Aim To observe the expression of migration inhibitory factor (MIF) in the enteric neurons,and to explore the nervous regulation on MIF activity in experimental colitis.Methods Colitis was induced in sensitized rat and mouse by 2,4-dinitrochlorobenzene(DNCB)enema.MIF activity was measured both in the mesentery lymphocyte(by MTT)and in the enteric neurons(by immunofluorescence double staining).6-OHDA was intraperitonealy (ip) administered to mouse before DNCB treatment.Norepinephrine(NE) was added to lymphocyte culture in vitro during MIF preparation.Results The expression of MIF protein in enteric neuron was increased in DNCB-induced colitis in rat.ip 6-OHDA in colitis mouse(38～150 mg?kg-1) resulted in a further increase of MIF activity than ip vehicle in colitis mouse (P

OBJECTIVE To investigate the pathogenic causes of community-acquired pneumonia(CAP) in adult patients in Tongling.METHODS A prospective study was performed on 260 consecutive adult patients with CAP in Tongling city during last three years.Bacteria culture of sputum and serological tests in paired serum samples were detected.RESULTS Of 260 patients with etiological evaluation,128(49.2%) patients had an identifiable etiology,63(24.2%) had positive outcome from sputum cultured,atypical pathogens were detected from 75(28.8%)patients.Pathogens identified in 128 patients were:Mycoplasma pneumoniae(35.4%),Chlamydia pneumoniae(17.7%) and Streptococcus pneumoniae(13.6%).6.5% All patients had mixed infection.The resistance rate of S.pneumoniae to penicillin and erythromycin was 5 and 50%,respectively.CONCLUSIONS Atypical pathogens have important role in CAP,of which M.pneumoniae is the most common pathogen.S.pneumoniae and K.pneumoniae are the commonly encountered bacteria for CAP in Tongling.

Objective To structure the model of acute carbon monoxid poisoning(ACOP)in rats. Evaluate the effectiveness of the poisoning on the pulmonary function and the significance of carbon monoxide hemoglobin(HbCO)and oxygenation index in diagnosis of acute lung injury(ALI)/acute respiratory distress syndrome(ARDS).Method Eighty healthy adult male Wistar rats were randomized into 4 groups.According to the concentration of CO,poisoning group was randomized into three groups(each group=20),group A,group B,group C.After poisoned,arterial blood was collected rapidly for arterial blood gas analysis.According to the pathological changes,the models were divided into ALI/ARDS group and non-ALI/ARDS group.Results Compared with control group,the incident rate of ALI/ARDS in group B(25%)and group C(55%)were significantly higher(P

OBJECTIVETo explore activity laws of mitochondrial complex II in patients of deficiency-cold syndrome (DCS) and deficiency-heat syndrome (DHS) under various ambient temperatures.

METHODSSubjects were recruited by questionnaire and expert diagnosis from grade 1 - 3 undergraduates at Henan College of Traditional Chinese Medicine in November 2012, and assigned to a normal control group, the DCS group, and the DHS group, 20 in each group. Their venous blood samples were collected at two different temperature conditions. Activities of mitochondrial complex II were measured by spectrophotometry.

RESULTS(1) Comparison of mitochondrial complex It under various ambient temperatures: Compared with room temperature in the same group, activity values were all increased in the normal control group at cold temperature with significant difference (P <0.05), but there was no significant difference in the DCS group and the DHS group (P >0. 05). Compared with the normal control group, activity values of complex H were reduced in the DCS group at cold and room temperatures with significant difference (P <0.05). Compared with the DCS group, activity values of complex It were increased in the DHS group with significant difference (P <0. 05). (2) Changes of adjustment rates: Compared with room temperature, the adjustment rate all rose at cold temperature in the normal control group and the DHS group with significant difference (P <0.05), but with no significant difference found in the DCS group (P >0. 05). Compared with the normal control group at the same temperature, the adjustment rate in the DHS group and the DCS group was all reduced at cold and room temperatures with significant difference (P <0. 05). There were no significant difference in the adjustment rate between the DHS group and the DCS group (P > 0. 05).

CONCLUSIONSEnvironment temperature can affect the activity of mitochondrial complex II with different influence degrees on different syndrome types of people, but its change trend are basically identical.

Relatively uniform-sized nanoparticles made of poly (lactic-co-glycolic acid) (PLGA) were prepared by premix membrane emulsification method. After the drug loading property was completed, the dynamic tissue distribution of nanoparticles was recorded. With the average particle size and span as indexes, membrane pore size, number of passing membrane times, membrane pressure, volume ratio of oil-water phase and the concentration of poly(vinyl alcohol) (PVA) in external water phase were investigated by single factor test, the optimum preparation technology of blank PLGA nanlparticles was as following: pore size of SPG membrane was 1 μm, membrane pressure was 1. 15 MPa, the number of passing membrane time was 3, the mass fraction of PVA of 2%, volume ratio of oil-water phase of 1 : 5. Prepared nanoparticles were round with smooth surface, the mean diameter was 332.6 nm, span was 0.010, the confocal laser scanning microscope (CLSM) concluded that fluorescent substance is uniform composizion in PLGA nanoparticle, and the in vivo imaging technology in mice include that the nanoparticles show good liver and spleen targeting property.
Animals ; Drug Carriers ; chemistry ; Drug Delivery Systems ; instrumentation ; Emulsions ; chemistry ; Fluorescent Dyes ; chemistry ; Lactic Acid ; chemistry ; Mice ; Mice, Nude ; Nanoparticles ; chemistry ; Particle Size ; Polyglycolic Acid ; chemistry

AIM: To construct eukaryotic expression vector of small interfering RNA(siRNA) specific to bcl-2 and investigate the effect of recombinant plasmid on suppressing bladder cancer cell growth.METHODS: siRNA of bcl-2 gene was designed according to the principle of RNAi-based medicine, and was converted into cDNA coding expression of small hairpin RNAs(shRNA) of siRNA. The cDNA was synthesized and inserted into plasmid pGenesil-1. The recombinant eukaryotic expression vectors of pGenesil-1545 and pGenesil-1555 were controlled by the U6 promoter of RNA polymerase Ⅲ, identified by the restriction map and the sequence analysis, and transfected into T24 cells. After T24 cells were transfected for 72 h, expression of bcl-2 mRNA was assayed by RT-PCR; and MTT was used to observe the proliferation of T24 cells.RESULTS: The recombinant plasmids of pGenesil-1545 and pGenesil-1555 were identified by the restriction map and the sequence analysis. The sequences completely coincided with the designs. The expression of the bcl-2 mRNA in T24 cells transfected with recombinant plasmid decreased nearly 80%, and the growth of T24 cells was suppressed significantly.CONCLUSION: The siRNA eukaryotic expression vector against bcl-2 gene is successfully constructed. It effectively downregulates the expression of bcl-2 in T24 cells and suppresses the cell growth.

Objective To investigate the apoptosis inducing effect of peroxisome proliferator-activated receptor r(PPARr)agomst on leukemic K562 cells and its mechanisms of action.Methods K562 cells in culture medium in vitro were given different concentrations of PPARragonist rosiglitazone(RGZ)(20-80 umol/L)for O,24,48 and 72h The inhibitory rate of the cells were measured by MTT assay,cell apoptosis was detected by Annexin V/PI staining,and the expression of P53 protein as well as the activity of caspase-3 were also detected.Results RGZ(over 40umoL/L)could inhibit the growth of K562 cells and cause apopto-sis remarkably,the suppression Was both in time-and dose-dependent manner.The expression of P53 pmtein was upregulated and the activity of caspase-3 Was increased concomitantly after the cells werle treated by RGZ.Conclusion PPARr agonist RGZ(over 40 umol/L)can induce apoptosis on K562 cells signifieantly,upregu-lation the expression of P53 protein as well as increasing caspase-3 activity may be one of its most important mechamisms.

Objective To detect the differentially expressed microRNAs (miRNAs) in bladder cancer tissue and normal bladder tissue. Methods Total RNA was extracted from bladder cancer tissue and normal bladder tissue by Trizol, and then miRNAs were isolated and enriched from the total RNA. Mammalian miRNA microarrays were used to analyze the differentially expressed miRNAs between the bladder cancer tissue and normal tissue. Data analysis was performed by software of LuxS-can3. 0 and SAM version 2. 1. Choose let-7 gene which was interesting to us, and validation of mi-croarray results was carried out by northern blotting. Results Compared with normal bladder tissues, there were 71 differentially expressed miRNAs in bladder cancer tissue, of which there were 38 down-regulated ones and 33 up-regulated ones. Among these miRNAs, 26 miRNAs were the most significant with 12 up-regulated and 14 down-regulated. The expression of let-7 gene in bladder cancer was down-regulated to normal bladder tissue by northern blotting, which was in agreement with the results of the miRNA microarrays. Conclusion There are differentially expressed miRNAs between bladder cancer tissue and normal bladder tissue, and let-7 gene is probably as a tumor suppressor in bladder cancer.

BACKGROUND:It is reported that bone marrow mesenchymal stem cell(BMSC)transplantation might be a promising treatment for liver fibrosis.But the mechanism is still unclear.OBJECTIVE:To observe the hepatic stellate cells apoptosis induced by BMSC transplantation,and to study the mechanism of BMSC in treating hepatic fibrosis in vivo.METHODS:CCl_4 subcutaneous injection was performed to induce rat liver fibrosis.After 8 weeks of CCU injection,20 rats which underwent successful model establishment were randomly divided into experimental group and control group,10 in each group.The experimental group received MSC transplantation via tail vein injection,and the control group were given DMEM instead.The rats were killed and the livers were harvested at three time point,the day of MSC transplantation,3 days after transplantation,and 7 days after transplantation.The hydroxyproline content was detected by HE and Masson staining,and the expression changes of α-smooth muscle actin(α-SMA)proteins were determined using immunohistochemistry.The apoptosis of hepatic stellate cells were determined by α-SMA and TUNEL(terminal dUTP nick-end labeling)dual-staining.RESULTS AND CONCLUSION:After 8 weeks of CCU injection,the hydroxyproline content increased and histology indicated progress of liver fibrosis.At 7 days after MSC transplantation,the hydroxyproline in the liver was decreased,and the liver fibrosis was alleviated in the experimental group but aggravated in the control group.Immunohistochemistry indicated that α-SMA positive cells were increased at 8 weeks after CCU injection.At day 7 after transplantation,α-SMA positive cells in the experimental group were significantly less than control group(P ＜ 0.05).At 3 days after transplantation,the hepatic stellate cells apoptosis in the experimental group was significantly aggravated compared with control group(P ＜ 0.05).This suggested that MSC transplant was an effective treatment for liver fibrosis.MSC inducing hepatic stellate cells apoptosis may be one of the mechanisms.

Objective To introduce a improved method for the production of hemoglobin liquid by use of the hemoglobin electro‐phoresis alkaline .Methods First ,we used the pipette to absorb the settlement of red blood cells from a batch of EDTA anticoagula‐ted whole blood specimens ,then dropped them into the 0 .9% saline washed human erythrocytes .With the help of the pipette nozzle we pipet from the liquid surface to bottom repeatedly ,made the red cells suspended in the liquid evenly .The samples should be cen‐trifuged and the red blood cells fully deposited at the bottom ,then poured the supernatant after centrifugation to remove the impuri‐ties in plasma and leave the allowance of red blood cells .We add distilled water or hemolysin to lysis RBC .Hands up test tubes rack using wrist gently back and forth several times until the hemolysis was clear and transparent .Results Compared the results of the modified method and the traditional method ,then the two results compared with the results of HPLC recommended by international association of thalassemia method .Compared the results of three screening methods with that of the thalassemia gene identification method .So we could objectively evaluate the reliability of the test .Conclusion The result of the improved method are same to the electrophoretogram by SOP operation ,it will be more efficient and reduce the cost of reagent .The whole process of preparing hemo‐globin solution doesn′t contact with any chemical reagent .So there will be no pollution to the environment ,and it also reduces the harmful of toxic reagent to the human body .