Administration, Intranasal ; Humans ; Mucociliary Clearance ; drug effects ; Nasal Mucosa ; drug effects

OBJECTIVEThe purpose of this study was to develop RT- PCR assay for Rapidly detect and distinguish between Norovirus genogroup I and genogroup II with a pair of primers.

METHODSA pairs of primers specific to capsid prote in ORF2 gene of G I and G II Norovirus were dsigned according to the published complete genome sequence, with which the RNA of Norovirus was extracted and RT-PCR amplification. The sensitivity, specificity of the RT- PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.

RESULTSThe results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses, including rotavirus, enteric adenovirus and hepatitis A virus. The detection limit of RT-PCR assay for Norovirus G I and G II were up to 100 pg/ml and 10 pg/ml respectively.

CONCLUSIONThe RT- PCR assay provide rapid and sensitive detection of Norovirus G I and G II and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.

Caliciviridae Infections ; diagnosis ; virology ; DNA Primers ; genetics ; Gastroenteritis ; diagnosis ; virology ; Humans ; Norovirus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods

Animals ; Esophagus ; metabolism ; Glucagon-Like Peptide 2 ; pharmacology ; Intestinal Mucosa ; drug effects ; metabolism ; Intestine, Small ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Stomach ; metabolism

OBJECTIVETo investigate the advantages and disadvantages of unilateral pedicle screw fixation combined with contralateral translaminar facet screw fixation and interbody fusion with cages in the treatment of two-level lumbar vertebra diseases, by comparing bilateral pedicle screw fixation and interbody fusion with cages.

METHODSForty-nine patients with two-level lumbar diseases who received treatments from June 2009 to December 2011 were included in this study. Among these patients, 23 patients received unilateral pedicle screw fixation combined with contralateral translaminar facet screw fixation and interbody fusion with cages (combined fixation group) and the remaining 26 patients underwent bilateral pedicle screw fixation and interbody fusion with cages (bilateral fixation group). These patients consisted of 17 males and 32 females, ranging in age from 29 to 68 years old. Among these patients, lumbar intervertebral disc herniation accompanied by the spinal canal stenosis was found in 29 patients, degenerative lumbar disc diseases in 17 patients and lumbar degenerative spondylolisthesis (degree I) in 3 patients. The lesions occurred at L2,3 and L3,4 segments in 1 patient, at L3,4 and L4,5 segments in 30 patients, and at L4,5 segment and L5S1 segment in 18 patients. Wound length, operation time, intraoperative blood loss and postoperative wound drainage were compared between two groups. Intervertebral space height in the lesioned segment before and during surgery and at the latest follow up was also compared between two groups. Before surgery and at the latest follow-up, the Cobb angle of the coronal plane and sagittal plane of the lumbar spine, loosening or breakage of internal fixations, the dislocation of intervertebral cages, and interbody fusion were all evaluated in each group. The visual analogue scale (VAS) was used to measure lumbar incision pain. The Japanese Orthopedic Association (JOA) scoring system was used to evaluate the function before surgery and at the latest follow-up.

RESULTSNo wound infection or skin necrosis was observed after surgery in all patients. No cerebrospinal fluid leakage, nerve root injury, cauda equia injury or worsened neural function in the lower limb occurred in all patients during and after surgery. Wound length, operation time, intraoperative blood loss and postoperative wound drainage in the combined fixation group were superior to those in the bilateral fixation group. At postoperative 72 hours, the VAS score in the combined fixation group (1 to 4 points, mean 2.35±1.20) was significantly lower than that in the bilateral fixation group (2 to 5 points, mean 3.11±1.00; P<0.05). All the patients were followed up for 12 to 48 months, with a mean of 29 months. After surgery, intervertebral space height was well recovered in each patient and it was well maintained at the latest follow-up, and there was no significant difference between two groups (P>0.05). During follow-up, pedicle screw and translaminar facet screw loosening, dislocation or breakage and dislocation of intervertebral cages were all not found. At the latest follow-up, the Cobb angle of the coronal plane and sagittal plane of the lumbar spine was obviously improved and was not significantly different between two groups (P>0.05). The lumbar interbody fusion rate was 93.5% and 96.2% in the combined fixation group and bilateral fixation group, respectively, and there was no significant difference between them (P>0.05). There was a significant difference in JOA score between before surgery and at the latest follow-up in each patient (P<0.05), and at the latest follow-up, significant difference in JOA score was found between two groups (P<0.05).

CONCLUSIONCompared to bilateral pedicle screw fixation and lumbar interbody fusion with cages, unilateral pedicle screw fixation combined with contralateral translaminar facet screw fixation and lumbar interbody fusion with cages shows advantages including small skin incision, minimal invasion, ease of operation, highly reliable stability, high interbody fusion rate, rapid recovery in the treatment of two-level lumbar vertebra diseases and therefore can be preferred as a treatment method of this disease.

Adult ; Aged ; Female ; Humans ; Intervertebral Disc Degeneration ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Pedicle Screws ; Spinal Fusion ; methods ; Spinal Stenosis ; surgery ; Spondylolisthesis ; surgery

OBJECTIVETo study the maturation effect of CpG2006 and phosphodiester oligonucleotides on leukemia-derived dendritic cells.

METHODSLeukemia cells K562/A02 were induced into dendritic cells by rhGM-CSF and rhIL-4. After 7 days induction, the cell-morphology was observed, the immunophenotype of cells was detected by flow cytometry and the cell function was evaluated by allogeneic mixed lymphocyte reactions, CTL responses and secretion of IL-12 and IL-6. Then a CpG oligonucleotide CpG2006, two synthetic bacterial phosphodiester oligonucleotides A-ODN and T-ODN were added to these leukemia-derived DCs. Three days later, the DCs were re-detected by the above-mentioned methods.

RESULTSAfter induced by CpG2006, A-ODN or T-ODN, the leukemia-derived DCs with typical dendritic morphology were increased. The expressions of CD83, HLA-DR and CD86 were (65.5 +/- 8.4)%, (32.0 +/- 4.3)% and (18.6 +/- 3.2)% respectively in day 7 leukemia-derived DCs, raised to (88.9 +/- 3.6)%, (53.9 +/- 3.2)% and (39.9 +/- 7.3)% respectively after exposing CpG2006 for 3 days; increased to (97.0 +/- 5.3)%, (63.9 +/- 7.3)% and (40.2 +/- 7.4)% respectively after treated by A-ODN; and further increased to (93.26 +/- 4.65)%, (58.3 +/- 5.6)% and (36.2 +/- 6.8)% respectively after treated by T-ODN. These results was markedly different than unaffected cells did. These DCs induced by the above-mentioned three oligonucleotides could upregulate significantly the capacity for stimulating allogeneic T cells. They could also induce CTL to generate specific cytotoxic activity against K562/A02 cells. The secretion of IL-6 and IL-12 was increased remarkably.

CONCLUSIONCpG2006, as well as two phosphodiester oligonucleotides can induce leukemia-derived DCs maturation.

Cell Differentiation ; drug effects ; Cell Survival ; drug effects ; Dendritic Cells ; cytology ; drug effects ; Humans ; K562 Cells ; Oligodeoxyribonucleotides ; pharmacology ; Oligonucleotides ; pharmacology

OBJECTIVETo evaluate the relationship between allergic symptoms and retinoic acid-related orphan receptor variant 2 (RORC2) and interleukin (IL) 17 in patients with allergic rhinitis (AR).

METHODSBlood sample, nasal secretion and nasal mucosa were taken from 23 patients with AR and 16 health individuals. The expression of RORC2 and IL-17 were detected by immunohistochemistry and quantitative real-time fluorescence reverse polymerase chain reaction. The allergic symptoms in patients were graded.

RESULTSThe rate of positive cells of RORC2 and IL-17 in AR group were 0.17 ± 0.05 and 0.72 ± 0.13, higher than the 0.05 ± 0.02 and 0.27 ± 0.11 of health controls, the difference was statistically significant (t were 9.51 and 11.92 respectively, all P < 0.05). The expression level of RORC2 mRNA in nasal mucosa and peripheral blood of AR group were 0.063 ± 0.011 and 0.452 ± 0.031, higher than the 0.029 ± 0.009 and 0.239 ± 0.027 of health controls, the difference was statistically significant (t were 6.51 and 3.35 respectively, all P < 0.05). The concentrations of IL-17 in the nasal mucosa, nasal secretions and serum levels of AR group were (70.28 ± 10.69), (45.32 ± 8.55) and (6.76 ± 1.18) pg/ml, compared with (18.43 ± 8.34), (6.83 ± 1.31) and (0.74 ± 0.05) pg/ml of controls, the difference was statistically significant (t were 7.92, 17.66 and 15.43 respectively, all P < 0.05). The allergy symptom scores of AR group were 9.43 ± 1.27. There were correlations between the allergic symptom and the expression of RORC2 mRNA and IL-17 in nasal mucosa and peripheral blood (r value were 0.820, 0.746, 0.629, 0.841 respectively, all P < 0.05).

CONCLUSIONRORC2 and IL-17 involved in the inflammatory response of AR and can be used as an indicator to judge the severity.

Adult ; Case-Control Studies ; Female ; Humans ; Inflammation ; Interleukin-17 ; metabolism ; Male ; Middle Aged ; Nasal Mucosa ; metabolism ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; metabolism ; Rhinitis, Allergic, Perennial ; metabolism ; pathology ; Rhinitis, Allergic, Seasonal ; metabolism ; pathology ; Young Adult

OBJECTIVETo study the biotransformation of podophyllotoxin by the cell suspension culture and root culture systems of Rheum palmatum.

METHODUsing plant tissue culture technology and HPLC techniques to isolate products. The structures were elucidated by spectroscopic means.

RESULTCell suspension culture of R. palmatum could convert podophyllotoxin to produce picropodophyllotoxin with the yield of 73.8%, while root culture of R. palmatum could convert podophyllotoxin to produce epipodophyllotoxin and apopodophyllotoxin.

CONCLUSIONPodophyllotoxin did not affect the pH value of the media used in tissue cultures. Both cell suspension culture and root culture of R. palmatum can convert podophyllotoxin.

Chromatography, High Pressure Liquid ; Molecular Structure ; Plant Roots ; metabolism ; Podophyllotoxin ; chemistry ; metabolism ; Rheum ; metabolism ; Tissue Culture Techniques ; methods

OBJECTIVETo investigate the effects of the expression of the PPAR-gamma gene on the proliferation and glycolysis metabolism of prostate cancer cells.

METHODSUsing RNAi, we constructed lowly--expressed shRNA-PPARgamma adenoviruses and transfected them to PC3 prostate cancer cells, with blank vectors as controls. Then we detected the proliferation and apoptosis of the cells, glycolysis metabolism related genes and lactate accumulation by CCK-8 kit, and compared the results between the two groups.

RESULTSCompared with the control group, the PPAR-gamma gene expression was obviously inhibited by RNAi in the PC3 cells, and its protein expression was reduced to (26.00 +/- 4.06)%. The proliferation inhibition rate was (39.5 +/- 4.92)% on the 2nd day, and the apoptosis rate was as high as (21.03 +/- 3.08)%. The glycolysis metabolism related gene products (Myc and Glut-1) were significantly decreased, and the lactate concentration was reduced to 69.71% of that of the controls on the 4th day. There were statistically significant differences in the above findings as compared with the control group (P < 0.01).

CONCLUSIONPPAR-gamma gene knockdown is expected to be a new way to treat prostate cancer.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Glucose Transporter Type 1 ; metabolism ; Glycolysis ; Humans ; Male ; PPAR gamma ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-myc ; metabolism ; RNA Interference ; RNA, Small Interfering ; Transfection

OBJECTIVETo study the effects of Salvia miltiorrhiza on treatment of Chlamydia trachomatis salpingitis (CTS) and fibrosis.

METHODA mouse model for CTS was estahlished in C3H/He by intravaginal inoculation. after 3 weeks mice were randomly divided into 3 groups. Only Azithromyxin was given orally, Azithromyxin and early S. miltiorrhiza given, or Azithromyxin and later S. miltiorrhiza given. After 10 weeks, observe the change of oviduct of mice, observe the histopathologic change and analysis collagen histochemical index.

RESULT3 Treatment groups induce tubal occlusion and hydrosalpinx decreased and the collagen histochemical index decreased significantly than those of no treatment given (P < 0.05). Early S. miltiorrhiza given group induce tubal occlusion and hydrosalpinx decreased and the collagen histochemical index decreased significantly than only Azithromyxin group or later S. miltiorrhiza given group (P <0.05).

CONCLUSIONWhen we treat CTS genital infection with Azithromyxin, if we can give S. miltiorrhiza treatment as early as possible, it may decrease tubal occlusion and hydrosalpinx. significantly inhibit fibrosis maybe one of its pharmacologic mechanismin.

Animals ; Chlamydia Infections ; complications ; drug therapy ; microbiology ; Chlamydia trachomatis ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Fallopian Tube Diseases ; etiology ; prevention & control ; Fallopian Tubes ; drug effects ; pathology ; Female ; Fibrosis ; Mice ; Mice, Inbred C3H ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Salpingitis ; complications ; drug therapy ; Salvia miltiorrhiza ; chemistry

OBJECTIVESTo explore the relationships between the peripheral blood levels of CD61, CD63, PAC-1 and the incidence of acute rejection and tubular necrosis after renal transplantation, and recovery of the graft function.

METHODSThe peripheral blood levels of CD61, CD63, and PAC-1 of 86 patients with uremia in different stages before and after transplantations were analyzed by flow cytometry. The patients were divided into three groups: (1) twenty-nine patients with normal grafts function, (2) hirty with acute rejection and (3) twenty-seven with acute tubular necrosis. The patients with acute rejection were randomly divided into treatment group with anticoagulants and cntrol group.

RESULTSThe peripheral blood levels of CD61, CD63 and PAC-1 significantly increased (P < 0.05) in the patients with acute rejection, in comparison with those with normal grafts function and those with acute tubular necrosis. The peripheral blood levels of CD61, CD63 and PAC-1 in patients with acute rejection in anticoagulants therapy was lower, recovery time of the grafts function was shorter, one-year survival rates of patients and grafts were higher, as compared with those of controls.

CONCLUSIONSThe patients with acute rejection have significantly high peripheral blood levels of CD61, CD63 and PAC-1 before transplantation, however, these values in patients with acute tubular necrosis are not high, this suggesting that acute rejection might relate to platelet activation, while acute tubular necrosis might not relate to it. After anticoagulants therapy in patients with acute rejection, the grafts function might recover faster and their one-year survival rates and grafts might be higher in those with CD61, CD63 and PAC-1 decreasing remarkably.

Adult ; Aged ; Antigens, CD ; blood ; Dual Specificity Phosphatase 2 ; Female ; Graft Rejection ; Humans ; Integrin beta3 ; blood ; Kidney ; physiopathology ; Kidney Transplantation ; Male ; Middle Aged ; Platelet Activation ; Platelet Membrane Glycoproteins ; Protein Phosphatase 2 ; Protein Tyrosine Phosphatases ; blood ; Tetraspanin 30