A reverse transcription polymerase chain reaction (RT-PCR) and single-strand conformation polymorphisms (SSCP) assay were applied to rapidly detect the molecular variability in CP and SP genes among seven isolates of Rice stripe virus in China. The PCR results showed that the CP gene of JD isolate and SP gene of PJ isolate could not be amplified. SSCP analysis showed that there were completely different electrophoretic pattern of CP gene among six isolates. To SP gene, SSCP results also discovered polymorphisms. There were five patterns among these isolates, and the pattern of YL and BS isolates were same.

Adolescent ; Adult ; Aged ; Female ; Humans ; Joint Dislocations ; physiopathology ; therapy ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Shoulder Joint ; injuries ; physiopathology ; Treatment Outcome ; Young Adult

The technique conditions of decolonization of fermentation broth were successively optimized using single factor assay and orthogonal layout method in Cordyceps jiangxiensis. The optimal condition of decolorization was investigated to be 3g/100mL active carbon, 5 min absorption time, pH5 of fermented broth and 25℃absorption temperature. Under the optimal condition, the maximum decolorization rate of fermented broth reached 89. 6% , simultaneously 10. 7% consuming rate of exopolysaccahride was minimum. Subsequently, the extract condition of exopolysaccharide of C. jiangxiensis was further optimized by orthogonal layout design. The maximum exopolysaccharide production was 0. 38 g/L under the optimal condition, i. e. firstly fermented filtrate decolorized and deproteined was concentrated to 1/7 of its total volume, secondly concentration broth was mixed with four times its volume of absolute ethanol and stirred vigorously, lastly precipitation of exopolysaccharide proceeded at 4℃for 16 hrs and the exopolysaccharide collected by centrifugal ion and dryness.

Objective To explore the importance and methods of retaining articularis during pri- mary total hip arthroplasty(THA)and reconstruct soft tissue balance of hip joint after THA.Methods From February 2003 to August 2005,41 eases(43 hips)including 19 males and 22 females at age of 46- 80 years(mean 66.5 years)were treated with THA with retained capsule(Group R)and other 42 cases (44 hips)including 20 males and 22 females at age of 43-80 years(mean 64.3 years)with standard THA (Group S).Preoperative diagnosis found femoral neck fractures(GardenⅢⅣ)in 13 cases(13 hips)in Group R and 14(14 hips)in Group S;acetabular dysplasia(CroweⅢ)in 9(9 hips)in Group R and 8 (hips)in Group S;Osteoarthritis in 6(8 hips)in Group R and 7(8 hips)in Group S;and femoral head osteonecrosis(FicatⅢⅣ)in 13(13 hips)in Group R and 13(14 hips)in Group S.There were 13 hips of cement prostheses in Group R and 11 in Group S,8 cementless prostheses in Group R and 8 in Group S, 22 cement and cementless prostheses in Group R and 23 in Group S.Gibson's approach was used in both groups.Group R used the method of retaining capsule and little supination muscles during the operation to reconstruct responsibly soft tissue balancing of postoperation for THA.For comparison,Group S used the method of standard which resected a lots of capsule and didn't reconstruct it.The comparative items between Group R and Group S included incisional length,operative time,operative bleeding,drainage transfusion, infection,dislocation,postoperation standing,postoperation walking and Harris's score.Results All cases in Group R and Group S were followed for 6-22 months(mean 16.5 months in Group R and 16.7 months in Group S).There was significantly statistical difference upon interoperative and postoperative data between Group R and Group S.The result of Group R was significantly better than that of GS.Conclu- sion Retaining articularis during primary THA can minimize operative trauma,reconstruct soft tissue bal- ance and augment hip stability to get postoperative functional recovery.

Cell Line ; Cytochrome P-450 CYP1A1 ; drug effects ; metabolism ; Hepatocytes ; cytology ; metabolism ; Humans ; Microsomes, Liver ; drug effects ; metabolism ; Quercetin ; pharmacology

OBJECTIVETo explore the clinical application of the tubularized incised plate (TIP) in the surgical treatment of hypospadias.

METHODSThis study included 169 cases of hypospadias treated by TIP surgery from January 2007 to April 2009. The patients ranged in age from 1.5 to 12 years (mean 3.68 yr). The TIP technique was modified based on that described by Snodgrass, with the urethral plate longitudinally incised and a urethral stent kept in place. The patients were hospitalized for 10 days postoperatively, and followed up for an average of 2 years, ranging from 6 months to 3 years.

RESULTSComplications developed in 18 (10.6%) of the patients, most frequently meatal stenosis (9 cases, 5.3%) and urethrocutaneous fistula (8 cases, 4.7%).

CONCLUSIONThe TIP technique, as a surgical method, can be applied to most hypospadias cases. The accumulation of clinical experience and skills may help raise the success rate and reduce the complications of TIP surgery.

Child ; Child, Preschool ; Humans ; Hypospadias ; surgery ; Infant ; Male ; Treatment Outcome ; Urethra ; surgery ; Urologic Surgical Procedures ; methods

OBJECTIVETo investigate the role of JWA gene in benzo (a) pyrene [B (a) P] induced DNA damage and repair effects in HeLa cells.

METHODSThe antisense JWA express vector (pEGFP-C1-asJWA) was constructed and stably transfected into HeLa cells. JWA deficient HeLa cells (asJWA-HeLa) was then screened and established. The general characteristics of asJWA-HeLa cells were investigated. DNA damage and repair cell culture model was conducted by treating the cells with 50 micromol/L B (a) P plus S9 for 3 hours and then the cells were maintained further 0, 1, 3, and 24 hours for DNA repairing. The damaged DNA was detected by single cell gel electrophoresis assay (comet assay).

RESULTSJWA deficient HeLa cells (with a 31% of JWA protein expression as compared with the control) were obtained successfully. Compared with the empty vector transfected cells (C1-HeLa) and the untransfected HeLa cells, asJWA-HeLa cells were more sensitive to B (a) P exposure and with a delayed DNA repair process.

CONCLUSIONThe JWA determined might function as a potential effective environmental responsive gene and actively participate the process of B (a) P exposure associated with intracellular signal pathways of DNA damage and repair.

Benzo(a)pyrene ; toxicity ; DNA Damage ; drug effects ; DNA Repair ; DNA-Formamidopyrimidine Glycosylase ; deficiency ; genetics ; Gene Expression ; HeLa Cells ; Heat-Shock Proteins ; deficiency ; genetics ; Humans ; Intracellular Signaling Peptides and Proteins ; deficiency ; genetics

To investigate the technological parameters of the isolation and purification of 10-hydroxycamptothecin and vincoside-lactam from Camptotheca acuminata seed by polyamide. The static arid dynamic adsorption characteristics of 10-hydroxycamptothecin and vincoside-lactam on polyamide were studied, and the contents were determined by HPLC. The optimum parameters for adsorption were as follows: the contents of 10-hydroxycamptothecin and vincoside-lactam in the extracts were 0.189 g x L(-1) and 0.334 g x L(-1), respectively, pH 6, flow rate was 1.0 mL x min(-1), processing volume was 3 BV; for desorption: ethanol-water (60:40), flow rate was 1.0 mL x min(-1), 5 BV as an eluent. After treated with polyamide, the contents of 10-hydroxycamptothecin and vincoside-lactam were 17.52% and 32.87%, respectively, the recovery yields were 66.05% and 75.86%, respectively. Results showed that polyamide revealed a good ability to separate 10-hydroxycamptothecin and vincoside-lactam. Therefore, we concluded that results in this study may provide scientific references for the large-scale production of 10-hydroxycamptothecin and vincoside-lactam extracted from C. acuminata seed.
Camptotheca ; chemistry ; Camptothecin ; analogs & derivatives ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Hydrogen-Ion Concentration ; Nylons ; chemistry ; Seeds ; chemistry

In vitro gene delivery, polyethyleneimine (PEI) has been described as one of the most efficient nonviral vector. Herein the formation mechanism of PEI/DNA complexes is elucidated. The transition phase of "bead-on-string" structure in the formation of complexes was supposed to exist through spectroscopy, electrophoresis and transmission electron microscopy (TEM) technology. The construction of PEI/DNA complexes is related closely to the characteristics of PEI and DNA plasmid. As well as the dominant electrostatic effects, the nonelectrostatic interactions were thought to be partially responsible for the presence of PEI/DNA complexes even in the high ionic strength. The surface charge of complexes particles increased with the N/P ratio, but the absolute value of zeta potential was lower at the N/P ratio of 8 and 12, perhaps attributed to the use of larger DNA plasmid. As a result, the repulsion between particles was decreased and prone to aggregate to the structure like a clustered grape-string in the solution. Interestingly, contrast to the formation behavior of complexes, the PEI/DNA complexes aggregated primarily due to hydrophobic interactions while electrostatic attractions play a little role in the complexes particles aggregation in different concentrations of salt solutions. Comparable transfection efficiency in HepG2 cells was observed for the Lipofectamine 2000 and PEI/DNA complexes at the N/P ratio of 12, and showed that larger or aggregable complexes could transfect the cells in some different mechanisms.
DNA ; genetics ; Drug Carriers ; Gene Transfer Techniques ; Genetic Therapy ; Polyethyleneimine ; chemistry ; Transfection

OBJECTIVETo study the expression and clinical significance of ribosomal S6 kinase-4 (RSK-4) in breast cancer and explore the role of RSK-4 in the genesis and development of breast cancer.

METHODThe expression levels of RSK-4 mRNA and protein were detected in 56 cases of breast cancer and the normal breast tissues, as well as in 20 cases of breast benign lesions, by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry.

RESULTSThe expression rates of RSK-4 mRNA in breast cancer, the normal breast tissues and breast benign lesions were 48.2%, 76.8% and 75.0%, respectively. The expression level of RSK-4 mRNA in breast cancer was significantly lower than those in normal breast tissues and breast benign lesions tissues (P < 0.05). The expression level of RSK-4 significantly correlated with tumor size and clinical stage (P < 0.05).The expression rate of RSK-4 protein was 39.3% in breast cancer tissues, which was significantly lower than that of normal breast tissues (71.4%) and breast benign lesions (75.0%, P < 0.01). The expression level of RSK-4 protein was lower in breast cancer with large tumor, high clinical stage and lymph node metastasis. In 56 cases of breast cancer samples, the consistency rate of RSK-4 mRNA and protein was 73.2%. A significant correlation was found between RSK-4 mRNA and protein (χ² = 10.254, P < 0.05).

CONCLUSIONThe down-regulation of RSK-4 expression in breast caner suggests that it is a breast cancer suppressor gene, and the lack or down-regulation of RSK-4 expression is involved in the genesis and progression of breast cancer.

Adult ; Aged ; Breast ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Down-Regulation ; Female ; Fibroadenoma ; metabolism ; pathology ; Humans ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Ribosomal Protein S6 Kinases, 90-kDa ; genetics ; metabolism ; Tumor Burden ; Young Adult