Objective To investigate the variation of immune index in patients with systemic lupus erythematosus (SLE) treated with autologous purified CD+34 cells transplantation and to clarify the relationship with pathogenesis and prognosis. Methods Flow cytometry (FCM) and enzyme linked immunosorbent assay(ELISA) were used to test lymphocyte subsets, C3, C4, CH50, autoantibodies and immunoglobulin for 18 cases of SLE before and after transplantation. Results The results showed that the ratio of all the T cell subsets reduced obviously in early postgraft and recovered gradually in 1 to 3 months after transplantation except CD45 RO+CD+4 cells. The levels of serum C3, C4, CH50 increased significantly after transplantation. No case relapsed within one year after transplantation, but 2 patients relapsed one year after transplantation. The levels of the indexes in the patients with relapse were significantly lower than those in the patients with persistent remission, including C4 in the entire course, CH50 in the 3rd and 12th month after transplantation and CD45 RA+ CD+8 cells in the 6th month after transplantation. However, the ratio of CD45 RO+ CD+4 cells in the first month after transplantation in the patients with relapse was higher than that in the patients with persistent remission. Conclusion Autologous purified CD+34 cells transplantation is effective for treating SLE. Survey of immune indexes before and after transplantation is important to investigate the pathogenesis of SLE. Moreover, these immune indexes can be used to predict therapeutic efficacy of SLE.

OBJECTIVEIsobaric tags for relative and absolute quantitation (iTRAQ) combined with mass spectrometry were used to screen differentially expressed plasma proteins in cold stress rats.

METHODSThirty health SPF Wistar rats were randomly divided into cold stress group A and control group B, then A and B were randomly divided into 3 groups (n = 5): A1, A2, A3 and B1, B2, B3. The temperature of room raising was (24.0 +/- 0.1) degrees C, and the cold stress temperature was (4.0 +/- 0.1) degrees C. The rats were treated with different temperatures until 12 h. The abdominal aortic blood was collected with heparin anticoagulation suction tube. Then, the plasma was separated for protein extraction, quantitative, enzymolysis, iTHAQ labeling, scx fractionation and mass spectrometry analysis.

RESULTSTotally, 1085 proteins were identified in the test, 39 differentially expressed proteins were screened, including 29 up-regulated proteins and 10 down-regulated proteins. Three important differentially expressed proteins related to cold stress were screened by bioinfonnatics analysis (Minor histocompatihility protein HA-1, Has-related protein Rap-1b, Integrin beta-1).

CONCLUSIONIn the experiment, the differentially expressed plasma proteins were successfully screened in cold stress rats. iTRAQ technology provided a good platform to screen protein diaguostic markers on cold stress rats, and laid a good foundation for further. study on animal cold stress mechanism.

Animals ; Blood Proteins ; chemistry ; Cold Temperature ; Mass Spectrometry ; Rats ; Rats, Wistar ; Stress, Physiological

Objective: To analyze the relationship between thrombin activatable fibrinolysis inhibitor ( TAFI ) and coronary heart disease ( CHD ), and to provide evidence for the prevention of CHD. Methods: The patients with CHD in Fushun Central Hospital in Liaoning Province were selected as the case group, the patients without CHD in the same hospital and period were selected as the control group. The demographic information and clinical examination results ( serum TAFI, lipid, glucose, etc. ) were collected to analyze the association between TAFI and CHD by logistic regression models.The multivariate logistic regression analysis was used to explore the relationship between TAFI and CHD. Results: There were 222 cases, including 100 cases of stable angina, 44 cases of unstable angina and 78 cases of acute myocardial infarction, and 222 controls. The median ages of cases and controls were 62 and 57 years old. The results of multivariate logistic regression analysis showed that serum TAFI>22.88 μg/mL ( P75 of controls ) was associated with the risk of CHD ( OR=1.619, 95%CI: 1.011-2.593 ), unstable angina ( OR=2.917, 95%CI: 1.433-5.939 ) and acute myocardial infarction ( OR=2.626, 95%CI: 1.007-6.847 ). Conclusion The high level of TAFI is related to CHD, unstable angina and acute myocardial infarction.

OBJECTIVESTo investigate the incidence of intra- and extrauterine growth retardation (EUGR) and growth restriction in premature infants, and to illustrate the growth pattern of them in postnatal and infantile period.

METHODSAll premature infants were admitted to our neonatal intensive care unit (NICU) during the recent 7 years. The criteria for enrollment were (1) gestational age < 37 weeks; (2) single fetus; (3) admitted within the first 24 hours of life; (4) hospitalization period ≥ 14 days; (5) clinical follow-up persisted till ≥ 3 months of corrected gestational age. Intrauterine growth restriction (IUGR), EUGR and growth restriction were defined as having a measured growth value (weight) that was ≤ 10(th) percentile of Chinese infants' growth curve in corrected age on admission, discharge and follow-up period. Results were analyzed by using SPSS 12.0 statistical software package by chi-square test, rank-sum test, and t test.

RESULTSTwo hundred and thirty nine infants were involved, 134 were boys and 105 girls. The incidence of IUGR and EUGR assessed by weight was 25.5% and 40.6%, respectively. The lower the birth weight was, the higher the incidence of IUGR and EUGR was. The percentile of body weight in the growth curve at discharge was lower than that at birth (Z = -7.784, P = 0.000). The incidence of growth restriction assessed by weight was 20.5%, 15.0%, 8.8%, 17.0%, 10.4%, 10.1%, 11.9%, 7.0% at corrected gestational age of 38 - 40 weeks, corrected age of 28 d, 61 d, 91 d, 122 d, 152 d, 183 d, and 274 d, respectively. The incidences of growth restriction were stable when the corrected age was older than 91 days. The incidence of growth restriction in female premature infants at 183 days' corrected age was higher than that in male children (χ(2) = 6.181, P = 0.017), the incidence was 19.3% and 3.8% respectively. During the follow-up period, most of the average body weight of premature infants whose gestational age was < 32 weeks or birth weight ≤ 1500 g were lower than the 50(th) percentile of the growth curve except the average body weight of boys whose gestational age < 32 weeks at corrected age of 2 and 4 months.

CONCLUSIONSPremature and/or low birth weight infants are at high risk of growth restriction, especially very low birth weight infants. The incidence of growth restriction decreased with growth. Long-term prognosis requires further investigation.

Body Weight ; Female ; Fetal Growth Retardation ; Follow-Up Studies ; Humans ; Infant, Newborn ; Infant, Premature ; growth & development ; Male ; Weight Gain

OBJECTIVETo study the relationship of protein intake and energy supply with the physical growth in very/extremely low birth weight infant at their early life.

METHODRetrospective survey was performed in Neonatal Intensive Care Unit (NICU) in Peking University First Hospital. Inclusion criteria were preterm infant, birth weight < 1500 g, hospitalization for longer than 2 weeks, discharge with body weight greater than 1800 g. The infants were divided into two groups according to gestational age (GA). GA < 32 weeks and ≥ 32 weeks. Physical growth and its relation with the protein intake and energy supply were analyzed. The predictive value of serum blood urea nitrogen (BUN) on protein intake was studied.

RESULTNinety-three very/extremely low birth weight infants were involved, 69 in GA < 32 weeks group and 24 in GA ≥ 32 weeks group.Compared with GA ≥ 32 group, GA < 32 weeks preterm infants had more weight loss, (9.2 ± 4.4)% vs. (5.0 ± 3.1)%, P = 0.000; slower birth weight recovery (10.6 ± 3.8) d vs. (7.1 ± 2.6) d, P = 0.000; poorer weight gain at 1, 4, 5 weeks of life, (-4.5 ± 9.3) g/ (kg·d) vs. (3.4 ± 6.9) g/ (kg·d), P = 0.000 , (13.5 ± 7.3) g/ (kg·d) vs. (19.2 ± 4.9) g/ (kg·d), P = 0.001, (14.6 ± 5.6) g/ (kg·d) vs. (18.2 ± 4.5) g/ (kg·d), P = 0.031; less energy supply at 1 to 5 weeks (P value was 0.000,0.000,0.025,0.001,0.008 respectively) and less protein intake at 1, 4, 5 weeks of life (P value was 0.009,0.006,0.032). Extrauterine growth retardation (EUGR) was still predominant in our subjects, 47.8% in GA < 32 weeks group, and 95.8% in GA ≥ 32 weeks group, P = 0.000. The incidence increased greater in GA < 32 weeks infants, 43.5% vs. 20.8%, P = 0.000.The duration of weight loss and mechanical ventilation correlated negatively with weight gain rate, respectively β = -0.591, P = 0.000 and β = -0.281, P = 0.005; the average energy supply and time taken to reach full enteral feeding were factors improving weight gain, respectively β = 0.202, P = 0.021 and β = 0.354, P = 0.000. After birth, serum BUN declined gradually. Positive relation showed between average protein intake at 3(rd) week and BUN level at the end of 3 weeks, r = 0.420, P = 0.000. Serum BUN 1.44, 1.49 mmol/L at the end of 3(rd) and 4(th) week were cut-off predictors for protein intake less than 3 g/(kg·d) at related period, sensitivity and specificity were 65.3%, 83.3% and 60%, 80% respectively.

CONCLUSIONNo enough protein intake and energy supply, poor weight gain are critical problems in the management of very/extremely low birth weight infants. Prevention from NEC, appropriate parenteral/enteral nutrition transforming will benefit their physical growth. Low serum BUN after 3 weeks of life is a valuable predictor of low protein intake.

Blood Urea Nitrogen ; Dietary Proteins ; administration & dosage ; Energy Intake ; Enteral Nutrition ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Infant, Low Birth Weight ; growth & development ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases ; epidemiology ; etiology ; Infant, Very Low Birth Weight ; growth & development ; Intensive Care Units, Neonatal ; Nutritional Status ; Parenteral Nutrition ; Retrospective Studies ; Weight Gain

INTRODUCTIONThis study aimed to determine the early growth patterns of preterm infants who required prolonged hospitalisation in terms of body weight Z-score, and to explore the influencing factors and predictors of their growth.

METHODSThe criteria of enrolment included preterm birth, singleton pregnancy, hospitalisation within the first 24 hours of life, hospital stay ≥ 28 days and clinical follow-up beyond 91 days of corrected age. Body weight Z-scores and the incidence of underweight infants were reviewed periodically, and the influencing factors and possible predictors of growth analysed.

RESULTSBody weight Z-scores of all infants of gestational age (GA) groups kept decreasing, with a trough seen at 36 weeks corrected gestational age (CGA). At corrected full-term, body weight Z-scores for all birth weight groups achieved birth level and were higher than that at 36 weeks CGA. Body weight Z-scores at 61 days corrected age was (-0.300 × GA [weeks] + 0.210 × birth weight [g] + 0.682 × body weight Z-score) at 40 weeks CGA. The cut-off values for body weight Z-score at birth (cut-off, -1.79; sensitivity, 100%; specificity, 91.3%) and 61 days corrected age (cut-off, -1.95; sensitivity, 100%; specificity, 97.1%) were selected to predict the risk of being underweight at 183 days corrected age.

CONCLUSIONEarly growth restriction is a practical problem in preterm infants with prolonged hospitalisation. Body weight Z-scores at 40 weeks CGA and 61 days corrected age can be used to predict body weight gain prior to 183 days corrected age in these infants.

Female ; Follow-Up Studies ; Gestational Age ; Growth Disorders ; epidemiology ; etiology ; Humans ; Incidence ; Infant, Newborn ; Infant, Premature ; growth & development ; Infant, Premature, Diseases ; epidemiology ; etiology ; Length of Stay ; trends ; Male ; Pregnancy ; Retrospective Studies ; Singapore ; epidemiology

Abstract?AIM: To investigate mechanism of bradykinin ( BK) on inflammations of retinal pigment epithelium ( RPE) cells.?METHODS: ARPE -19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy.The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.?RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology.Kinin B1 receptors ( B1R ) and B2 receptors ( B2R ) could be detected in ARPE-19 cells.Compared with control group, Ca2+concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+concentrations.Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group (P<0.001).?CONCLUSION:BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

Aim To investigate the effects of TRPV4-Nox2 complex on ROS production and aortic vasodilatory function in mice fed with high-fat diet.Methods Male C57 BL/6J mice and TRPV4 KO mice were randomly divided into seven groups, with 10 mice in each group: normal diet group(ND), high-fat diet group(HFD), TRPV4 KO mice fed with high-fat diet group(TRPV4 KO-HFD), HFD+AAV-Flt1-Vector/Nox2 ▵3 group, TRPV4 KO-HFD+AAV-Flt1 -Vector/Nox2 ▵3 group.Body weight and blood pressure were recorded.14 weeks later primary aortic endothelial cells were isolated for CM-H2DCFDA staining and immuno-FRET assay, and aortic rings were isolated for vascular tone assay.Results ① Obesity significantly increased ROS production, triggered vasodilatory dysfunction and increased the strength of physical coupling between TRPV4-Nox2 complex(P<0.05); ② Decreasing the physical association of TRPV4-Nox2 complex could help reduce obesity-induced increased ROS production and vasodilatory dysfunction(P<0.05); ③ Entrectinib had no effect on the expression and function of TRPV4 and Nox2, but only decreased the physical association of the TRPV4-Nox2, which in turn improved obesity-induced oxidative stress and restored vasodilatory function.Conclusions Reducing the physical association of TRPV4 and Nox2 through Entrectinib can help reduce obesity-induced increase in ROS production and improve vasodilatory function of obese mice.

Tn order to set up a mouse model of myelofibrosis (MF) induced with high dose recombinant human erythropoietin (rhEPO). 60 mice were collected and divided into EPO and control groups, the former was injected with rhEPO and the latter with normal saline intraperitoneally. 5 mice from each group were executed on day 6, 30, 60, 90, 120 and 150 respectively. Their WBC count, Hb level, MCV, RDW and platelet amount were measured by automatic blood cell analyzer; CD34(+) cell ratio in bone marrow were analyzed by flow cytometry; liver and spleen coefficients were measured; pathological changes of liver, spleen, femur were observed by HE staining and reticular fibers staining; cortex thickness, femoral canal diameter and lumbar spine density were determined by computerized tomography (CT). The results indicated that as compared with normal control group in EPO induced group, WBC count was increased slightly in whole period, but without statistic significance (p > 0.05), Hb level and RDW increased at day 6 and 30 significantly (p < 0.05), MCV increased at day 6 significantly (p < 0.05), but platelet amount decreased significantly at all time points (p < 0.05). Most mice in EPO-induced group had hepatomegalia and their liver and spleen coefficient increased significantly at day 60 (p < 0.05), while most mice had splenomegaly and its coefficient was increased significantly at all time-points (p < 0.05). CD43(+) cell ratio of EPO group increased significantly in whole period (p < 0.05). CT scanning displayed femoral cortical thickening, medulla canal narrowing and lumbar spine density increasing at day 150, meanwhile, HE staining and reticular fiber staining showed the fatty degeneration or vacuolization in liver, splenomegaly with megakaryocytic proliferation, femur bone marrow fibrosis and osteosclerosis. It is concluded that the mouse induced by high dose of rhEPO displays the myelofibrosis associated with splenic extramedullary hemopoiesis, and this study is useful to establish a practical MF model, and to explore its pathological mechanism.
Animals ; Disease Models, Animal ; Erythropoietin ; adverse effects ; Female ; Humans ; Mice ; Mice, Inbred Strains ; Primary Myelofibrosis ; chemically induced ; Recombinant Proteins ; adverse effects