BACKGROUND: Decreasing of absorption of zinc from small intestine can induce dermatitis, alopecia, growth and developmental disorder and so on. It is not very clear that how to keep homeostasis when there is low zinc concentration. The discoveries of zinc transporters and relative researches provide new study direction.OBJECTIVE: To study the effect of different zinc concentration on the expressions of divalent metal transporter 1 (DMT1) and human zinc-regulated transporter (ZRT), iron-regulated transporter (IRT)-like protein (ZIP) 4mRNA, and analyze the potential pathway of absorption of zinc from small intestine in low zinc concentration.DESIGN: Blank-controlled experiment.SETTING: Department of Military Hygiene, Navy Faculty, Second Military Medical University of Chinese PLA.MATERIALS: The experiment was accomplished in Department of Military Hygiene, Navy Faculty of Second Military Medical University of Chinese PLA between October 2004 and May 2005. Materials were human colon adenocarcinoma cells Caco2 that were purchased from Shanghai Institute of Cell of Chinese Academy of Sciences.Time-dependent effect: The expression of DMT1 and ZIP4 mRNA was detected with reverse transcription-polymerase chain reaction (RT-PCR) at 0,dependent effect: The expression of DMT1 and ZIP4 mRNA were measured after the 0, 2.5, 5, 7.5 and 10 μmol/L TPEN exposure, respectively, by RT-PCR.MAIN OUTCOME MEASURES: Time-and dose-dependent effect of zinc on the expression of DMT1 and ZIP4 mRNA in Caco2 cells.RESULTS: ①Time-dependent effect: Compared with 0 hour, the expression of DMT1 mRNA obviously increased at 6, 8 and 10 hours (P ＜ 0.05 ), but there was no significant change at 2 and 4 hours. The expression of ZIP4 mRNA markedly increased in all timing, and ZIP4 mRNA level at the 6th hour reached the peak with the prolongation of low zinc duration,which was 2.1 times ofthat at 0 hour. ②Dose-dependent effect: The expression of DMT1 mRNA distinctly increased at the concentration of 7.5 and 10 μmol/L TPEN exposure as compared with that of the control group (P ＜ 0.05), but there was no significant change at 2.5 and 5 μmol/L. The ZIP4 mRNA expression increased with the increasing of the concentration of TPEN, the expression of ZIP4 mRNA was 2.7 times of that at 0 μmol/L. CONCLUSION: Zinc can regulate the expression of DMT1 and ZIP4 mRNA in Caco2 cells, and there is time-and dose-dependent effect. But the mRNA expression of ZIP4 is more sensitive and prompt than DMT1. Cells can upregulate the expression of DMT1 and ZIP4 mRNA to keep the homeostasis in low zinc condition.

Objective To investigate the effect and mechanism of FRNK on the proliferation of Hepatic Stellate Cells in vitro.Methods FRNK plasmid mediated by cationic liposome was transfected into HSCs.The proliferation of HSCs was evaluated by modified MTT assay.The level of FRNK,FAK,p-FAK(Tyr397),ERK1 and p-ERK in HSCs were assayed by Western blot and RT-PCR.Results Compared with nFRNK plasmid group,FRNK inhibited the proliferation of HSCs and the inhibition rates at 12,24 and 48 h were 20.07%,26.16% and 29.77%(P

Objective To observed the expression of matrix metalloproteinase-9 in the early onset of severe acute pancreatitis associated with acute lung injury in rats and investigate its effection in lung injury.Methods Thirty-two healthy adult male SD rats were randomly divided into two groups:Control group (n =8),Severe acute pancreatitis group(n =24).Severe acute pancreatitis model was induced by retrograde inject the 4％ sodium taurocholate sodium taurocholate into the biliopancreatic duct of rats.The severe acute pancreatitis group was detected the rate of lung water content、arterial blood gas.myeloperoxidase,matrix metalloproteinase-9,histopathology of the pancreas and lung injury score under the light microscope at 3 hours,6 hours and 12 hours.The matrix metalloproteinase-9 expression was detected by immunohistochemical and the results of immunohistochemical were analysed by the Image-Pro Plus image analysis system.Control group was detected the relevant indicators at 12 hours.Results Successfully modeling,the expression of matrix metalloproteinase-9 gradually increased beginning at 3 hours,at twelve hours up to the highest value(P ＜ 0.05).The degree of lung injury,lung water content,myeloperoxidase activity,PaCO2 gradually increased(P ＜ 0.05),PaO2 decreased significantly P ＜ 0.05).Conclusions The high expression of matrix metalloproteinase-9 is important to the pathogenesis of severe acute pancreatitis associated with acute lung injury.

Objective:To observe the growth and development of the weaned mice fed with different levels of dietary zinc and to explore the expression of zinc transporter 3(ZnT3) mRNA induced by different dietary zinc intakes. Methods: Twenty male weaned mice (postnatal day 21) were divided into 4 groups: zinc deficient (ZD), zinc adequate(ZA), zinc supplemental (ZS) and pair fed(PF). Mice were fed with different levels of dietary zinc for 3 weeks (from postnatal day 21 to postnatal day 42) ;the zinc contents of ZD, ZA, ZS and PF group were 1 mg/kg, 30 mg/kg, 180 mg/kg and 180 mg/kg respectively. From postnatal day 21 to postnatal day 42, the diet intakes and weight of the mice were measured everyday. On postnatal day 42, the mice were sacrificed and tissues were immediately isolated and frozen lor RNA extraction. The serum zinc concentrations were measured by AAS and the expression of ZnT3 mRNA was determined by semiquantitative RT-PCR. Results: The dietary intakes and weight of ZD mice were much lower than that of other groups(P3

AIM:To evaluate the inhibitory effect of FRNK on the phosphorylation of FAK and apoptosis in hepatic stellate cells (HSCs). METHODS: After stimulated with fibronectin, HSCs was transfected with FRNK plasmid by cationic liposome method. The apoptosis of FRNK-induced HSCs was examined by Annexin-V/propidium iodide double-labeled flow cytometry (FCM), gel electrophoresis and transmission electron microscope. The protein levels of FRNK, FAK and p-FAK (Tyr397) in HSCs were assayed by Western blotting, and RT-PCR was used to detect the expression of mRNA. RESULTS: The expression of FRNK was enhanced and the phosphorylation of FAK was inhibited after FRNK was transiently transfected into HSCs in vitro. The apoptotic rate in HSCs exposed to FRNK plasmid for 48 h was higher than that in the non-FRNK plasmid group [(25.37?1.92) % vs (9.28?1.05) %, P

Objective To investigate the effects of recombinant adeno-associated virus-mediated myostatin propeptide (MPRO) on uptake and oxidation of glucose,and glycogen synthesis in C2C12 myotubes,as well as the associated molecular mechanism.Methods Mature C2C12 myotubes were assigned to the following 6 groups:control,insulin,green fluorescent protein (GFP),insulin + GFP,MPRO,and insulin + MPRO groups.Glucose uptake,glucose oxidation,and glycogen synthesis were detected by counting radioactivity of 14CO2 or 14C labeled glycogen derived from 2-deoxy-[1-14 C] glucose.The activity of insulin signal pathway was evaluated by Western blot.Results Compared with control group,glucose uptake and glycogen synthesis were significantly increased in insulin and insulin+GFP groups,and further increased in insulin+MPRO group as compared with insulin alone(all P＜ O.05).However,MPRO and insulin had no effect on glucose oxidation.The phosphorylations of insulin receptor (IR) β,insulin receptor substrate 1 (IRS-1),protein kinase B (Akt),glycogen synthase kinase-3 β (GSK-3β),and the expressions of phosphatidylinositol 3-kinase (PI3K) and glucose transporter 4 (Glut4) in membrane were significantly increased in insulin and insulin+GFP groups compared with control group(all P＜0.05),and were further increased after MPRO transfection (all P ＜ 0.05).Conclusion MPRO may increase insulin-stimulated glucose uptake and glycogen synthesis in C2C12 cells by activating the IRS/PI3K/Akt signal pathway.

Objective To investigate the effects of electroacupuncture (EA) on the implementation of blind nasojejunal (NJ) tube placement and enteral nutrition (EN) in neurosurgical severe coma patients in intensive care unit (ICU).Methods Seventy-nine neurosurgical severe coma patients admitted to ICU were randomly divided into conventional group (blind NJ tube placement,n =40) and EA group (NJ placement and EA,n =39).EA was performed after NJ tube placement at bilateral acupoints Zusanli (ST36) and Hegu (L14) points using EA treatment instrument.The impelling distance of NJ tube were measured and the success rate of NJ tube placement were calculated.The postoperative complications were observed.Results The difference of NJ tube impelling distances at the 24th,48th,and 72th hours after surgery in EA group were significant longer than that in conventional group (P ＜ 0.05).The success rates of NJ tube placement at the 24th and 72nd hours after surgery in EA group were significantly better than that in conventional group (P ＜ 0.05).Their EN calories qualifiedness rate in 72 hours also increased significantly compared with conventional group and the proportion of patients assisted with parenteral nutrition decreased (P ＜0.05).The postoperative complications including alimentary tract hemorrhage,vomiting,and abdominal distension decreased remarkably in EA group compared with conventional group (P ＜ 0.05).Conclusions EA stimulation at acupoints could promote the gastrointestinal peristalsis of neurosurgical severe coma patients and elevate the success rate of blind NJ tube placement,so it is beneficial for the implementation of early enteral nutrition (EEN).

AIM: To investigate the effect of FAK - related non - kinase ( FRNK) on the expression of membrane - type matrix metalloproteinase -1 ( MT1 - MMP) in hepatic stellate cells ( HSC). METHODS:FRNK were trans-fected into HSCs by cationic liposome method. The protein levels of FRNK in HSC were assayed by Western blotting. The levels of MT1 - MMP were determined by RT - PCR for mRNA and by Western blotting for protein, respectively. RESULTS: The up -regulated expression of FRNK protein was observed and it was at 48 h after transfection that the FRNK protein content was the highest ( P < 0.05 ). The expressions of MT1 - MMP mRNA and protein were also up - regulated by the transfection of FRNK, and it was at 48 h after transfection that the MT1 - MMP protein content was significantly increased. CONCLUSION: The mRNA and protein of FRNK were over - expressed in HSC transfected with the gene of FRNK. The inhibitory effect of FRNK on the collagen synthesis in HSC may be through the up - regulation of MT1 - MMP.

Objective To evaluate the correlation between ultrasonography and histopathology in acute lung injury of rats.Methods Twenty male Wistar rats were randomly divided into control group (4 rats) and experimental group (16 rats),in the experimental group acute lung injury models were step-by-step induced by oleic acid (OA) and sequentially by OA and lipopolysaccharide (LPS).After the end of each step of models,ultrasound examination was applied,and the rats were sacrificed to take the lung specimens to be observed.The ultrasonography and pathological results were analyzed,and the numbers of alveolar were counted under microscope (200 times).Results With the aggravation of acute lung injury,the sonographic findings showed unclear or disappeared pleural line,close-set of B-lines,and pulmonary consolidation with air bronchogram.For control group,the ultrasonographic score was 0.25 ± 0.50,lung injury pathological score was 0.50 ± 0.58,and the numbers of alveolar were 25 ± 3.For OA group,the ultrasonographic score was 2.86-± 1.35,lung injury pathological score was 6.28-± 0.76,and the numbers of alveolar were 10-± 2.For OA + LPS group,the ultrasonographic score was 5.83-± 2.32,lung injury pathological score was 9.83 ± 0.98,and the numbers of alveolar were 6-± 2.All 3 groups were significant different on ultrasonographic score,pathological score and alveolar counts (P ＜ 0.01).Conclusions The ultrasonography image of acute lung injury of rats were positively correlated with its histopathological alteration.The ultrasonography can effectively evaluate the degree of acute lung injury of rats.

Objective To determine the purity of sodium nitrite(NaNO2) using in stray light standard substance which was applied to check semiautomatic clinical chemistry analyzer.Methods Titration was used to determine the purity of NaNO2,the uncertainty of the purity was assessed,and a parallel test was used to validate the uncertainty.Results The purity of NaNO2 was 99.81%,and its uncertainty was 0.86%.Conclusion The uncertainty of the purity passes the validation,it shows that the purity test is credible,and its uncertainty assessment is logical.