18 fungal strains including N.coenophialum,N.lolii, N.huerfanum、N.chisosum、N.aotearoae、N.sp.and 8 varieties of grass seeds belonging to Festuca arundinacea and Lolium perenne have been studied.With amplification of IS1～IS3 and F1～R1 of genomic DNA, the primers Tub-2-F～Tub-2-R from Tubulin-2 gene and F3～R3 from NC25 gene have been designed.A PCR method to detect N.coenophialum and N.lolii was established, and also a nested-PCR method to detect N.coenophialum and N.lolii in single seed was established.These PCR detection methods are strongly special and much credible and rapid-speeded.

Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Membrane ; drug effects ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Multiple Myeloma ; drug therapy ; metabolism ; pathology ; Oxides ; pharmacology ; Potassium Channel Blockers ; pharmacology ; Potassium Channels, Voltage-Gated ; drug effects

OBJECTIVETo observe the protection of Qingyuan Shenghua Decoction (QSD) on multiple organs of sepsis patients after bone trauma, and to preliminarily explore its mechanism.

METHODSTotally 60 sepsis patients after bone trauma were randomly assigned to the treatment group and the control group according to random digit table, 30 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took QSD or were nasally fed with QSD, one dose per day for 1 week. Changes of WBC, oxygenation index (PaO2/FiO2), serum creatinine (SCr), total bilirubin (TBIL), aspartate aminotransferase (AST), fibrinogen (FIB), D-dimer (DD), activated partial thromboplastin time (APTT), pro-calcitonin (PCT), C-reactive protein (CRP), heart rate (HR), mean arterial pressure (MAP), intra-abdominal pressure, scores for Acute Physiology and Chronic Health Evaluation II (APACHE II), sequential organ failure assessment (SOFA) scores were observed before treatment and on day 1, 3 and 7 after treatment.

RESULTSCompared with the control group at the same time point, MAP increased at post-treatment day 1 and 3; CRP, APTT, HR, SCr, TBIL, AST, intra-abdominal pressure at post-treatment day 3 obviously decreased in the treatment group (P < 0.05, P < 0.01). WBC, SOFA scores, PCT, CRP, APACHE II, APTT, D-D, HR, SCr, TBIL, AST and intra-abdominal pressure significantly decreased; FIB, MAP and PaO2/FiO2 obviously increased at post-treatment day 7 (P < 0.05, P < 0.01).

CONCLUSIONQSD had good protective effect on multiple organ function in sepsis patients after bone trauma, and its mechanism might be related with effectively clearing endotoxin, alleviating inflammatory reactions, and fighting against coagulation dysfunction.

APACHE ; Blood Coagulation ; Bone Diseases ; complications ; C-Reactive Protein ; metabolism ; Calcitonin ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Inflammation ; Partial Thromboplastin Time ; Protein Precursors ; metabolism ; Sepsis ; drug therapy ; etiology

BACKGROUNDMinimally invasive techniques are gaining wide-spread application in lumbar fusion surgery, because they may have advantage over conventional open surgery in approach-related morbidity. This research was aimed to evaluate the safety and accuracy of the techniques of minimally invasive transforaminal lumbar interbody fusion by using a computer-assisted spinal navigation system combined with electromyography monitoring.

METHODSSixteen patients underwent minimally invasive transforaminal lumbar interbody fusion. A computer-assisted spinal navigation system and electromyography were used for guiding pedicle screw placement. The operative duration, blood loss, complications, and fluoroscopic time were recorded. Clinical outcome was assessed by Visual Analog Scale and Oswestry Disability Index. Radiographic images were obtained to evaluate the accuracy of pedicle screw placement and fusion rates.

RESULTSThe Visual Analog Scale and Oswestry Disability Index scores were vastly improved postoperatively. A total of 64 pedicle screws were implanted and three were regarded as misplacement by post-operative CT scan. Three screw trajectories were adjusted according to intra-operative stimulus-evoked electromyography monitoring. The average fluoroscopy time in each patient was 31.8 seconds, which equals to 7.9 seconds per pedicle screw. No patients had instrument related neurological complications, infection, implant failure or revision. Successful fusion was found in all patients.

CONCLUSIONSThe combination of navigation system and real-time electromyography monitoring can make the minimally invasive operation more safe and accurate while decreasing radiation exposure time of the medical staff and patient and minimizing the chance and the degree of the pedicle screw misplacement.

Adult ; Aged ; Bone Screws ; Electromyography ; methods ; Humans ; Lumbar Vertebrae ; surgery ; Middle Aged ; Monitoring, Intraoperative ; methods ; Spinal Fusion ; methods ; Spine

There are many methods of conservative treatment for early non-traumatic osteonecrosis of femoral head, but still lack of a mature and effective treatment. Gene therapy is a new treatment method for non-traumatic osteonecrosis of femoral head in recent years. This article showed that the non-traumatic osteonecrosis of femoral head related to progress and problems on pathogenesy, molecular biology foundation, exogenous theoretical gene,carrier selection and so on, discussed some problems and countermeasure and future applications.
Femur Head Necrosis ; therapy ; Genetic Therapy ; Humans

OBJECTIVETo investigate the effect of Prostate Water Pellets (PWP) on serum levels of IL-6 and TNF-alpha in rats with chronic bacterial prostatitis (CBP).

METHODSFifty healthy, adult, male Wistar rats with the weight of 180 - 220 g were divided into five groups of ten rats each at random: the control group, model group, high dosage of PWP group, low dosage of PWP group and levofloxacin group. The CBP rat model were created by injecting Escherichia coli (0.2 ml/rat, 10(7)/ml) into prostates. A month later after the model creation, high and low dosage of PWP suspension were used by gavage in CBP rats for 30 days, respectively. Levofloxacin tablets were used by gavage as the positive control, and distilled water was used by gavage in the control and model group. After thirty days, serum levels of IL-6 and TNF-alpha were measured with radioimmunoassay.

RESULTSCompared with the model group, serum levels of IL-6 and TNF-alpha of rats in high and low dosage PWP groups were lower and the difference was significant statistically (P < 0.01).

CONCLUSIONIt has effect to treat CBP rat with the PWP and its mechanism may relate with the decreasing levels of proinflammatory cytokines(IL-6 and TNF-alpha) in blood.

Animals ; Bacterial Infections ; blood ; complications ; drug therapy ; Chronic Disease ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; pharmacology ; Interleukin-6 ; blood ; Male ; Phytotherapy ; Prostatitis ; blood ; drug therapy ; microbiology ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo appraise the effect of single- and two-layer Percoll density gradient centrifugation in sperm separation.

METHODSTwenty semen specimens underwent single-(50%) and two-layer (90% and 45%) density gradient centrifugation, respectively. The sperm class analyzer (SCA) was used to analyze sperm density, motility and dynamic parameters and round cell density before and after the treatment.

RESULTSAfter separation, the sperm recovery rate of the single-layer method was (65.5 +/- 12.8)%, significantly higher than that of the two-layer method (P < 0.01). The percentages of grade a sperm of the single- and two-layer method were significantly higher than pre-treatment (P < 0.05, P < 0.01), that of the single-layer was significantly lower than that of the two-layer method (P < 0.05), but the percentage of grade c sperm of the former was significantly higher than that of the latter (P < 0.05). Compared with pre-treatment, the percentage of grade a + b sperm of the two-layer method was significantly higher (P < 0.05), while that of the single-layer method showed no significant difference (P > 0.05), and the round cell density of both the methods was significantly lower (P < 0.05, P < 0.01), with no significant differences between the two methods (P > 0.05).

CONCLUSIONThe single-layer method yields a higher rate of sperm recovery and causes little change in the sperm motility, while the two-layer method effects a lower rate and significantly improves sperm motility. Both the methods can efficiently separate sperm from round cells, and each has its own advantages and its application value in in vitro treatment of sperm.

Cell Separation ; methods ; Centrifugation, Density Gradient ; methods ; Humans ; Male ; Povidone ; Silicon Dioxide ; Sperm Count ; methods ; Spermatozoa ; cytology

OBJECTIVETo compare the differences of the gene expressions in androgen-independent and androgen-dependent prostate cancer (ADPC), gain a deeper insight into the molecular mechanism of androgen-independent prostate cancer (AIPC), and find effective means for its clinical diagnosis and treatment.

METHODSEats of genes highly-associated with prostate cancer were obtained by mining PubMed with the FACTA tool, and the specifically expressed genes in AIPC were analyzed with a set of bioinformatic tools including GATHER, PANTHER, STRING and ToppGene.

RESULTSA total of 128 genes specifically expressed in AIPC were identified, as compared with 23 that were specific to ADPC. Bioinformatic analysis showed the essential roles of AIPC-specific genes in such important biological processes as cell signal transduction, cell adhesion, apoptosis, oncogenesis, cell proliferation and cell differentiation.

CONCLUSIONSuch genes as MMPJ, EGFR, MMP2, ADM, MIF, IGFBP3, 112, MET, BAD, RHOA, SPP1, EP300, SMAD3, RAE1, PTK2, and TGFB2 may play important roles in transforming ADPC into AIPC.

Androgen Antagonists ; Androgens ; metabolism ; Computational Biology ; Data Mining ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Gene Regulatory Networks ; Genes, Neoplasm ; Humans ; Male ; Prostatic Neoplasms ; genetics ; metabolism

OBJECTIVETo evaluate clinical result of the micro-decompression procedure for lumbar spinal stenosis.

METHODSFrom September 2001 to May 2006,87 patients (male 60, female 27) with lumbar spinal stenosis underwent micro-decompression. The age of patients were from 43 to 80 years with an average of 51 years. Among them,2 cases with spinal stenosis occured in L(3,4), 47 in L(4,5), 38 in L5S1.

RESULTSAll patients were followed up for 18-48 months with an average of 26 months. The results were excellent in 52 cases, good in 28, poor in 7, according to Macnab of back leg pain standard. The rate of excellent and good was 92%.

CONCLUSIONOperative treatment for lumbar spinal stenosis is focused at the areas causing symptomatic neural root compression rather than prophylactic decompression at areas of nonsymptomatic disease. The micro-decompression procedures are more likely to be well tolerated by older patients.

Adult ; Aged ; Aged, 80 and over ; Decompression, Surgical ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Stenosis ; surgery ; Treatment Outcome

Background Biomaterials for corneal tissue engineering must demonstrate several critical features for potentialutility invivo, includingtransparency, mechanicalintegrity, biocompatibilityand slow biodegradation. Silk film biomaterial had been characterized to meet these functional requirements. ObjectiveThis study was to investigate the feasibility of physico-crosslink regenerated silk fibroin film as tissue engineered corneal scaffold. MethodsHuman corneal epithelial cells(CECs) links were cultured by regular method and CECs in logarithmic phase were than incubated on physico-crosslink regenerated silk fibroin film membrane. The shape of cultured human CECs was observed after 24,48 and 72 hours under the inverted microscope and scanning electron microscope( SEM ) ,and the CECs were cultured on culture plates as controls. The growth state of CECs on regenerated silk fibroin film was observed daily for 7 days by MTT, and cell cycle analysis and the presence of apoptosis of human CECs were examined by flow cytometry after incubation on regenerated silk fibroin film. Regenerated silk fibroin filmCECs (4 mm×3 mm) were implanted into the corneal stroma of the right eyes of New Zealand white rabbits. At the end of 4 and 8 weeks after implantation, the appearance of the ocular surface was examined using slit lamp and corneal neovascular area was measured. Corneal histopathological examination was carried out to assess the degradation of graft materials and immunohistochemistry was performed to detect the expression of CD34 in the corneal tissue after operation. ResultsThe morphology and structure of CECs were identical using the two cultured Methods when observed under the inverted microscope and SEM after 24,48 and 72 hours. No significant difference was found in the A490 value 1,2,3,4,5,6 or 7 days after incubation on regenerated silk membrane and in culture plates ( Fmethod =0. 641 ,P＞0.05 ). The apoptosis rates of CECs on regenerated silk membrane or culture plates were 1.8％ and 2.0％ and the amount of cells in G2/G1 phase was 1. 956 and 1. 945, respectively. Histopathological examination showed that the regenerated silk membrane material degraded and was replaced by regular collagen tissue 2 months after implantation,and the presence of neovascular area and inflammatory cells were less prominent in 2 months than 1 month post-implantation. The expression level of CD34 in corneal tissue was evidently lower 1 and 2 months after operation than the Ad-VEGF165-induced positive control group (P＜0. 05), and no significant differences were seen when compared with normal CECs(P＞0.05). ConclusionsPhysico- crosslink regenerated silk fibroin film is an excellent biomaterial for tissue engineered corneal scaffold with good biocompatibility.