IgAN?ATIL group showed an older average age and more higher proportion of medication history (antibiotics, diuretics, nonsteroidal anti ?inflammatory drugs, etc) than those in control group. There were significant differences in Alb, eGFR, Scr, BUN, 24?hour urinary protein quantity, urinary NGAL and urinary RBC count between two groups (P<0.05, respectively). A moderate of tubulointerstitial lesions of IgAN?ATIL group was shown, while the control group was mainly mild lesions. Multivariate logistic regression analysis showed that age, medication history and the urinary NGAL level were independent risk factors of IgAN ? ATIL. Conclusions IgAN patients with active tubular interstitial lesions had more severe clinical manifestations and chronic interstitial lesions. The age, medication history (antibiotics, diuretics, nonsteroidal anti?inflammatory drugs, etc) and the urinary NGAL level were independent risk factors of IgAN?ATIL.

Objective To explore the quality of life and the related factors in adult epilepsy patients in rural areas of Gansu province. Methods The Quality of Life in the Epilepsy-31 Inventory Chinese version (QOLIE-31) were used to evaluate the quality of life in 154 epilepsy patients and 149 healthy controls. The relationship of demographic variables (gender, age, marital status, profession and education, etc.) and clinical variables (age of onset, duration, type of seizures, seizure frequency and medication, etc.) with the quality of life were analyzed in patients. Results Compared with the con?trol group, the QOLIE-31 score was significantly lower in epileptic group [(47.63±7.74) vs. (52.28±5.75), P<0.05]. There was no significant difference in the patients' QOLIE-31 score between difference ages, genders, marital status, occupa?tions, and educations (P>0.05). The differences of patients' QOLIE-31 score were significantly different between differ?ent incomes, medication types of epilepsy (P<0.05). The patient's family income (β=3.115, P=0.002) and the type of medi?cation (β=3.261, P=0.027) were factors affecting their quality of life. Conclusion The quality of life in adult patients with epilepsy is lower. Economic status of family and the number of drug type significantly affects the quality of life in patients. Reasonable drug choice to treat seizure and reduce the economic burden of the family are effective measures to improve the quality of life of patients with epilepsy.