Objective To investigate the normal ranges of RET-He and reticulocyte grouping parameters of healthy adults in Wenzhou. Methods A total of 1 000 samples from healthy adults were collected in Wenzhou from August 2008 to February 2010. There were 506 males and 494 females. The individuals were stratified based on age into three groups:20 to 40-year-old group (n = 341 ), ＞40 to 60-year-old group (n =316), and ＞60-year-old group (n =343). All blood samples were drawn in EDTA-K2 anticoagulated blood. Peripheral RET-He and reticulocyte grouping parameters ( IRF, LFR, MFR, HFR)were determined by Sysmex XE-2100 automated hematology analyzer. The instrument was calibrated and validated before testing All specimens were analyzed within 2 hours. RET-He, RET absolute value and RET percentage were normal distribution, the normal range was established by using ((x)-1.96s,(x)+ 1.96 s)method. IRF, LFR, MFR and HFR data were skewed distribution,the normal range was established by using (P2.5 ,P97.5 )percentile method. Results The mean and normal range of RET-He were (33.91 ± 1.77) pg and (30.44 - 37.38) pg in male, (33.97 ± 1.85) pg and (30.34 - 37.60) pg in female,respectively. There was no significant difference between the two groups ( t =-0.533, P＞0.05 ). The mean and normal range of RET absolute value were (0.051±0.023) × 1012/L and (0.006 -0.096)×1012/L in male,which were (0. 037 ±0. 026) × 1012/L and (0 -0. 088) × 1012/L in female, there was significant difference between the two groups ( t = 8. 409, P ＜ 0. 05 ) . The mean and normal range of RET percentage in male were(1.041 ±0.459)% and (0. 141 -1.941)% , and in female were (0.869±0.603)% and (0-2. 051 )%, which showed significant difference (t =5.074,P ＜0. 05). The normal ranges of LFR,MFR,HFR,IRF in male were 0. 951 (0.866 -0.988) ,0.046(0.010 -0.114) ,0. 003(0 -0.023) ,0. 050(0.012-0.134), and in female were 0.096(0.880 -0.991) ,0.039(0.008 -0.113) ,0.002(0-0.016) ,0.040(0.009-0.121), respectively. There were significant differences between the two groups (Z values were -5.044, -4.793, -3.559, -5.075, respectively,all P＜0.05). The means of RET-He in 20 to 40-year-old group, ＞40 to 60-year-old group, ＞ 60-year-old group and all age group were (33.38±1.49),(34.36 ±1.46), (34.12±2.21) and (33.94 +1.81) pg, respectively. The normal ranges were(30.46-36.30), (31.50 -37.22), (29.79 - 38.45) and (30.39 - 37.49) pg, which showed statistical difference( F =28. 072,P ＜0. 05). In addition, the normal range of RET-He in 20 to 40-year-old group was lower than that of ＞ 40 to 60-year-old group and ＞ 60-year-old group ( q values were 9. 970, 7. 791,respectively, P＜0. 05). The normal ranges of LFR, MFR, HFR, IRF in ＞ 60-year-old group were 0.961(0. 878 -0. 992) ,0. 037(0. 007 -0. 105 ) ,0. 002(0 -0. 020) and 0. 039(0. 008 -0. 117) ,while they were 0.953(0. 867 -0.990), 0.045(0.009 -0. 116) ,0.003(0 -0.019) ,0. 050(0. 010 -0. 133) in 20 to 40 -year-old group , and 0. 951 (0.855 -0.989) ,0.047 (0.010 - 0.133 ) ,0.003 ( 0-0.020), 0.049 (0.011-0.149)in ＞40 to 60-year-old group, which showed statistical difference (Z values were -3. 949, -4.236,-4. 373, -4. 973, - 2. 747, - 3. 275, - 3.901, - 4. 185, respectively, all P ＜ 0. 05). There was nodifference between male and female for RET-He, the same normal ranges could be used, but there was significant difference between different age groups. LFR, MFR, HFR and IRF were statistically different between different age groups and different sex groups. The normal ranges should be established by gender and age. Conclusion The normal ranges of RET-He and reticulocyte grouping parameters about healthy adults in Wenzhou are established, which plays an important role in clinical value of RET parameters.

To observe the effects of Niupo Zhibao Pellet, a compound traditional Chinese herbal medicine, on high-mobility group box-1 protein (HMGB1) expression in lung tissues of rats with endotoxin shock.

OBJECTIVE: To investigate the effect of Niupo Zhibao Pellet (NPZBP) on the expression of neuronal nitric oxide synthase (nNOS) in the brain of endotoxin-induced shock rats. METHODS: SD rats were randomly divided into normal control group, endotoxin-induced shock model group and NPZBP-treated group. Lipopolysaccharide (LPS) (1.5 mg/kg i.v.) and tsD-galactosamine (D-GalN) (100 mg/kg i.p.) were administered to the rats in endotoxin-induced shock model group, as well as to the rats in NPZBP-treated group after seven-day treatment, to induce the shock. The expression of nNOS in the brain of the rats in each of the 3 groups was measured by immunohistochemical methods. RESULTS: In the 3 groups, nNOS immuno-positive cells distributed widely in layer II, III, IV of the cerebral cortex, the molecular layer of hippocampus, the polymorphic layer of the dentate gyrus, the reticular formation of brain stem, and the molecular, granular and Purkinje cell layer of the cerebellar cortex. The number of immuno-positive cells in the NPZBP-treated group was slightly higher than that of the normal control group, and significantly lower than that of the model group (P<0.05) in many regions of the brain, including cerebral cortex, hippocampus, brain stem and cerebellar cortex. CONCLUSION: NPZBP can inhibit the over-expression of nNOS in wide area of the brain in endotoxin-induced shock rats.

BACKGROUND: Stem cell differentiation potential is strongly correlated with culture condition. The alteration in scaffold material surface function, three dimensional (3D) structure, and addition of growth factors can control stem cell proliferation and differentiation.OBJECTIVE: To develop 3D macroporous scaffolds with optimal porosity and porous structure to provide a microenvironment that promotes the growth of multi-potent stem cells.DESIGN: Repetitive measurement.SETTING: Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine.MATERIALS: Healthy adult SD rats were provided by the Experimental Animal Center in Guangzhou University of Chinese Medicine. Chitosan and basic fibroblast growth factor (bFGF) were purchased from Sigma Corporation (St. Louis,MO).METHODS: The experiment was performed at the Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine from March 2003 to December 2006. Using a freeze-drying method, 3D macroporous scaffolds made of different ratios of chitosan-gelatin with bFGF were fabricated that could release bFGF with controlled porosity and porous structure. Bone marrow was obtained from the femur and tibia of SD rats, and mesenchymal stem cells (MSCs) were isolated, cultured and seeded on the scaffolds with bFGF. MSCs seeded on scaffolds with no bFGF served as control. The procedure during experiment was accorded with animal ethical requirements.MAIN OUTCOME MEASURES: 3D structure and release performance of the scaffolds were observed by ELISA and scanning electron microscope; the effect of 3D macroporous scaffolds that released bFGF on MSC growth and viability were observed by HE staining, MTT, cell counting and SEM.RESULTS: There was no significant difference in pore size between scaffolds with and without bFGF (P ＞ 0.05). Scaffolds with bFGF significantly improved MSC survival rate, promoted cell adhesion, proliferation, and viability compared with scaffolds without bFGF (P ＜ 0.05).CONCLUSION: The results suggest that 3D macroporous scaffolds with bFGF release improve MSC survival on scaffolds,and lay a foundation for its application in tissue engineering.

Objective To construct luciferase reporter vector containing full-length high-mobility group box 1 ( HMGB1, GenBank NM-010439) promoter for the screening of medicine. Methods The full-length HMGB1 promoter was amplified by polymerase chain reaction ( PCR) , and then was inserted into GV238 vector to construct plasmid GV238-HMGB1-P-Luc. GV238-HMGB1-P-Luc combined with internal reference plasmid pRL was co-transfected into Hela cells ( GV238-HMGB1-P-Luc group, which served as positive control group) . Plasmid pGL3-basic combined with pRL was co-transfected into Hela cells (pGL3-basic group, which served as negative control group) . Additionally, lipopolysaccharides ( LPS, 0.2 μg/mL) was used as the activator for the positive control group (LPS group), and then sodium butyrate (SB, 10 mmol/L) was used as the inhibitor for LPS group ( SB group) . At the end of experiment hour 24, luciferase activity was detected. Results The results of digestion, amplification, sequencing and identification showed that the full length of HMGB1 promoter was 2 140 bp, and the DNA sequence was correct, without mutation. Luciferase activity in GV238-HMGB1-P-Luc group was increased as compared with that of the pGL3-basic group ( P<0.05) . Luciferase activity in the LPS group was increased ( P<0.01, compared with that of GV238-HMGB1-P-Luc group) , and then was decreased after the administration of SB ( P<0.01, compared with that of the LPS group) . Conclusion A model of luciferase reporter vector containing HMGB1 promoter has been successfully constructed. Its activity can be increased by LPS, and then is in hibited by SB. The model can be used for further screening of medicine with the activities of regulating HMGB1 promoter.