Objective:To investigate the analgesic time-effect characteristics and changes in concentrations of rabbit's hypothalamic 5-hydroxytryptamine (5-HT) and noradrenaline (NE) caused by buccal acupuncture in the rheumatoid arthritis (RA) rabbits,and to reveal the analgesic central mechanism of buccal acupuncture,thereby providing a theoretical basis for the treatment of pain by buccal acupuncture.Methods:Forty rabbits were randomly divided into a normal group,a model group,a body acupuncture group,and a buccal acupuncture group,with 10 rabbits in each group.No model was established in the normal group,while equal dose of normal saline was injected at the matched site and time point;rabbits in other groups were subjected to the establishment of RA models using egg protein.From the 27th day of the experiment,rabbits in each group received the designated intervention.Rabbits in the normal group and the model group were fixed for 30 min every day using the same method as those in the other groups.In the acupuncture group,Dubi (ST 35) and Zusanli (ST 36) on bilateral hind limbs were selected.Perpendicular needling (using the needles with 0.25 mm in diameter and 25 mm in length) was performed with twirling manipulation for 15 s at intervals of 5 min.The needles were retained for 30 min and acupuncture was performed once a day.In the buccal acupuncture group,the knee point in the buccal acupuncture and needles with a diameter of 0.25 mm and a length of 15 mm were selected.Oblique needling was performed with twirling manipulation for 15 s at intervals of 5 min.The needles were retained for 30 min and acupuncture was performed once a day.The thermal pain thresholds at the 0,5,15,30,60,120 and 240 min after the 1st and 10th acupuncture therapy were measured with a PL-200 thermal-inducing pain meter.After the 10th acupuncture therapy,rabbit's hypothalamus was removed,and the 5-HT and NE concentrations in the hypothalamus at the peak point of the acupuncture pain threshold curve were determined by high performance liquid chromatography (HPLC).Results:The analgesic effect was obvious at 5 min after buccal acupuncture started,peaked at 30 min,and decreased to the lowest value at 240 min.Rabbits in the body acupuncture group began to show significant analgesic effect at 15 min,which was peaked at 30 min,and began to decline at 60 min.The pain threshold at 240 min was still higher than that at 0 min.Compared with the model group,the concentrations of hypothalamic 5-HT in the buccal acupuncture group and the body acupuncture group was significantly increased,and the between-group differences were statistically significant (both P＜0.05).The NE/5-HT ratios in hypothalamus in the buccal acupuncture group and the body acupuncture group were significantly lower than the ratio in the model group,and the differences were statistically significant (both P＜0.05);difference in the decrease was statistically significant between the buccal acupuncture group and the body acupuncture group (P＜0.05).Conclusion:The analgesic effect of buccal acupuncture shows an obvious time-dependent curve.It is characterized by rapid onset of pain relief,rapid increase and decline in pain threshold.5-HT and NE levels in rabbit's hypothalamus can be affected by buccal acupuncture,with increased 5-HT concentration and reduced NE/5-HT ratio.

There are many E. coli rare codons in the gene of porcine interferon alpha-1. In order to obtain high expression of poIFN-alpha1 in E. coli, the cDNA encoded poIFN-alpha1 mature protein was synthesized using biased codons of E. coli without changing the original amino acid sequence and the terminator was changed as TAA. At the same time, Adenine and Thymine were used to the largest extent near the 5' terminus of poIFN-alpha1 mature protein gene. The synthesized gene was inserted into the Eco RI and Sal I site of the expression vector pRLC resulting pRLC-poIFN-alpha1. The poIFN-alpha1 is highly expressed in E. coli DH5alpha when the induction was carried out at 42 degrees C . The expressed poIFN-alpha1 account for 24.5% of the total cellular proteins and existed as inclusion body. The poIFN-alpha1 inclusion body was dissolved in 6mol/L guanidine chloride contained DTT and subsequently the denatured poIFN-alpha1 was re-natured by dilution in refolding buffer containing GSH and GSSH. In the present study it was found that the denatured poIFN-alpha1 was most efficiently re-natured in refolding buffer containing 1 mol/L guanidine chloride. In order to obtain pure protein, the concentrated re-natured poIFN-alpha1 was purified by Sephacryl S-200 chromatography. As a result, the purified poIFN-alpha1 is verified to be of high cytokine activity by inhibiting the cytopathic effect of vesicular stomatitis virus in MDBK cells, which is about 6.4 x 10(6) u/mg. This study paved the way for large-scale production of recombinant poIFN-alpha1 and its usage in virus disease control of pigs.
Animals ; Codon ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Interferon-alpha ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Swine ; Transduction, Genetic

Objective To discuss the different clinical features and prognosis of single cerebral venous thrombosis (CVT) and multiple CVT. Methods The site and the number of vein and thrombosed sinuses of 136 patients with CVT were summarized. The patients were divided into 2 groups according to the numbers of thrombosed sinuses. The clinical features and outcome of the patients with single CVT were analyzed in comparison with those with multiple CVT by univariate analysis. Results In 44 patients (32.4%), only 1 cerebral sinus was involved. In 92 patients (67.6%), 2 or more cerebral veins and sinuses were involved (2 sinuses in 45, 3 sinuses in 35, 4 sinuses in 9, 5 sinuses in 3). The lateral sinus and the sigmoid sinus were the most frequent thrombosed sinuses which were found in 86.8% of patients; the followings were superior sagittal sinus (58.1%), straight sinus (18.4%) , deep venous system (7.4%), and cortical veins (2.9%). Mean ages were significantly older but the short-term prognosis was better in the group of patients with single CVT in comparison with those in the group of patients with multiple CVT. The patients with multiple CVT also presented more serious intracranial hypertension, more frequent parenchymal lesions and systematic thrombotic events than those with single CVT (P＜0.05). Conclusion In most CVT patients, 2 or more veins and sinuses are involved and thromboses most commonly implicate the lateral sinus and the superior sagittal sinus. Patients with multiple CVT usually present higher intracranial pressure, more serious clinical course, worse outcome and higher incidence of systematic venous thrombotic events in comparison with patients with single CVT. And the multiple sinus thrombosis is more likely to cause venous infarctions and intracranial hemorrhage than the single one.

To enhance the DNA immunogencity of PRRSV ORF5 gene, CpG sequence and the universal helper T cell antigen epitope (PADRE) sequence were inserted between the decoy epitope and the neutralizing epitope. At the same time, site-mutations were introduced at N33 and N51 to diminish the coverage effect to epitope B from the polysaccharides. Subsequently, the modified ORF5 gene (MORF5) and PRRSV ORF6 gene were cloned into the eukaryotic expression vector pcDNA3.0 under the control of two CMV promoters, respectively. With indirect immunofluorescence assay and Western-blot the expression in vitro of the two genes was confirmed, then six-week-old Balb/C mouse were immunized with the modified expression plasmid pcDNA-M5A-6A. The non-modified expression plasmid pcDNA-5A-6A, the blank eukaryotic expression plasmid pcDNA3.0, living attenuated vaccine and inactivated vaccine were used as controls. The PRRSV specific neutralizing antibodies and the T cell proliferation response were elevated with virus neutralization assay and MTf method. Results indicate that the modified plasmid pcDNA-M5A-6A can elicit not only higher titer of neutralizing antibodies in a rapid time, but also more vigorous T cell proliferation response compared with the non-modified plasmid pcDNA-5A-6A and commercial vaccines, indicating that DNA vaccine pcDNA-M5A-6A maybe a promising candidate for PRRS prevention.
Animals ; Antibodies, Neutralizing ; immunology ; Antibodies, Viral ; blood ; immunology ; Binding Sites ; genetics ; Blotting, Western ; CHO Cells ; Cell Proliferation ; Cricetinae ; Cricetulus ; Female ; Glycosylation ; Mice ; Mice, Inbred BALB C ; Mutation ; Open Reading Frames ; genetics ; Porcine Reproductive and Respiratory Syndrome ; immunology ; prevention & control ; Porcine respiratory and reproductive syndrome virus ; genetics ; immunology ; metabolism ; Swine ; virology ; T-Lymphocytes ; cytology ; immunology ; metabolism ; Vaccines, DNA ; administration & dosage ; immunology ; Viral Proteins ; genetics ; immunology ; metabolism ; Viral Vaccines ; administration & dosage ; immunology

In order to obtain a high activity antibacterial peptide, An expression vector pPICZalphaA-pl is constructed with a tandem of four antimicrobial peptides in the same direction,which includes Protegrin-1 (PG-1), Scorpion Defensin (SD), Metalnikowin-2A and Sheep Myeloid Antibacterial Peptide (SMAP-29) (serial number in GenBank are AAB27599, AAAB27538, P80409 and P49928 respectively). At the same time the expression vector pPICZalphaA-sd which express Scorpion Defensin was contructed. The expression vectors of pPICZalphaA-pl and pPICZalphaA-sd were linearized and transformed into the yeast host strain X-33 respectively. Under the control of the promoter AOX1 (alcohol oxidase1), the peptides PL and SD were secreted expressed. Their heat-stable property, acid-stable property and MIC were detected in vitro. The results suggest the peptides PL and SD have good heat-stable and acid-stable properties, and the combinant PL peptide showes higher antibacterial activity against several Gram-positive bacteria (G+) and Gram-negative bacteria (G-) than the peptide SD, especially against Escherichia coli. The antibacterial activity of combinant antimicrobial peptide PL shows its far exploiting perspective.
Animals ; Anti-Infective Agents ; metabolism ; pharmacology ; Antimicrobial Cationic Peptides ; genetics ; pharmacology ; secretion ; Bacillus subtilis ; drug effects ; growth & development ; Blood Proteins ; genetics ; pharmacology ; secretion ; Cathelicidins ; Defensins ; genetics ; pharmacology ; secretion ; Electrophoresis, Polyacrylamide Gel ; Escherichia ; drug effects ; growth & development ; Hydrogen-Ion Concentration ; Pichia ; genetics ; Recombinant Fusion Proteins ; genetics ; pharmacology ; secretion ; Salmonella ; drug effects ; growth & development ; Scorpions ; metabolism ; Sheep ; metabolism ; Staphylococcus aureus ; drug effects ; growth & development ; Time Factors

OBJECTIVETo analyse related risk factors of classic Kaposi' s sarcoma in Xinjiang.

METHODSA 1:4 case-control study was used and the conditional logistic regression model was performed in this study. Cases were followed up in Xinjiang while controls were selected by the same sex, nation and age within 5 years with cases.

RESULTSInterleukin-6,vascular endothelial growth factor, beta-MG, neopterin, human herpevirus 8, were found to be associated with Kaposi's sarcoma risk in one-way variance model while beta2 -MG and human herpevirus 8 entered the multiple conditional logistic regression model, and their ORs were 1.002(95%CI: 1.000-1.003), 81.041 (95%CI: 1.790-3669.620).

CONCLUSIONThere was a correlate relationship between beta2 -MG and classic Kaposi's sarcoma being found that human herpevirus 8 exposure related factors seemed to have played important roles on classic Kaposi's sarcoma in Xinjiang.

Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; China ; Female ; Herpesviridae Infections ; complications ; Humans ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Risk Factors ; Sarcoma, Kaposi ; complications ; epidemiology ; metabolism

OBJECTIVESTo discuss the methods and outcome of shorting proximal femoral and total hip arthroplasty for Crowe IV dysplastic hip of adults.

METHODSFrom July 2000 to February 2006, 13 cases of osteoarthritis secondary to severe development dysplastic hip were treated by total hip replacement and the shorting proximal femoral.

RESULTSThe duration of follow-up ranged from 4 months to 55 months. The average score increased from 36.9 to 84.1 points after the surgery according to Harris. All the patients could walk independently. Their paces were improved obviously and the function of their hips was satisfactory.

CONCLUSIONSThe treatment by total hip arthroplasty and the shorting of posterior femoral is effective and efficient for osteoarthritis secondary to Crowe IV development dysplastic hip in adults. The long-term followup is necessary for further study.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femur ; surgery ; Follow-Up Studies ; Hip Dislocation ; etiology ; surgery ; Humans ; Male ; Middle Aged ; Osteotomy ; methods ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo understand the prevalence of sleep disordered breathing (SDB) in elderly patients with coronary artery disease (CAD) and explore the relations between SDB and CAD.

METHODSSixty-two elderly patients with and 18 without CAD identified by coronary angiography underwent examinations by polysomnography (PSG). Left ventricular ejection fraction (LVEF) was measured by 99Tc equilibrium radionuclide angiography.

RESULTSIn the 62 elderly patients with CAD, 53.2% had SDB, a rate significantly higher that (22.2%) in the 18 non-CAD patients. The CAD patients with SDB had higher respiratory disturbance index (RDI) and body mass index (BMI) and lower arterial saturation of oxygen (SaO2) during sleep, with longer duration of low SPO2 (less that 90%). The incidence of hypertension was higher in CAD patients with SDB than in those without SDB. No significant correlation was found between the severity of coronary artery disease and RDI (r=-0.16, P>0.05).

CONCLUSIONThe elderly patients with CAD have higher incidence of SDB, and appropriate interventions should be administered in those with severe SDB.

Aged ; Aged, 80 and over ; China ; epidemiology ; Coronary Angiography ; Coronary Artery Disease ; complications ; diagnostic imaging ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Polysomnography ; Prevalence ; Sleep Apnea Syndromes ; complications ; epidemiology ; physiopathology

BACKGROUNDLarge animal models with toxin-mediated pancreatic damage have been used extensively in researches with respect to diabetes mellitus and cardiovascular diabetic complications. The present study aimed to establish Chinese Guizhou minipig models with streptozotocin (STZ)-induced diabetes and characterize the animal models by analyzing inflammatory cytokine levels in aortic wall, such as tumor necrosis factor (TNF)-alpha, interleukin-1beta (IL-1beta) and interleukin-6 (IL-6).

METHODSTwenty-two male Chinese Guizhou minipigs (age, 4 to 6 months; weight, 20 kg to 30 kg) were divided into STZ-induced diabetic group (n = 12) and control group (n = 10). STZ (125 mg/kg) was administrated to induce hyperglycemia and afterwards insulin was used to control fasting blood glucose levels below 10 mmol/L. Oral glucose tolerance test (OGTT) was performed before and one month after STZ administration and serum concentrations of alanine transaminase, asparagine transaminase, albumin, blood urea nitrogen, creatinine, lipids and white blood cell count were measured before and six months later. Animals in both groups were euthanized after six months and pancreas was examined immunohistochemically for islet beta cells. Aortic intima of diabetic minipigs and controls was analyzed for TNF-alpha level in tissue conditioned medium by Western blot. TNF-alpha, IL-1beta and IL-6 mRNA levels in aortic intima were assayed by reverse transcription and polymerase chain reaction (RT-PCR).

RESULTSSignificant elevation in serum glucose levels was observed one month and six months after STZ induction (P < 0.001) and markedly increased OGTT values were noted, compared with baseline data. The normal pancreas had many irregular sized islets and small clusters of islet beta cells, while in pancreas of diabetic minipigs islet beta cells almost disappeared. No statistical difference was notified in serum concentrations of biochemical examinations before and six months after STZ induction. Western blot demonstrated dramatically increased TNF-alpha level in aotic intima conditioned medium, and significant elevation of TNF-alpha, IL-1beta and IL-6 mRNA levels was revealed by RT-PCR.

CONCLUSIONSThe present study has established Chinese Guizhou minipig models with STZ-induced diabetes. Inflammatory cytokines (TNF-alpha, IL-1beta and IL-6) significantly elevated in aortic intima of diabetic minipigs.

Animals ; Aorta ; chemistry ; Diabetes Mellitus, Experimental ; immunology ; pathology ; Glucose Tolerance Test ; Immunohistochemistry ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Male ; Pancreas ; pathology ; Streptozocin ; Swine ; Swine, Miniature ; Tumor Necrosis Factor-alpha ; blood

Intestinal Peyer's patches （PPs） are local immune tissues of the intestine， which are considered to be the main induction site of the intestinal mucosal immune response， and closely related to immune-related refractory enteropathies， such as ulcerative colitis and Crohn's disease. In recent years， more and more scholars have tried to find a new breakthrough for treating refractory enteropathies with a limited efficacy of clinical interventions through in-depth study of the relationship between PPs and enteropathy. Traditional Chinese medicine （TCM） polysaccharides are considered to be a key component for immune regulation with TCM. Modern studies show that TCM polysaccharides have a significant positive intervention effect on the structure and function of PPs， with good development prospects. Based on this， this paper focuses on PPs and intestinal-related diseases， and systematically introduces the physiological structure of PPs and their drug delivery mechanism， and summarizes the interactions of PPs with effect on immune-related enteropathies， analyze of current studies and prospects of effect of TCM polysaccharides in intervening intestinal disease and its dysfunction by regulating PPs， with the aim to provide new strategies for basic studies and clinical treatment of immune-related refractory enteropathies from the perspective of PPs， and new ideas for basic studies and clinical studies on effect of TCM polysaccharides in intervening enteropathies.