AIM:To supply the theoretic evidences of elongating the lifetime of neuron-like cells differentiated from adult rat mesenchymal stem cells, we investigated the relationship between the differentiation and apoptosis in the process of induction. METHODS: The mesenchymal stem cells(MSCs) were isolated primarily from rat bone marrow, and purified by passage culture. The 5th passage of MSCs was induced by ?-mercaptoethanol and all-trans-retinoic acid (ATRA). After 1 h, 3 h and 5 h of induction, the cells were stained immunocytochemically with anti-MAP-2 and anti-GFAP antibodies, respectively. In addition to counting the ratio of neuron-like cells in MSCs, DAPI staining was employed to identify whether the differentiated cells have an apoptotic morphological changes. The ratio of apoptotic cells at 1 h, 3 h and 5 h after induction were detected by flow cytometry (FCM). RESULTS:Neuron-like cells appeared among MSCs and could be identified by MAP-2 staining 30 min after MSCs being treated by ?-mercaptoethanol and ATRA. However, 5 h after induction, some of these neuron-like cells began to float and die. With DAPI staining, the nuclei of some dying cells were shown to be condensed and later fragmented. The ratios of cells undergoing apoptosis at 1 h,3 h and 5 h after induction by ?-mercaptoethanol and ATRA were 1.5%?6.1%?21.5% and 1.5%?1.7%?12.3%, respectively . CONCLUSIONS:1. ?-mercaptoethanol and ATRA had the different ability that induced MSCs to differentiate to neuron-like cells. 2. Apoptosis was also initiated in the process of differentiation, and there is positive correlation between the ratio of differentiation and apoptosis.

Objective In previous experiment we proved that rat marrow stromal cells(MSCs) could be induced to differentiate to neuron\|like cells by ? mercaptoethanol and all\|trans\|retinoic acid(ATRA), but the neuron\|like cells showed apoptosis sooner after the differentiation.In this study, we used several sets of combinations of anti\|apoptotic reagents and inductive reagents to induce the differentiation of MSCs,to prevent the differentiated cells from apoptosis and to elongate the lifetime of the neuron\|like cells which might be use in clinic. Methods The MSCs were isolated primarily from bone marrow of adult rat,and purified by passage culture.The 5th passage of MSCs was induced by single or combinative induction,which included RA,Melatonin(10 -9 mol/L), bFGF,bFGF+RA,Melatonin+RA,Melatonin+bFGF.At 1?h,5?h and 24?h of induction, the ratio of neuron\|like cells and the numbers of survived neuron\|like cells among 6 groups were compared.In addition,immunocytochemical stainings with anti\|MAP\|2 and GFAP were also performed on cells of 5?h of induction. Results Neuron\|like cells were all MAP\|2 positive,but GFAP negative.The ratios of neuron\|like cells in MSCs at 1?h after induced by six combinations of reagents were 0 24,0 28,0,0,0 44,0 64;at 5?h were 0 56,0 28,0,0,0 76,0 83 and at 24?h were 0 10,0 27,0,0 14,0 35,0 59 respectively.The ratios of survived neuron\|like cells of differentiated cells at 24?h after induction by RA,Melatonin,Melatonin+RA and Melatonin+bFGF were 0 08,0 37,0 05,0 51 respectively.Conclusions\ 1.bFGF can't induce MSCs into neuron\|like cells independently.Moreover,it suppressed the differentiation effects of RA,while enhancing the effects of MT.2.MT can induce MSCs differentiating into neuron\|like cells and has the anti\|apoptotic effects on the induced neuron\|like cells.3.When MT and bFGF were added together,not only the ratio of neuron\|like cell in MSCs was increased, the lifetime of neuron\|like cells was also elongated.

BACKGROUND:To evaluate the psychological characteristics of the patients waiting for renal transplantation in order to analyze coping style, thus perform the corresponding personalized clinical psychological intervention which is conductive for patients to face reality and disease. OBJECTIVE:To investigate the psychological characteristics of the patients waiting for renal transplantation and to analyze the effect of different social factors on the coping style of patients and to explore the corresponding clinical mental intervention method METHODS:Fifty-eight patients waiting for renal transplantation and 60 patients without renal transplantation were randomly selected from Second Department of Urology, Organ transplantation Center, the 309th Hospital of PLA between February 2009 and August 2010, the patients were required to fil in a psychological questionnaire form using the currently general used medical response. The effect of social factors on the coping style and psychological stress status was analyzed with multi-factor Logistic regression analysis. RESULTS AND CONCLUSION:From the questionnaire survey, the evasive and yielding value scores of the patients waiting for renal transplantation were significantly higher than those of the patients without renal transplantation (P<0.05), but there was no significant difference of confrontation score between the patients (P>0.05). It indicated that the psychological pressure of the patients waiting for renal transplantation was bigger that of the patients with out renal transplantation, which may related with course of the disease, cost and the rejection after transplantation for the patients has to endured. Multi-factor Logistic regression analysis showed that sex, education, income, payment methods and age of the patients were al related with the confrontation, evasive and yielding scores, as those with technical secondary school education or better got the best confrontation score while those with annual income less than 60 000 yuan got the highest score in evasive and yielding styles. The results show that income and education level of patients are the main factors that affect the coping style and psychological stress status of the patients waiting for renal transplantation, treating the patients with reasonable renal clinical care interventions can help the patients face to the disease positively and can reduce the damage of transplantation to the psychology of the patients.

Objective To investigate and establish thyroid function reference range during 3 pregnant stages in Chaoshan area:thyrotropin (TSH),free thyroxin (FT4).Methods 863 healthy pregnant women visited antenatal clinic in the Second Affiliated Hospital were included in this study.By detecting serum levels of TSH,FT4,thyroglobulin antibody (TgAb),and thyroid peroxi-dase antibody (TPOAb)in these women,the euthyroid reference ranges of serum TSH and FT4 in different pregnant stages were established based on the standards of the United States National Institute of Clinical Biochemical (NACB)guidelines.Results The ranges of serum TSH in the first,second and third trimester of pregnancy were 0.13-3.93 mIU/L,0.14-4.55mlU/L,and 0.42-3.85 mIU/L,respectively.The ranges of serum FT4 levels in these 3 stages were 12.00-23.34 pmol/L,6.27-12.65pmol/ L, and 9.80-18.20 pmol/L,respectively.The range of serum TSH levels in our study were similar with the reference range recom-mended by the guidelines in China,but were different compared with the American Thyroid Association (ATA)guidelines.Conclu-sion TSH,FT4 levels show dynamic changes during different stages of pregnancy.The thyroid function ranges of healthy pregnant women in Chaoshan area are quite different compared with non-pregnant population and pregnant women in distinct area.Due to the differences of diet,iodine intake,economy and culture in Chaoshan area,regional thyroid function reference ranges specific for differ-ent stages of pregnant women should be established.

Objective To explore the expression of thioredoxin reductase (TrxR) mRNA and protein,as well as the relationship between TrxR and myeloid leukemia.Methods Bone marrow samples from 20 acute myeloid leukemia (AML),20 chronic myeloid leukemia (CML),and 20 healthy persons as normal control group were collected.The human non small cell lung cancer A549 cells were selected as the positive control group.The expression of TrxR mRNA was detected by real-time quantitative polymerase chain reaction.The expression of TrxR protein level was detected by Western blot.Results The relative quantitation expressions of TrxR mRNA were 6.751 (13.459),4.321 (11.389),18.477 (2.089),1.045 (0.467) in AML,CML,positive control and normal control group,were 17.814±3.979 and 4.860±1.550 in the primary diagnosed/relapse group and complete remission (CR) group of AML patients,and were 19.552 (5.758) and 3.459 (2.047) in the primary diagnosed and treatment group of CML patients.The expression levels of TrxR mRNA in AML,CML and positive control group were significantly higher than that in normal control group (H =43.978,P ＜ 0.001).For AML patients,the expression levels of TrxR mRNA in the primary diagnosed/relapse group,CR,positive control group were significantly higher than that in normal control group (F=246.793,P ＜ 0.001),and the difference between the primary diagnosed/relapse group and positive control group was no significant (P ＞ 0.05).The expression of TrxR mRNA were higher in the primary diagnosed/relapse group than that in CR group,and that in CR group than that in normal control group (both P ＜ 0.05).For CML patients,the expression levels of TrxR mRNA in the primary diagnosed,treatment,positive control group were significantly higher than that in normal controls (H =38.222,P ＜ 0.001),the difference between that in the primary diagnosed and that in positive control group was no significant difference (P ＞ 0.05),but the expression of TrxR were significantly higher in the primary diagnosed group than that in treatment group,and that in treatment group than that in normal controls (both P ＜ 0.05).The expression of TrxR mRNA in the primary diagnosed/relapse group of AML and the primary diagnosed group of CML showed no significant difference (P ＞ 0.05).The positive levels of TrxR protein were 0.679,0.606,0.877 and 0.095 in AML,CML,positive control and normal control group.TrxR protein was highly expressed in myeloid leukemia patients.Conclusion The expression levels of TrxR mRNA and protein are increased in myeloid leukemia.The low expression of TrxR in normal hematopoietic stem cells is highly expressed in myeloid leukemia,and is downgraded after treatment,which may be used as one of the parameters to diagnosis,effect and prognosis of myeloid leukemia.

Capillary electrophoresis with many advantages plays an important role in pharmaceutical analysis and drug screening. This review gives an overview on the recent advances in the developments and applications of capillary electrophoresis in the field of enzyme inhibitor screening. The period covers 2013 to 2017. Both the pre-capillary enzyme assays and in-capillary enzyme assays which include electrophoretically mediated microanalysis (EMMA) and immobilized enzyme microreactor (IMER) are summarized in this article.

Objective To analyze the characteristics of diagnosis and therapeutic effect of malignant pleural effusion with lung cancer,and explore the prognostic factors and effective diagnosis and treatment plans.Methods A retrospective analysis in-cludes 728 cases of definite pleural effusion with lung cancer from October 2009 to December 2013 in our hospital,which was fol-lowed-up to September 30,2014,and 438 cases were available analyzed.The main outcome measures were incidence,efficiency,pro-gression-free survival and overall survival.Results The overall median progression-free survival and the median survival of malig-nant pleural effusions with lung cancer was respectively 4 months and 8 months.males,small cell lung cancer,massive pleural effu-sion,and right pleural effusion may be the independent factors of local unmanageable malignant pleural effusion by multivariable Lo-gistic regression.The overall survival of pleural effusion with locally control(less than four weeks )was better than those with un-manageable(mOS:9 month vs.5 month,P<0.001).It was no significant difference of the control rate for overall survival prognosis between pleural effusion with two weeks and those with four weeks.histological type,partial remission time,the number of cycles of chemotherapy and TKI therapy may be the independent development risk by progression with Multivariate Cox regression analysis. the progress risk of patients with squamous cell carcinoma and other histological types were lower than that of adeno carcinoma (P=0.007).The progress risk of development of patients with 4 weeks of pleural effusion was significantly lower than that of the non remission (P=0.004),the progress risk of complete chemotherapy cycle number > 2 cycles and takingTKI treatment were significantly reduced (P<0.001;P=0.026).Gender,histological type,pericardial effusion,partial remission time,cycles of chemo-therapy and TKI were the independent prognostic factors for overall survival.The overall survival prognosis of patients with Fe-male,squamous cell carcinoma,no pericardial effusion,and over three cycles of chemotherapy,TKI therapy,and local controlled in 4 weeks was better.Conclusion Male,massive pleural effusion and right pleural effusion are independent predictive factors of local unmanageable malignant pleural effusion.The overall survival of pleural effusion with locally control was better than the patients with refractory control.Histological type,controllable relief time of pleural effusion,cycles of chemotherapy and TKI therapy were the independent predictive factors of progression and overall survival.

Objective To compare the clinical manifestations,renal histological lesions,and the levels of urinary protein markers between the children with IgA nephropathy (IgAN) and those with IgM nephropathy (IgMN), and to determine whether urinary protein markers could predict the severity of renal histological lesions in children with IgAN and IgMN.Methods Seventy-four children with renal biopsy-proven IgAN and IgMN from January 2002 to October 2014 were enrolled in the study.The levels of IgG, albumin (Alb), transferrin (TRF), α1-microglobulin (α1-MG) ,β2-microglobulin (β2-MG) and N-acetyl-β-glucosaminidase (NAG) in morning urine samples before biopsy were measured.The semi-quantitative scores of mesangial hypercellularity (MC), glomerulosclerosis (GS), and tubule-interstitial damage (TID) were used to assess renal histological lesions.Multivariate Logistic regression analysis was used to determine whether urinary protein levels were independently associated with renal histological lesions.The area under the receiver-operating-characteristic curve (AUC) was calculated to assess the predictive ability of urinary protein markers.Results Seventy-four children (44 cases with IgAN,30 cases with IgMN) were included.The urinary levels of α1-MG and Alb were significantly higher in children with IgAN as compared to those with IgMN.The differences, however, did not remain significant after adjustment for age.The urine protein, as an independent factor associated with severe MC(＞ 5 mesangial cells per mesangial area) was TRF(B =0.010), and severe GS (≥ 10％ glomeruli showing segmental adhesion or sclerosis) was significantly correlated with Alb(B =0.001) ,and severe TID (focal or diffuse tubular and interstitial lesions) was significantly correlated with NAG(B =0.038).Urinary β2-MG was not significantly associated with severe MC, GS and TID.Urinary TRF, Alb and NAG achieved the best AUC of 0.85 (P ＜ 0.001) ,0.78 (P =0.002), and 0.78 (P =0.003), respectively, for predicting severe MC, GS, and TID.Conclusions Urinary proteins are useful to predict the severity of renal histological lesions in children with IgAN and IgMN.Urinary TRF, Alb and NAG have better predictive value.

Objective To determine if urinary cystatin C (uCys C) level can predict mortality in critically ill neonates.Methods This prospective study included neonates admitted to the intensive care unit within the first 6 hours of life from May.2011 to Oct.2012.Neonates were assigned into survivor and non-survivor groups based on whether they died during the first week of life.The uCys C level was measured on the day of admission.The score for neonatal acute physiology (SNAP) was calculated based on 28 items collected during the first 24 hours of admission.Multivariate Logistic regression analysis was used to determine whether uCys C level was a predictor of mortality.A receiver operating characteristic (ROC) curve was constructed and the area under the ROC curve (AUC) was calculated to assess the predictive strength.Results Of the total 155 neonates,12 cases (7.1％) died during the first week of life.When compared to survivors,the gestational age (t =2.810,P =0.006) and birth weight (t =3.245,P =0.001) in non-survivors were significantly lower; but the uCys C level (z =-3.426,P =0.001),the SNAP score (z =-3.308,P =0.001),and the use of mechanical ventilation (x2 =23.877,P =0.000) were significantly higher.Logistic regression analysis revealed that uCys C remained significantly associated with mortality after adjusting for gestation age,birth weight,or the SNAP score (P =0.024).uCys C achieved AUC of 0.81 (95％ CI:0.71-0.92,P =0.001).When combined with SNAP and mechanical ventilation,the predictive performance of uCys C improved AUC 0.93 (95 ％ CI:0.86-1.00,P =0.000).Conclusion uCys C is significantly associated with adverse outcome of death and independently predictive of mortality in critically ill neonates.

Objective To investigate the expression of nuclear factor erythroid-2 related factor 2 (Nrf2) gene in chronic myeloid leukemia (CML) and explore its relationship with clinical characteristics and thioredoxin reductase (TrxR).Methods The expressions of Nrf2 and TrxR genes in bone marrow cells and K562 cells were analyzed in 30 bone marrow preparations of CML patients in different phases,including 20 in chronic phase,3 in accelerated phase,7 in blastic phase by real-time quantitative polymerase chain reaction.Ten health subjects were served as normal controls.Results The relative quantitation expression of Nrf2 and TrxR mRNA were 5.601±1.069 and 9.017±2.398 in chronic phase,1.698±0.349 and 5.590±1.015 in accelerated phase,1.252±0.807 and 5.050±1.469 in blastic phase,1.377± 0.344 and 1.867±0.977 in normal controls.The expressions of both Nrf2 and TrxR mRNA in CML had significant differences from those of the normal controls (x2 =14.680,P =0.002,x2 =8.271,P =0.041).The expression of Nrf2 mRNA in accelerated phase,blastic phase group showed no significant difference (Z =0.011,P =0.496),but lower than that in chronic phase group (Z =2.155,P =0.016,Z =2.534,P =0.006).The difference between the first visit and post-treated group was significant (Z =2.015,P =0.022).The expression in K562 cells and normal controls had significant difference (Z =1.898,P =0.029).In CML patients,the expression of Nrf2 was positively correlated with that of TrxR (r =0.738,P =0.037).Conclusion The expression of Nrf2 gene is higher in the first visit group of CML than that in the other groups,and is decreased after therapy,which may be the molecular marker predicting the progress of CML.Nrf2 mRNA expression level is correlated with TrxR.