Objective To investigate expression and significance of matrix metalloproteinase-2(MMP-2),MMP-9 and tissue inhibitor of metalloproteinase-1(TIMP-1) in rats with glomerular sclerosis made by doxorubicin.Methods Forty Wistar male rats(8-week-old) were randomly assigned into 2 groups:sham operated and model groups.Rats in model group were nephrectomized after anesthesia and injected with adriamycin(5 mg/kg) after 1 week.Rats in sham operated group was subjected to sham operation and injected with normal saline after 1 week through the tail vein.All rats were killed in the 12th week.Immuno-histochemistry was performed on renal tissue to detect Collagen Ⅳ(Col-Ⅳ),fibronectin(FN),MMP-2,-9 and TIMP-1.Results Immunohistochemistry staining indicated that expressions of MMP-2,-9 in model group decreased significantly compared to sham operated group(Pa

Objective To study the expression of ?-smooth muscle actin(?-SMA)in glomerulosclerosis rats and its relationship with renal function.Methods Forty healthy Wistar rats were equally divided into 2 groups including sham operated group and model control group.Rats in model groups were uninephrectomized and injected with daunorubicin(5 mg/kg)after 1 week through the tail vein.Twenty-four hours of urinary protein excretion,serum creatinine(Scr),blood urea nitrogen(BUN)were measured at the 12th week.Renal pathology was evaluated.Immunohistochemistry(SupervisionTM)was performed on renal glomeruli tissue to detect the expression of ?-SMA.Reverse transcription polymerase chain reaction(RT-PCR)was used to examine the expression levels of ?-SMA mRNA in glomeruli.SPSS 13.0 software was used to analyze the two variables.Results In model control group,the urinary protein,Scr,BUN significantly increased(Pa

Objective To establish a method for determination of trace bromate in flour and the products by ion chromatography.Methods Conductance-ion chromatography and Ionpac~R AS9-HC column were employed and 9.0 mmol/L sodium carbonate was used as the eluent,the flow rate was 1.0 ml/min.BrO_3~-in the samples soaked in the purified water was extracted by ultrasonic microwave,the water-soluble macromolecules were removed by ultra filtration cup in the extract,then the direct injection determination was conducted.Results The lowest detection limit was 0.01 mg/L,the quantitative detection limit was 0.03 mg/kg,the relative standard deviation was 0.6%-7.6%,the recovery rate was 97.8%-103.6%.Conclusion This method is simple,rapid,accurate,sensitive and is applicable to the determination of trace bromate in flour and the products.

Aim To investigate the effects of all-trans retinoic acid and benazepril on the expression of ?-smooth muscle actin in rats with glomerulosclerosis.Methods 80 Wistar male Rats were randomly assigned into the following groups: control group,model group,ATRA treatment group and benazepril treatment group,20 rats in each group.GS rats were uninephrectomized and injected with adriamycin(5mg?kg-1) after one week through the tail vein.All rats were sacrificed at the 12th week,GS was evaluated by glomerulosclerosis index(GSI) system.The expression of ?-SMA was assessed by reverse transcription-polymerase chain reaction(RT-PCR) and immunohistochemistry.Results Comparing with control group,the expression of ?-SMA mRNA and protein were decreased significantly in ATRA treatment group and benazepril treatment group(P0.05).Conclusions The postponed effects of ATRA and benazepril on GS were evident and equivalent.

The FTIR method was used to investigate the correlation of bacteria in the contaminated drug and the environmental microbes in the clean room for pharmaceutical microbial test. The similarity of bacteria in the contaminated drug and environmental microbes was compared by critical hit value method and cluster analysis method. This constructed the FTIR spectra library of clean room environmental microbe, and determined the criterion to promptly judge if the bacteria isolated from pharmaceuticals were contaminated by environment or not, hence the exactness of "one-off report" of sterile test result can be guaranteed, and can be used for the dynamic monitoring of environmental bacteria of clean room. The method is proven to be simple, accurate and rapid, and can be easily spread to the pharmaceutical microbial control.
Bacteria ; isolation & purification ; Cluster Analysis ; Drug Contamination ; Drug and Narcotic Control ; Environment, Controlled ; Environmental Microbiology ; Spectroscopy, Fourier Transform Infrared ; methods

Objective：To investigate the correct expression of dengue 2 virus 43 strain NS1 gene in transfected BHK-21 cell. Methods：The D2-43 DNA fragment coding for signal peptide plus NS1 protein was cloned between KpnⅠ site and EcoR Ⅰ site of expression plamid pcDNA3.1. The obtained recombinant vector pcDNA-NS1 was transfected into BHK-21 cells with electroporation technique. After selection by G418, resistant clones were screened by RT-PCR and Western blotting test. Results：The RT-PCR results of four in five randomly selected cell clones were positive. Western blotting test showed that NS1 gene could be expressed in BHK-21 cells. Conclusions：NS1 protein was capable of being expressed and appropriately processed in pcDNA-NS1 transfected BHK-21 cells. The present results suggest the feasibility of NS1-based DNA immunization.

OBJECTIVE To investigate how MLKL functions on the membrane and explore its electrophysiological characters and structure. METHODS The full-length human MLKL were expressed in SF21 cells and purified using glutathione-sepharose affinity chromatography. The currents of purified MLKL proteins were recorded in avoltage-clamp mode using a Warner BC-535 bilayer clamp amplifier. The currents were digitized using pCLAMP 10.2 software. HEK293 cells were cultured and transfected with MLKL plasmid. Cell viability was examined using the CellTiter- Glo Luminescent Cell Viability Assay kit. RESULT MLKL forms cation channels that are permeable preferentially to Mg2+ rather than Ca2+ in the presence of Na+ and K+. Moreover,each MLKL monomer contains five transmembrane helices:H1, H2, H3 , H5 and H6 of the N-terminal domain which is sufficient to form channels. Finally, MLKL-induced membrane depolarization and cell death exhibit a positive correlation to its channel activity.

Objective To investigate the effect of vascular endothelial growth factor (VEGF) on the differentiation, formation and function of the dendritic cell (DC) in peripheral blood of patients with oral squamous cell carcinoma (OSCC).Methods Flow cytometry was used to detect the number of DC in peripheral blood of 81 patients with OSCC, and ELISA applied to test serum VEGF concentration the OSCC patients, and immunohistochemistry used to observe the expression of VEGF in primary foei of 57 patients with OSCC.DC from CD-14 peripheral blood mononuclear cells were cultured with VEGF165 in vitro to investigate the cytokine's effect on DC.Results In comparison with controls[(325.70±117.54) ng/L], the level of serum VEGF [(764.33±263.64) ng/L] was significantly increased (P < 0.01) and the DC numbers was significantly decreased (P < 0.01) in patients with OSCC.There was a negative correlation between serum VEGF concentration and the level of DC (P < 0.01).The expression of VEGF in primary focus was positively correlated with serum VEGF concentration, but was negatively correlated with the level of peripheral blood DC (P <0.01).DC cultured in vitro with VEGF165 decreased the expression of CD-1a, CD-40,CD-80, CD-86, CD-83, HLA-DR, and revealed a lower ability of stimulating T lymphocyte proliferation but a higher ability of uptake, compared to controls.Conclusions The overexpressed VEGF in patients with OSCC might be one of the important reasons for blocking the differentiation and maturation of DC.

OBJECTIVEIn this study, we try to find the better protocol of limb ischemia postconditioning by observing different protective effects of limb ischemic postconditioning (different strength and time windows in rabbits).

METHODS42 healthy New Zealand rabbits were randomly divided into 7 groups (n = 6): Sham; Control (CON); Skeletal muscle postconditioning (SP); 6 min-delayed skeletal muscle postconditioning (6M-DSP); 1 min-delayed skeletal muscle postconditioning (1M-DSP); Strengthen skeletal muscle postconditioning (SSP); Weakened skeletal muscle postconditioning (WSP). Acute ischemia/reperfusion (I/R) model was induced by 45 minutes occlusion on left circumflex coronary artery (LCX) and 2 hours reperfusion in all anesthetized open-chest rabbits except the Sham. Limb ischemia was induced by external iliac arteries occlusion and reperfusion through artery clamps. The extent of myocardial infarction was assessed by triphenyltetrazolium (TTC) staining. Blood serum creatine kinase (CK) activity and lactate dehydrogenase (LDH) activity were measured at baseline,the end of ischemia, after 1 hour and 2 hours of reperfusion respectively.

RESULTSCompared with the CON, the weight ratio and area ratio of myocardial infarction size were significantly decreased by 49.97% and 43.78% in SP, by 42.32% and 42.68% in 1M-DSP, by 48.36% and 48.86% in SSP (P < 0.05). But there was no significant difference between SP and 1M-DSP and SSP (P > 0.05). Otherwise, compared with the CON, myocardial infarct size was not significantly reduced in 6M-DSP or WSP (P > 0.05). The change of CK was similar to the trend of myocardial infarct size.

CONCLUSIONThe limb ischemia strength of 5 mini/1 minR x 1 cycle could significantly reduce the myocardial ischemia/ reperfusion injury in rabbits, if it was achieved before myocardial reperfusion.

Animals ; Extremities ; blood supply ; Ischemic Postconditioning ; methods ; Male ; Muscle, Skeletal ; blood supply ; Myocardial Infarction ; pathology ; Myocardial Reperfusion Injury ; pathology ; prevention & control ; Myocardium ; metabolism ; Rabbits

OBJECTIVETo evaluate the effectiveness of intravenous immunoglobulin IVIG, 1 g/kg single intravenous injection in treating and preventing cardiac consequences of Kawasaki disease (KD) in children.

METHODSA total of 242 children with KD disease were enrolled in the study. In the randomized controlled trial, they were randomly divided into two groups: IVIG 1 g/kg group and IVIG 2 g/kg group, with aspirin administered within the first 7 to 10 days of illness. The occurrence and restoration of coronary artery lesion (CAL) in these two groups as well as the clinical and laboratory indexes including total fever duration, restoration of cervical lymphadenopathy, white blood cells count, platelet count, serum immunoglobulin, C reactive protein, erythrocyte sedimentation rate and EKG were observed. The clinical effectiveness of the groups before and after the treatment was analyzed.

RESULTSThe age of the 242 children with KD disease ranged from 3 months to 14 years (mean 4.0 +/- 2.8 years old). Male to female ratio was 1.66:1, 83.1% of KD patients were blow 5 years old, 93.4% patients were followed up with echocardiography at the end of the first year and the follow-up period was (38 +/- 18) months, ranging from 4 months to 5.4 years; 86.9% of the cases in 1 g/kg group and 91.7% of the cases in 2 g/kg group had their fever controlled within 48 hours. The difference was not significant (P > 0.05). Serum immunoglobulin level was markedly enhanced after IVIG. Serum immunoglobulin levels in the patients of 2 g/kg group and 1 g/kg group were (26.9 +/- 7.4) g/L and (18.3 +/- 6.9) g/L, respectively (P < 0.01). The average duration of fever in IVIG 1 g/kg group was 10.6 days. After the treatment with 1 g/kg of IVIG, the abnormal white blood cells count, platelet count, C reactive protein, erythrocyte sedimentation rate and abnormal EKG findings were greatly improved (P < 0.001). However, there was no significant difference in the above-mentioned improvement between IVIG 1 g/kg group and IVIG 2 g/kg group (P > 0.05). In IVIG 1 g/kg group the occurrence of CAL was 29.5%. After the one-year follow-up, 87.5% CAL restored, but 12.5% did not, among which 9.4% were those of IVIG non-responders. In IVIG 2 g/kg group the incidence of CAL was 24.2%. After the one-year follow-up, 89.3% CAL restored, but 10.7% did not, all of which were those of IVIG non-responders. There was no significant difference in the incidence of CAL between the two groups (P > 0.05).

CONCLUSIONSingle intravenous injection of IVIG at 1 g/kg could effectively alleviate the clinical symptoms, decrease the incidence of CAL and reduce the complication of cardiovascular system. In the treatment of KD, the therapeutic effectiveness of IVIG at 1 g/kg was not significantly different from that of single intravenous injection of IVIG at 2 g/kg.

Adolescent ; Blood Sedimentation ; C-Reactive Protein ; analysis ; Child ; Child, Preschool ; Coronary Artery Disease ; prevention & control ; Electrocardiography ; Female ; Follow-Up Studies ; Humans ; Immunoglobulins ; blood ; Immunoglobulins, Intravenous ; administration & dosage ; therapeutic use ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; therapy ; Platelet Count ; Treatment Outcome