Objective: To construct eukaryotic expression vector pSecTag2/HygroB-DAF of human decay accelerating factor (DAF) and transfect NIH/3T3 cells after encapsulated by chitosan. Methods:The human DAF fragments were obtained by PCR form DAF-pGEM-T Easy Vector, cloned into the eukaryotic expression vector pSecTag2/HygroB, and identified by restriction endonuclease’s digestion and DNA sequencing. After the particles of pSecTag2/HygroB-DAF were encapsulated by chitosan, the NIH/3T3 cells were transfected by chitosan-DAF nanoparticles and detected DAF expression by immunohistochemistry stain. Results:The DAF fragment was 1049 bp. Its sequence was as same as DAF cDNA in Genebank. After having been transfected by chitosan-DAF nanoparticles 24 hours, the NIH/3T3 cells showed diffusely positive in cytoplasms by anti-DAF immunohistochemistry. Conclusion:Eukaryotic expression vector of human DAF were constructed successfully and transfected it to NIH/3T3 cells after encapsulated by chitosan.

OBJECTIVETo study the anti-endotoxin of different concentration baicalin.

METHODS6.250 microg/mL, 3.125 microLg/mL, 1.562 microg/mL and 0.781 microg/mL baicalin solutions were mixed with I EU/mL endotoxin, respectively. The mixtures were put into water of (37+/-1) degrees C for 15 min, 30 min and 60 min. The degrading effects were determined by using limulus amebocyte lysate test (LAL test).

RESULTS1) The degrading effect of 6.250 microg/mL, 3.125 microg/mL and 1.562 microg/mL baicalin solution on I EU/mL endotoxin was degraded completely in 15 min, 30 min and 60 min, respectively. 2)The degrading effect of 3.125 microg/mL, 1.562 microg/mL and 0.781 microg/mL baicalin solution on 1 EU/mL endotoxin after these mixtures had been incubated for 15 min. Endotoxin values were (0.155 5 +/- 0.002 8) EU/mL, (0.212 1+/-0.004 9) EU/mL, (0.355 9+/-0.013 9) EU/mL, respectively. These differences among them were statistically significant (P<0.01). 3) The degrading effect of 1.562 microg/mL and 0.781 microg/mL baicalin solution on 1 EU/mL endotoxin after these mixtures had been incubated for 30 min. Endotoxin values were (0.1640+/-0.0025) EU/mL and (0.2094+/-0.004 4) EU/mL, respectively. These differences between them were statistically significant (P<0.01).

CONCLUSIONThe action of anti-endotoxin of baicalin is dose-dependent and time-dependent. The results show that baicalin has the stronger anti-endotoxin effect.

Endotoxins ; Flavonoids ; Limulus Test

Objective To investigate the effects of various degrees of sleep deprivation on cognitive and cerebral mitochondrial respiratory function in rats. Methods Sprague-Dawley rats were subjected to a 10-day sleep deprivation protocol and were divided into 5 groups randomly: allowing sleep for 12 h (control group), 9 h (group1), 6 h( group2), 3 h (group3), 0 h each day (group4). The modified multiple platform method(MMPM) was used to establish sleep deprivation model. The cognitive function was tested by Y-type maze. Mitochondrial respiratory function was measured by Clarktype oxygen electrode. Results The number of error reactions was significantly increased in sleep deprived rats. There were statistically significant differences( P ＜ 0.05 )in the number of error reactions between the control group and the completely deprived(0 h group)or severely deprived(3 h group)rats on each of the study days. The oxygen consumption rate in state 3 respiration was significantly decreased( P ＜ 0.05 )in sleep deprived rats compared with the control group. The oxygen consumption rate in state 4 respiration was significantly decreased(P ＜ 0.05 )only in the completely deprived(0 h group)rats compared with the control group. Conclusion Sleep deprivation, no matter mild or severe, both could have adverse effects on cognitive function and cerebral mitochondrial respiratory function.

AIMTo observe the effect of FAK-related non-Kinase (FRNK) plasmid on hepatic stellate cell (HSC) proliferation stimulated by fibronectin (FN).

METHODSFRNK plasmid was transfected into HSC with transient liposomal transfection. The proteins of FRNK, FAK and p-FAK(Tyr397) were assayed by Western blotting analysis. The proliferation of HSC was evaluated by improved MTT assay, and cell cycle pattern was determined by flow cytometry (FCM).

RESULTS(1) The expression of FRNK protein increased after FRNK transfected HSC, and it was at 48 h that the expression of FRNK protein was the highest (P < 0.01). The protein level of FAK was no significant difference between before FRNK plasmid transfection and after transfection (P > 0.05). The expression of p-FAK(Tyr397) protein was down-regulated after FRNK had been transfected in HSC, (P < 0.01). (2) The HSC proliferation inhibition rates at 12 h, 24 h and 48 h after FRNK transfection were 20.07%, 26.16%, 29.77%, respectively (P < 0.01). (3) Compared with the non-FRNK plasmid group, the FRNK-transfected HSCs almost arrested in G0/G1 phase (71.4 +/- 2.81 vs 48.9 +/- 1.66, P < 0.01).

CONCLUSIONAfter FRNK were transfected successfully in HSCs using lipofectamine, the phosphorylation of FAK was inhibited. The HSC proliferation was restrained in a time-dependent manner and the HSC was arrested in G0/G1 phase.

Cell Line ; Cell Proliferation ; Fibronectins ; Hepatic Stellate Cells ; cytology ; Humans ; Phosphorylation ; Plasmids ; genetics ; Protein-Tyrosine Kinases ; genetics ; Transfection

OBJECTIVEPatients with typical clinical manifestations of Hemorrhagic fever with renal syndrome (HFRS) are becoming fewer. We conducted analysis on clinical features of HFRS in order to reduce the mistakes in diagnosis.

METHODS64 patients were diagnosed as HFRS during May, 2000 to June, 2006 in our hospital. All the patients' serological tests (HFRS-NP-specific IgM, IgG antibody) by ELISA method were positive. We collected their clinical manifestations and test results. SPSS 12.0 was used in our statistical analysis.

RESULTSAmong the 64 patients, 71.6% of all the cases occurred from Feb. to June. Most of patients were admitted to the hospital with untypical manifestation. Only 30.6% patients appeared headache, lumbago, and pain of orbital cavity. 32.8% patients had obviously signs of injection and hemorrhage. However, there were 90.6% patients with headache and 84.4% patients with nausea or vomit. Hypotensive or oliguric phases were absent in 56.3% patients. There were only 31.3% patients with all five stages. Thrombocytopenia (79.7%) and heavy proteinuria (71.9%) were common. But 54.7% of patients shown normal or even decreased white blood cell count. Only 2/3 of patients had elevated serum creatinine (Cr). Liver involved was common showing as elevated aminotransferase. ALT level was not always parallel to Cr level. There was an opposite trend between them.

CONCLUSIONWe must recognized the untypical manifestations of HFRS. Further study focus on pathogenesis was useful for diagnosis and therapy.

Adolescent ; Adult ; Aged ; Alanine Transaminase ; blood ; Creatinine ; blood ; Female ; Hemorrhagic Fever with Renal Syndrome ; blood ; pathology ; physiopathology ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Male ; Middle Aged

OBJECTIVETo analyze the mid-term clinical and radiographic results obtained with the Bernese periacetabular osteotomy for the treatment of severe hip dysplasia.

METHODSFrom October 1997 to December 2002 20 hips of 18 patients were classified as having severe acetabular dysplasia (Severin classification Grade IVb). Preoperatively, all patients had hip pain, and sufficient hip joint congruency on functional radiographs. All 20 hips underwent Bernese periacetabular osteotomy. Postoperatively, the hips were assessed radiographically on center edge angle (CE), acetabular roof obliquity and the progression of osteoarthritis. Clinical results and hip function were measured with the Harris hip score at an average of 6.2 years follow-up.

RESULTSComparison of preoperative and follow-up radiographs demonstrated significant improvements in the lateral CE angle, the anterior CE angle, and roof obliquity. The average Harris hip score improved from 78.5 points preoperatively to 91.1 points at the time of the latest follow-up. Fourteen of 18 patients were satisfied with the result of the surgery, and 16 of 20 hips had a good or excellent clinical result. Under-correction occurred in 5 hips.

CONCLUSIONSThe Bernese periacetabular osteotomy is an effective procedure for surgical correction of the severe dysplastic hip. This osteotomy can predictably obtain major reorientation of the acetabulum in all planes. The clinical results in the mid-term follow-up are encouraging.

Acetabulum ; surgery ; Adolescent ; Adult ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Humans ; Male ; Osteotomy ; methods ; Treatment Outcome ; Young Adult

BACKGROUNDIntratracheal instillation of blood induces self-repaired acute lung injury. However, the mechanism of repair has been unclear. Heme-oxygenase (HO)-1, which catalyzes heme breakdown, acts as an inducible defense against oxidative stress and plays an important role in inflammation. The objective of this study was to test the role of HO-1 in lung injury caused by intratracheal instillation of red cells.

METHODSForty healthy, male Sprague-Dawley rats were randomly divided into five groups: normal group, saline group, erythrocyte group, erythrocyte+zinc-protoporphyrin (ZnPP, HO-1 inhibitor) group and saline+ZnPP group. At 2 days after intratracheal instillation of red cells, lung tissues and lavage samples were isolated for biochemical determinations and histological measurements.

RESULTSHistological analysis revealed that administration of ZnPP worsened the acute lung injury induced by instilled erythrocytes. HO-1 was over-expressed in the erythrocyte group and in the erythrocyte + ZnPP group. Compared with the erythrocyte + ZnPP group, the levels of total protein, lactate dehydrogenase and tumor necrosis factor-alpha in the lavage were lower (P < 0.01), while the level of interleukin-10 was higher in the erythrocyte group (P < 0.01).

CONCLUSIONHO-1 protects against erythrocyte-induced inflammatory injury in lung.

Animals ; Erythrocytes ; physiology ; Heme Oxygenase (Decyclizing) ; analysis ; physiology ; Interleukin-10 ; analysis ; Lung ; pathology ; Lung Injury ; prevention & control ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; analysis

The objective of the study is to find out the effect of shark chondroitin on T lymphocyte subsets in cancer patients. Patients were divided into two groups. One group was treated with chemotherapy alone, and the other group was treated with chemotherapy plus shark chondroitin. Using immunofluorescence technique, T lymphocyte subsets in the peripheral blood were determined in two groups before and after chemotherapy. The results showed that CD4(+)/CD8(+) ratio increased in the patients received shark chondroitin. In the chemotherapy group, CD3(+) had no change, but CD4(+) decreased while CD8(+) increased significantly. The results suggest that shark chondroitin could enhance immune function in cancer patients, especially during chemotherapy.

Adenine ; analogs & derivatives ; pharmacology ; therapeutic use ; Adult ; Drug Resistance, Viral ; Female ; Genes, Viral ; Genotype ; Hepatitis B virus ; drug effects ; genetics ; Hepatitis B, Chronic ; drug therapy ; Humans ; Male ; Middle Aged ; Organophosphonates ; pharmacology ; therapeutic use

Objective To study the metabolic changes of auditory cortex in patients with presbycusis by using proton magnetic resonance spectroscopy(1H-MRS). Methods Ten normal hearing volunteers(youth group),10 normal hearing of elderly (aged group) and 8 patients with presbycusis (presbycusis group ) were checked with proton magnetic resonance spectroscopy. N-acetylaspartic acid ( NAA),creatine ( Cr),choline ( Cho),γ-aminobutyric acid (GABA),glutamic acid ( Glu ) compound were measured.The differences between the groups were semi-quantitatively analyzed.Results When compared with youth group,reduced NAA/Cr,increased Cho/Cr were found in the aged group and presbycusis group (P ＜ 0.05 ). GABA/Cr ratio and Glu/Cr ratio were significant difference between presbycusis group and youth group(P ＜ 0.05 ). There were no significant difference in the GABA/Cr and Glu/Cr ratios in the bilateral auditory cortex between the youth group and the aged group(P ＞ 0.05 ).When compared with aged group,the metabolic changes of auditory cortex in patients with presbycusis were remarkable ( P ＜ 0.05 ).Conclusions 1H-MRS is a noninvasive technique that can provide useful information concerning the metabolic changes of auditory cortex in human. In comparison to the aged group and the youth group,the changes of NAA,GABA,Cho and Glu is found in auditory cortex in patients with presbycusis.