Background The retina microglia play a eliminating effect on apoptotie cells in the neural retinal layer of normal rats during postnatal development.Milk fat globule epidermal growth factor 8(MFG.E8)can combine specifically with phosphatidylinositol serine of the surface of apoptotie cells and enhance macrophage phagoeytosis of apoptotic cells.Objective Present study was to evaluate the localization and expression of MFG-E8 and its relevant cytokines in the neural retinal layer of normal rats during postnatal development Methods Normal royal college of surgeon(RCS)rats were divided into P0,P3,P7,Pi4,P30,P45 groups according to their postnatal days,and the 30-day-old RCS rats(2 rats)served as controls.Double stain of M FG.E8 and microglial cells marker(CD11b)was performed by immunofluorescence.Expressions of MFG-E8,integrin β5,CD11b and interleukin-6(IL-6)mRNA in the neural retina were analyzed by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR).The utilization of animals complied with the Regulation for the Administration of Affair Concerning Experimental Animals by State and Science and Technology Commission.Results MFG-E8 and CD11b were positively co-expressed in retinal ganglion cell layer and external plexiform layer with the green fluorescence for FITC-labeled IgG and red fluorescence for cy3-labeled lgG respectively in normal adult rats.RT-PCR showed that the mRNA of MFG-E8,integrin 85,CD11b and IL-6 was detectable at P0 rats.The expression level of these eytokines began to rise fterward and reached peak value at P14 rats and then declined gradually,showing significant differences among different ages groups in various cytokines mRNA expression(all P<0.05).Conclusion MFG-E8 can be specifically expressed in the neural layer of retina microglia in RCS rat.

Objective To investigate the of 5-fluorouracil effects on the expression of Smad7,TGF-β receptorⅠ,Bcl-2 and Bax in keloid fibroblasts.Methods After primary culture of keloid fibroblasts,4-6 passages of cells were inoculated in 5 different concentrations of 5-fluorouracil(10,20,40,80,160μmol/L)for 24,48 and 72 hours.Proliferative ability of keloid fibroblasts was detected by MTT assay.Expression of Smad 7,TGF-βreceptorⅠ,Bcl-2 and Bax in keloid fibroblasts was measured by Western blot.Results During MTT,5-fluorouracil did not affect cell viability at 24 hour at the concentration of 10 and 20 μmol/L.Compared with the control group,no significant difference was detected(P＞0.05).At other concentrations,fibroblast death was visible in each group(P＜0.01).Western blot analysis showed that the expression of Smad7 significantly decreased and the expression of TGF-β receptor Ⅰ significantly increased in the TGF-β1 group compared with the blank control group(P＜0.0 1).5-fluorouracil could significantly enhance the expression of Smad7(P＜0.01).There was a remarkable decrease of the Bcl-2 expression and marked increase of the Bax expression in different concentrations of 5-fluorouracil compared with the control group(P＜0.05).But,5-fluorouracil did not show any effect on the synthesis of TGF-β receptor Ⅰ.Conclusion 5-fluorouracil could inhibit proliferation and induce apoptosis on human keloid fibroblasts in vitro.

Objective To investigate the expression of human bone morphogenetic protein-7(hBMP-7)in rat bone malTOW stromal cells(BMSC)with a recombinant adenovirai vector carrying the hBMP-7 gene(Ad-hBMP-7) and study the effecta of Ad-hBMP-7 transfection on BMSC difierentiation.in order to explore the possibility for hBMP-7 gene therpy.Methods The rat BMSC cultured in vitro.They were divided into 3 groups:untreated group,Ad-hBMP-7 and Ad-GFP transduced treated group.The rat BMSC were transfected by Ad-hBMP-7 and Ad-GFP. The expression of hBMP-7 was detected by RT-PCR and Western blot analysis.and the alkaline phosphatage (ALP)activity of the BMSC was observed.ResulIs In the Ad-hBMP-7 transduced treated group.hBMP-7 mRNA expression wag manifested detected by RT-PCR(470 bp),Westem blot analysis demonstrated that these cells indeed produced the hBMP-7 protein(Mr.15×103);10 days after transduction treatment,most of the BMSC were had brown black particles stained positively by ALP activity.But in Ad-GFP transduced treated group and untreated group they did not.Conclusions Ad-hBMP-7 could efficiently transfect BMSC and promote the conversion to osteoblast.The expression of hBMP-7 in rat BMSC provides a basis for hBMP-7 gene therapv.

OBJECTIVETo master the distribution of male and female soldiers' body composition with bioelectrical impedance analysis (BIA) method in order to provide data for building up the standards in comparison with those of ordinary residents.

METHODSA cluster stratified sample of 5968 graduated soldiers among different armed services colleges was measured by BIA. Five percent-95% distributions with in the upper and lower limits were established on basis of the above result. The crosswise comparison was also performed.

RESULTS(1) Five percent-95% reference ranges of people in the graduating class of armed services colleges were body fat percentage (BF%): 10.30%-20.70% (male) and 19.20%-30.10% (female), body mass index (BMI): 19.30-25.70 (male) and 18.00-23.99 (female), lean body mass percentage (LBM%): 79.27%-86.69% (male) and 69.89%-80.69% (female), muscle percentage (M%): 74.24%-83.96% (male) and 65.23%-75.27% (female), bone percentage (B%): 5.01% 5.77% (male) and 4.65%-5.51% (female). (2) Soldiers in the graduating class of armed services colleges have less BF%, more B% and M% than those of ordinary residents in the same age and the same sex.

CONCLUSION(1) Soldiers in the graduating class of armed services colleges have better body composition than that of ordinary residents in the same age and the same sex; (2) Standard of body composition for soldiers in the graduating class of armed services colleges should be different from that of ordinary residents, a new standard should be built up.

Body Composition ; Female ; Humans ; Male ; Military Personnel ; Young Adult

Objective To investigate the effects of elasticity resistance (Ers) in respiratory system on oxygenation in patients with acute lung injury (ALI) after recruitment maneuvers (RM). Method Meta-analysis of data about the effects of recruitment maneuvers on oxygenation in ALI patients with different elasticity resistances in respiratory system carried out with pooling of study-oriented data stored in Pubmed, Embase, Web of Science databases from January 1999 to June 2010. Results A total of 281 articles were taken, and 20 of them included a sample size of 395 ALI patients. In patients treated with RM in different degrees of respiratory system elasticity resistance ( ≥33.3 cmH2O/L and ＜33.3 cmH2O/L), the effect of RM was better in patients with the high respiratory system elasticity resistance than that with the low one [(51.97 + 8.89) mmHg vs. (35.13 ± 10.33 ) mmHg], P ＜ 0. 01 ), but the high respiratory system elasticity resistance was potentially to lower blood pressure [(4. 33 ± 1.32 ) mmHg vs. (0.22 ± 1.03 ) mmHg],P ＜ 0.01 ). Conclusions This study suggests RM could improve oxygenation of ALI patients with high respiratory system elasticity resistance, and caution must be made to avoid hypotension during RM.

Objective To study the positive end-expiratory pressure (PEEP) adjustment after recruitment maneuver during acute respiratory distress syndrome (ARDS) especially in the presence of tonic diaphragm electrical activity (Tonic EAdi) in order to obtain optimum PEEP and in turn to get successful oxygenation.Methods Rabbit model of ARDS was nade by intratracheal instillation of hydrochloric acid.After sufficient recruitment maneuvers,the rabbits were randomly (random number) divided into two groups,namely Tonic EAdi group ( n =5 ) and maximum oxygenation group ( n =5 ).In Tonic EAdi group,the regulation of PEEP was guided by Tonic EAdi.In maximum oxygenation group,PEEP was adjusted as per maximum oxygenation.The differences in magnitude of PEEP,pulmonary mechanics,gas exchange and hemodynamics were compared between two groups.The t-test was used to compare continuous variables between the two independent samples,and the difference was statistically significant when P ＜ 0.05.Results (1) PEEP:The PEEP was (10.7 + 1.4) cmH2O (1 cm H2O=0.098 kPa) in Tonic EAdi group and (10.0 ± 2.8) cm H2O in maximum oxygenation group (P ＞ 0.05). (2) Pulmonary mechanics:After PEEP adjustment,there was no significant difference in tidal volume ( Vr),peak pressure (Ppeak) and mean pressure (Pmean) between the two groups (P ＞ 0.05 ).(3) Gas exchange:After PEEP adjustment,there was no significant difference in oxygenation index (PaO2/FiO2) and partial pressure of arterial carbon dioxide ( PaCO2 ) between the two groups (P ＞ 0.05).Conclusions Tonic EAdi could be a good indicator for regulating PEEP in ARDS.

Objective To observe the effects of recruitment maneuver (RM) and tidal volume with different amount of gas after RM ventilation on lung diastole function in rats with acute lung injury (ALI). Method ALI rat models were induced by intravenous infusion of lipopolysaccharide (LPS) in dose of 6 mg/kg. Twenty-five rats were randomly(random number) divided into control group ( n = 5), ALI group ( n = 5), low tidal volume group (LV group,VT= 6 mL/kg, n = 5), sustained inflation (SI) with low tidal volume (SI+ LV group, VT=6 mL/kg, n = 5), and SI with moderate tidal volume group (SI+ MV group, VT= 12 mL/kg, n = 5). The RM carried out by using SI with airway pressure 30 cmH-2O for 30 seconds, and the positive end-expiratory pressure (PEEP)was set at 5 cmH2O. Lung tissue was taken after mechanical ventilation for 5 hours. The mean arterial pressure (MAP) was monitored throughout the entire course of experiment. Endothelin-1 ( ET-1 ), endothelial nitricoxide synthase (eNOS), and acetylcholine-(Ach-) induced endothelium-dependent relaxation response of isolated pulmonary artery rings were investigated after mechanical ventilation for 5 hours. Results The LPS increased the ET-1 level in lung tissue, decreased the level of eNOS in lung tissue, and impaired the Ach-induced endothelium-dependent relaxation response in pulmonary vassals, without obvious influence on systemic hemodynamics. SI + LV significantly reduced LPS-induced elevation of ET-1 level, and increased the level of eNOS, and significantly lessened endothelial dysfunction and ameliorated dysfunction od endothelium-dependent relaxation in pulmonary vas sals. Conclusions RM with high tidal volume or lowtidal volume ventilation could improve the lung vascular endothelial function of rats with acute lung injury, and RM with low tidal volume ventilation could lessen more the injury of lung vascular endothelial diastole function in rats with acute lung injury.

Objective: To observe the time-effect relationship of moxibustion for primary dysmenorrhea (PD) due to stagnation and congelation of cold-damp, thus explore the optimal choice of moxibustion duration, and provide evidence for achieving satisfactory efficacy in moxibustion treatment. Methods: A total of 90 patients with PD due to stagnatin and congelation of cold-damp were divided into three groups by the random number table method, with 30 cases in each group. All the patients in the three groups were given moxibustion treatment at Guanyuan (CV 4), 20 min in group A, 40 min in group B and 60 min in group C. The changes in the pain measurement score in the three groups were observed after treatment. Results: After treatment, there were significant differences in the clinical efficacy among the three groups (P<0.05); the clinical efficacy was better in group B and group C than that in group A (P<0.05), and that in group B was better than that in group C (P<0.05). Besides, the pain measurement score changed significantly after treatment in the three groups (all P<0.05), and the between-group differences were also statistically significant (P<0.05); the pain measurement scores in group B and group C were lower than that in group A (P<0.05), and that in group B was lower than that in group C (P<0.05). Conclusion: Given the same stimulating frequency and intervention time of moxibustion, 40-minute duration demonstrates relatively better efficacy for PD due to stagnation and congelation of cold-damp.

OBJECTIVETo observe the protective effect of active fractions of Huanglian Jiedu Decoction (HJD) on primary cortical neuron injury after oxygen-glucose deprivation (OGD)/reperfusion (R) injury. Methods Using macroporous resin method, HJDFE30, HJDFE50, HJDFE75, and HJDFE95 with 30%, 50%, 75%, and 95% alcohol were respectively prepared. Then the content of active components in different HJD fractions was determined with reverse phase high-performance liquid chromatography (RP-HPLC). The OGD/R injury model was induced by sodium dithionite on primary cortical neurons in neonate rats. MTT assay was used to observe the effect of four fractions (HJDFE30, HJDFE50, HJDFE75, and HJDFE95) and seven index components of HJD on the neuron viability.

RESULTSRP-HPLC showed active component(s) contained in HJDFE30 was geniposide; baicalin, palmatine, berberine, and wogonside contained in HJDFE50; baicalin, berberine, baicalein, and wogonin contained in HJDFE75. The neuron viability was decreased after OGD for 20 min and reperfusion for 1 h, (P <0. 01), and significantly increased after administered with HJD, HJDFE30, HJDFE50, and HJDFE75 (P <0. 05, P <0. 01). Geniposide, baicalin, baicalein, palmatine, wogonside, and wogonin could increase the cortical neuron viability (P <0. 05, P <0. 01).

CONCLUSIONSHJDFE30, HJDFE50, and HJDFE75, as active fractions of HJD, had protective effect on primary cortical neuron injury after OGD/R. Furthermore, geniposide, baicalin, and baicalein were main active components of HJD.

Animals ; Berberine ; Berberine Alkaloids ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Flavanones ; Flavonoids ; Glucose ; metabolism ; Iridoids ; Models, Animal ; Neurons ; Oxygen ; metabolism ; Rats ; Reperfusion Injury ; drug therapy

This study was aimed to construct eukaryotic expression vectors carrying the small hairpin RNA (shRNA) targeting TRPC6 gene and investigate the effect of TRPC6 knockdown on puromucin aminonucleoside (PAN)-induced podocyte injury. Two DNA sequences containing the small hairpin structure targeting TRPC6 were designed, synthesized and then inserted into the green fluorescence protein (GFP)-contained plasmids (pGC) to establish the plasmids pGCsi-TRPC6A and pGCsi-TRPC6B. Plasmids expressing scrambled shRNA were used as negative control and named pGCsi-NC. These plasmids were transfected into a conditionally immortalized murine podocyte cell line by using liposome. Flow cytometry was used to examine the transfection efficiency. TRPC6 mRNA and protein expression levels were detected by RT-PCR and Western blotting. Cultured podocytes were divided into four groups: control group, PAN treatment group, PAN+TRPC6 shRNA transfected group and PAN+scrambled shRNA transfected group. The paracelluar permeability to BSA was evaluated by Millicell-PCF Inserts and cell viability was measured by the trypan blue assay. Immunofluorescent assay was used to observe the distribution of α-actinin-4 and α-tubulin. The results showed that the transfection efficiency of the shRNA expression vector was about 45%. Expression levels of TRPC6 mRNA and protein were downregulated after transfection with pGCsi-TRPC6A and pGCsi-TRPC6B. Knocking down TRPC6 gene could effectively reverse the PAN-induced increase in the paracelluar permeability to BSA. The distribution of α-actinin-4 and α-tubulin was disrupted after treatment with PAN, which was reversed by knocking down TRPC6 gene. It was concluded that knocking down TRPC6 gene could effectively prevent podocytes from the permeability increase induced by PAN, which may be related to the regulation of podocyte cytoskeleton.