Female ; Follow-Up Studies ; Hearing Loss ; etiology ; Hearing Tests ; Humans ; Infant, Newborn ; Male ; Risk Factors

This paper summarize the practice in the teaching reform of microbiology experiment in recent years. We identify the main contents of experimental teaching systems and pay much more attention to peo-ple-oriented. Through the reform of teaching and assessment methods,students are trained to cultivate their practical ability and spirit of innovation.

A new flavonoid glycoside, (-)-2S-8-methyl-5,7,4'-trihydroxyflavanone-7-O-β-D-glucopyranoside (1), along with five known ones, quercetin-3-O-(2"-galloyl)-α-L-arabinoside (2), kaempferol-3-O-α-L-arabinoside (3), guaijaverin (4), trifolin (5) and hyperin (6), was isolated from the leaves of Eucalyptus robusta. Their structures with absolute configurations were elucidated by NMR, HR-ESI-MS, CD spectra data and physicochemical methods. In addition, 2-6 were isolated from E. robusta for the first time.
Eucalyptus ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Plant Leaves ; chemistry

Objective To explore effect of endogeneous gangliosides(Gls) and integrin ?2?1 on protein phosphotyrosine expression of pp125 focal adhesion kinase (pp125FAK) after adhesion of SK-N-SH neuroblastoma cells to collagen(Col).Methods SK-N-SH cell line with high expression of integrin ?2?1 was cultured in presence of D-threo-1-phenyl-2-decanolamino-3-morphinoline-1-propanol(D-PDMP).Effect of endogeneous Gls,anti-?2 and anti-?1 monoclonal antibody on protein phosphotyrosine expression of pp125FAK during adhesion of SK-N-SH cells to Col were determined by immunoprecipitate and Western blotting.Results After 6 days,endogenous Gls in cells were almost depleted.Gls-depletion,anti?2 and anti-?1 monoclonal antibody were able to decrease pp125FAK expression of SK-N-SH cells adherent to Col respectively.GD2,the major component of neuroblastoma cell Gls could reco-ver pp125FAK expression to a certain degree.Conclusions Endogenous tumor Gls regulate protein phosphotyrosine expression of pp125FAK during adhesion of neuroblastoma cells to Col.It is suggested that tumor Gls may increase signal transduction of tumor cell integrin ?2?1 by increasing tyrosine phosphorylation of pp125FAK.

Objective To assess the role of pelvic ultrasound examination in discriminating between normal girls,isolated premature thelarche/pubarche /menarche and central precocious puberty (CPP).Methods Eighty-four isolated premature thelarche/pubarche /menarche cases,47 CPP cases,and 177 normal girls aged 0-10 years were recruited.All diagnoses were confirmed by the gonadotropin-releasing hormone-stimulation test.All subjects underwent pelvic ultrasound examination for the measurement of length,width,thickness,and volume of the uterine body,uterine cervix,and ovary,and the number of follicles with diameter≥ 4 mm.The groups were subdivided by age intervals when the difference in ultrasound measurements between CPP,isolated premature thelarche/pubarche/menarche,and normal girls were analyzed.Results 1) Differentiation between CPP and normal girls:for the 6-8 years,there were 11 variables elevated in CPP as compared to the normal girls.Uterine cervix thickness was the most efficient parameter as judged by the largest value of area under the ROC curve (0.958).The best cut-off,sensitivity,and specificity was 0.73 cm,93.30％,and 85.70％ respectively;for the 8-10 years,uterine body volume was the best parameter among the 10 elevated variables as judged by the largest area under the ROC curve (0.869),3.23 cm3 was the best cut-off limit with a sensitivity of 84.21％ and a specificity of 52.11％.2) Differentiation between isolated premature thelarche/pubarche/menarche and normal girls:for the 0-6 years,ovary thickness was the best variable as judged by the largest area under the ROC curve (0.806),0.98cm was the best cut-off limit with a sensitivity of 76.46％ and a specificity of 84.85 ％ ;for the 6-8 years,ovary width was the best variable among the 8 valuable variables for its largest area under the ROC curve (0.843),1.39 cm was the best cut-off limit with a sensitivity of 85.71％ and a specificity of 73.81％ respectively;for the 8-10 years,uterine cervix thickness was the best variable among the 5 valuable variables for its largest area under ROC curve (0.841),0.75 cm was the best cut-off limit with a sensitivity of 90.48％ and a specificity of 64.21％.3) Differentiation between CPP and isolated premature thelarche/ pubarche/menarche cases:for the 6-8 years,uterine cervix length and width were potential parameters.Uterine cervix length was the best variable for its largest area under the ROC curve(0.764),and 1.49 cm was the best cut-off limit,the corresponding sensitivity and specificity was 93.33％ and 55.17％ respectively;for the 8-10 years,3 variables could be used,among which uterine cervix length was the best variable for its largest area under the ROC curve (0.893),1.88 cm was the best cut-off limit with a sensitivity of 100％ and a specificity of 71.43％.Condusions Pelvic ultrasound examination is a valuable tool for the diagnosis and differentiation between CPP,isolated premature thelarche/pubarche /menarche and normal girls.

BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

ObjectiveTo investigate the role of 15-F2t-isoprostane in intestinal injury induced by intestinal ischemia/reperfusion (I/R) in rats.MethodsThirty-two pathogen free adult male SD rats weighing 230-255 g were randomly divided into 4 groups ( n =8 each):group sham operation (group S) ; group intestinal I/R; group SQ-29548 (TXA2 receptor antagonist) (group SQ) and group DMSO (the solvent).Intestinal I/R was induced by 60 min occlusion of superior mesenteric artery (SMA) followed by 120 main reperfusion in groups I/R,SQ and DMSO SQ-29548 2 μmol/kg and DMSO were injected subcutaneusly at abdominal wall at 30 min before SMS in groups SQ and DMSO respectively.Arterial blood samples were taken at 120 min of reperfusion for determination of serum diamine oxidase (DAO) activity and 15-F2t-isoprostane,endothelin-1 (ET-1) and thromboxane B2 (TXB2) concentrations.Intestinal tissues were removed for microscopic examination and determination of myeloperoxidase (MPO) and SOD activities,MDA and lactate contents.Intestinal damage was assessed and scored according to Chiu (0 =normal,5 =disruption of tunica propria,bleeding and ulceration).ResultsIntestinal I/R significantly increased Chiu's score,MDA and lactate contents and MPO activity and decreased SOD activity in intestine in group I/R as compared with group S.SQ-29548 pretreatment significantly decreased Chiu's score,lactate content and MPO activity in intestine and increased intestinal SOD activity and decreased serum DAO activity and ET-1 concentration in group SQ as compared with group I/R.Conclusion15-F2t-isoprostane is involved in the development of intestinal injury induced by intestinal I/R by activating TXA2 receptor,increasing ET-1 production and promoting neutrophil infiltration.

Background Platelet-derived growth facto(PDGF) affectthe proliferation of human lenepithelial cell(LECs),and human LECexpresPDGF-α recepto(PDGFR-α) throughoutheilifetime.The binding of activated PDGF-α receptowith PDGF promotethe synthesiof DNA.Othestudiedemonstrated thasilencing of PDGFR-α by antisense oligodeoxynucleotide(ASODN) inhibitthe growth of RPE cellin proliferative vitreoretinopathy (PVR),buwhethethitechnique ifeasible foLECiunclear.Objective Thistudy wato investigate the effecof the knockdown of the PDGFR-α on the proliferation of human LECin vitro,and to offean experimental basifothe gene therapy of posteriocapsule opacification.MethodHuman LECstrain SRA01/ 04 wacultured in α-MEM containing fetal bovine serum.The cellwere incubated in 6-well platea5 × 104 cells/ well and transfection of ASODN-containing liposome waperformed.The cellwere divided into the blank control group (with blank liposome),PDGFR-α missense oligodeoxynucleotide(MSODN) group (with PDGFR-α MSODN + liposome),0.5 μmol/L PDGFR-α ASODN group (with 0.5 μmol/L PDGFR-α ASODN+liposome) and 1.0 μmol/L PDGFR-α ASODN group (with 1.0 μ mol/L PDGFR-α ASODN+liposome).The morphology of LECwaexamined undean inverse microscope 24 houraftetransfection.The expression of PDGFR-α mRNin the cellwadetected by reverse transcription-PC(RT-PCR).The rate of proliferation (A490) of the cellwaassayed using Mtand the inhibitory rate of PDGFR-α ASODN on proliferation wameasured.The percentage of LECin G1 phase waanalyzed by flow cytometer.ResultThe LECgrew well and exhibited polygonal shape in the blank control group and PDGFR-α MSODN group 24 houraftetransfection.Buin the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups,the cellappeared round in shape and the numberof cellwere obviously decreased.The expression of PDGFR-α mRNdetected by RT-Pcdemonstrated highelevel in the blank control group and PDGFR-α MSODN group;however,the PDGFR-α mRNexpression waobviously lowein the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups.The A490 value wa0.661 ± 0.036,0.655 ± 0.016,0.529 ± 0.030 and 0.441 ± 0.039 in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,respectively,showing significandecline in the 0.5 μmol/L PDGFR-α ASODN group and 1.0 μ mol/L PDGFR-α ASODN group in comparison with the blank control group (F=34.08,P＜0.01).The percentageof LECin G1 phase were (47.73±1.18)％,(49.48±1.09)％,(53.31±1.30)％ and (59.98±0.95) ％ in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,showing significandifference among them (F =68.41,P＜0.01),and thain the 0.5 μmol/L PDGFR-α ASODN group o1.0 μmol/L PDGFR-α ASODN group showed significantly increase in comparison with the blank control group (P＜0.05).ConclusionPDGFR-α silencing could inhibithe proliferation of human LECin vitro.

An endophytic bacterium strain EBS05 from Cinamonum camphra was identified as Bacillus subtilis by morphological taxonomy and sequence analysis of 16S~23S rRNA intergenic spacer regions. Properties of antimicrobial compound produced by EBS05 were assayed. The active compound had the maximum absorbance peak at ?213.5 nm. The antimicrobial activity was stable in solution with pH value from 5 to 8, and decreased significantly in solution with pH value less than 4.0 or more than 9.0. The antimicrobial compound had thermodynamics stability. Its activity changed a little after treated at 60?C~80?C for two hours, and compared with 65% original activity after treated at 1?105 Pa for 30 minutes. The active substance had high resistance to ultraviolet radiation and protease K. Antimicrobial compound was soluble in alcohol solu- tion, which was easily dissolved in methanol and ethanol, but not dissolved in ethyl acetate, acetonitrile and petroleum et al.

OBJECTIVETo investigate the effects of the attachment site of grafts and the tunnel angle on the function of knee joint after anterior cruciate ligament reconstruction.

METHODSFrom January 2006 to May 2009, 47 patients(32 males and 15 females, ranging in age from 19 to 51 years old, with an average of 35.3 years old) were treated with single-bundle reconstruction of anterior cruciate ligament. Several indexes were measured at the latest follow-up as follow: attachment sites of graft on the femoral condyle were recorded, the femoral tunnel angles on coronal and sagittal planes were measured on postoperative X-ray films. According to the IKDC score, these patients were divided into two groups. In the first group, 38 patients were found the IKDC score more than 90 at the latest follow-up, and in the second group 9 patients were found IKDC score less than 90. By comparing the two groups, the relation of the indexes and postoperative function of knee was analyzed.

RESULTSThe IKDC which was more than 90 at the latest follow-up was found in 38 patients, whose femoral attachments site of ACL was positioned at (29.73 +/- 4.31)% (ranged from 16.21% to 53.82%) from the posterior end of Blumensaat's line. IKDC which was less than 90 was found in 9 patients, whose femoral attachments site of ACL was positioned at (46.61 +/- 3.43)% (ranged from 27.18% to 72.34%). There was significant difference between the two groups (P = -0.000 7). The IKDC more than 90 at the latest follow-up was found in 38 patients,whose femoral tunnel angle on coronal plane was (49.5 +/- 4.72) degrees (ranged from 33 degrees to 67 degrees) and on sagittal plane was (31.3 +/- 5.12) degrees (ranged from 11 degrees to 45 degrees) were significantly less than those whose IKDC less than 90 at the latest follow-up on coronal plane was (67.6 +/- 3.09) degrees (ranged from 41 degrees to 81 degrees) and on sagittal plane was (41.2 +/- 5.69) degrees (ranged from 23 degrees to 56 degrees) (P = 0.000 7, P = -0.000 8).

CONCLUSIONThere is close relation between the attachment site of grafts and the tunnel angle with the function of knee, so in anterior cruciate ligament should be anatomic reconstructed with the anterior medial approach.

Adult ; Anterior Cruciate Ligament Reconstruction ; Female ; Femur ; surgery ; Humans ; Knee Joint ; physiopathology ; Male ; Middle Aged