This paper was aimed to study the genetic diversity and genetic relationship of germplasm resource of Lonicerae Japonicae Flos in order to provide references for its breeding. A total of 36 samples from 18 farm varieties and wild species, as well as related species of Lonicera japonica Thunb. from the main production areas were studied by ISSR-PCR markers. The Jaccard coefficient was worked out by NTSYS-pc software. And a cluster dendrogram of different samples was established based on unweighted pair-group method with arithmetic mean (UPGMA). The re-sults showed that 12 ISSR primers generated 129 loci of which 114 loci were polymorphic. The average percentage of polymorphic bands (PPB) is 88.37%. In the cluster dendrogram, different samples of Lonicera japonica are in the same group, which showed that it is a natural species; the wild sample is separated from the cultivated samples; the traditional type-Maohua is relative stable, but the type of Jizhuahua includes complicated varieties, and it has obvi-ous genetic variation; the new variety Jiufeng 1 has already distinct into one stable type. It was concluded that the ISSR method was suitable for germplasm identification, genetic diversity analysis of Lonicerae Japonicae Flos, thus providing a theoretical foundation for its cultivation and breeding.

OBJECTIVETo evaluate the feasibility and efficacy of unilateral pedicle screw fixation in treating thoracolumbar fractures through paraspinal approach.

METHODSFrom January 2006 to January 2009,21 patients with single level thoracolumbar fracture without neurological symptoms were treated with unilateral pedicle screw fixation through paraspinal approach. There were 14 males and 7 females,aged from 21 to 65 years old with a mean of 36.4 years. The duration from injury to operation ranged from 6 h to 5 d with an average of 3 d. According to the classification of Denis fracture, compression fractures happedned in 12 cases and burst fractures happened in 9 cases,including 1 case with T5 fracture, 2 cases with T7 fracture, 2 cases with T10 fracture, 3 cases with T11 fracture, 8 cases with T12 fracture, and 5 cases with L1 fracture. Based on the Flankel grade, all patients were classified as grade E. Anterior vertebral body height ratio, sagittal Cobb angle, condition of internal fixation failure, visual analogue score (VAS) were evaluated.

RESULTSAll patients were followed up from 12 to 36 months with an average of 20.5 months. No internal fixation failure was found. Anterior vertebral body height ratios at preoperative 3 days after operation and last follow-up were 54.3 +/- 2.8, 92.9 +/- 1.5, 93.8 +/- 1.7, respectively;sagittal Cobb angle at the three timepoints were (27.8 +/- 2.5) degrees, (5.3 +/- 0.8) degrees, (6.3 +/- 1.4) degrees, respectively; the difference was statistical significant (P < 0.05). VAS was (1.2 +/- 0.4) points at last follow-up and had obviously improved (P < 0.05).

CONCLUSIONTreatment of thoracolumbar fractures with unilateral pedicle screw fixation through paraspinal approach is safe with the advantages of micro-trauma and less blood loss,which can not only completely retain the posterior spinal complex structure, reinforce the spinal stability, raise the reductional quality, but also improve the strength of fixation and the distribution of stress force.

Adult ; Bone Screws ; Feasibility Studies ; Female ; Fracture Fixation, Internal ; instrumentation ; methods ; Humans ; Male ; Middle Aged ; Spinal Fractures ; diagnostic imaging ; surgery ; Thoracic Vertebrae ; diagnostic imaging ; injuries ; surgery ; Tomography, X-Ray Computed ; Young Adult

AIM: To evaluate the effects of 23G vs 20G pars plana vitrectomy ( PPV ) combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation for macular epiretinal membrane with cataract. ·METHODS: Totally 45 eyes of 45 patients with macular epiretinal membrane and cataract were enrolled in this retrospective non-randomized controlled clinical study. All eyes were treated with PPV combined with internal limiting membrane peeling, phacoemulsification and intraocular lens implantation. There were 20 eyes in 23G PPV group, and 25 eyes in 20G PPV group. The best corrected visual acuity ( BCVA ) , intraocular pressure (IOP), counting of corneal endothelial cells ( CEC) and central retinal thickness ( CRT ) were examined before surgery. BCVA results were converted to the logarithm of the minimum angle of resolution ( LogMAR ) visual acuity. All operations were performed by the same doctor. Operation time for vitrectomy and membrane peeling, average ultrasound energy ( AVE) and effective phacoemulsification time ( EPT ) were recorded. BCVA and CRT were observed postoperatively at 30d and 90d, counting of CEC was observed postoperatively at 90d. IOP was observed postoperatively at 1d and 7d. ·RESULTS:The mean operation time for vitrectomy were 12. 57± 1. 35min in 23G group and 17. 30 ± 1. 19min in 20G group. The difference was statistically significant ( t =-12. 488, P<0. 01). There were no statistical significances in operation time for membrane peeling, AVE and EPT between 23G and 20G groups ( t=-0. 68,-1. 186,-0. 737, P=0. 500, 0. 242,0. 465). On 1d after surgery, IOP in 23G group was lower than that in 20G group, the difference was statistically significant (t= -2. 345, P=0. 024). The BCVA and CRT of the two groups both improved after operations. There were no statistically significant differences between two groups in terms of IOP, BCVA, and CRT ( F = 0. 465, 1. 895, 0. 689; P = 0. 499, 0. 176, 0. 411). IOP, BCVA and CRT were significant statistical different in different time-point within each group ( F=291. 245, 103. 06, 665. 402, P<0. 01 ). Different surgical methods of 23G and 20G had interactive effects on IOP with different time points ( F = 13. 245, P<0. 01 ), but different surgeries had no interactive effects on BCVA and CRT with different time points (F=1. 212, 2. 293;P=0. 283, 0. 129). The counting CEC in 23G group was more than that in 20G group postoperatively at 90d, the difference was statistically significant (t=2. 049, P=0. 048). ·CONCLUSION: The 23G PPV combined with internal limiting membrane peeling, phacoemulsification, intraocular lens implantation for macular epiretinal membrane with cataract is effective. Compared with 20G PPV, 23G PPV has advantages in operation time for vitrectomy and counting CEC. But lower IOP is likely in 23G PPV on 1d after surgery

Objective To exclude false positivities in detection of IgM antibodies against hepatitis E vires of genotype 4 (HEV-4) using a truncated immunodominant polypeptide of HEV open reading frames (ORF2). Methods The recombinant ORF2 immunodominant polypeptide corresponding to amino acids (AA) 459-607 and a truncated polypeptide corresponding to AA 472-607 were separately applied to coat ELISA plates. Anti-HEV IgM from 35 serum samples with HEV RNA positive, 69 serum samples from healthy individuals and 117 clinically suspicious HEV RNA positive serum samples was detected by an indirect ELISA and was confirmed by western blot in protein level and RT-PCR detecting in RNA level. Results Western blot analysis showed that the sera from HEV patients reacted with the dimmer of peptide 459-607, but they didn't react with the monomer and peptide 472-607. The ELISA showed that all 35 serum HEV RNA positive samples reacted with peptide 459-607 but not with peptide 472-607 and none of the 69 serum samples from healthy individuals reacted with either polypeptide. Among 117 chnically suspicious HEV RNA serum samples, 5 samples reacted simultaneously with both polypeptides. But the difference between 450 nm absorbance (A450) value was less than 0. 5. Western blot analysis demonstrated that all the 5 serum samples were anti-HEV IgM- negative. The 5 serum samples was detected negative by RT-PCR, indicating that the false pesitivities were caused by non-specific absorption. Conclusions ORF2 peptide 459-607 may be used to detect anti-HEV lgM efficiently. The false positivities caused by non-specific absorption can be largely excluded according to the difference between 45Ohm absorbance (A450) value when serum reacts with both polypeptides.

Background Antagonists against vascular endothelial growth factor (VECF) play key roles in treating and preventing neovascular ophthalmopathy. As a novel anti-angiogenic factor, insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) might be an antagonist against VEGF in eye. Objective This study was to explore the inhibitory effect of IGFBP-rP1, a novel anti-angiogenic factor, on VEGF-induced retinal angiogenesis in vitro. Methods The retina-choroid endothelial cell line ( RF/6A ) was cultured in DMEM containing 10% fetal bovine serum. Culture cells were divided into control group(free-serum culture group) ,10mg/L VEGF culture group and different concentrations of IGFBP-rP1 (50,100,200 mg/L) +10 mg/L VEGF group. The expression of IGFBP-rP1 in the cells was detected by immunofluorescence assay. The proliferation and migration of RF/6A cells were evaluated using MTS colorimetric assay and the chemotactic motility assay, respectively. Flow cytometry was used to detect the apoptosis of RF/6A cells. Results The immunofluorescence assay RF/6A cells showed the green fluorescence in cytoplasm and red fluorescence in nuclei after cells were exposed to any concentration of IGFBP-rP1 ,but only red fluorescence was seen in nuclei in control cells. After stimulation of 10 mg/L VEGF,the proliferation value (A490) was elevated and the numbers of cell migration were increased in comparison with control group (t = -15. 191, P = 0. 000; t = -21. 274, P = 0. 000 ) , but the cellular apoptosis rate was lower than the control group (t - 10. 228, P = 0. 000 ) . After treated with various concentrations of IGFBP-rP +10% VEGF, the proliferation and migration of RF/6A cells were significantly decreased in comparison with only 10% VEGF group (F = 534. 158,P = 0. 000;F = 2742. 323,P = 0.000,respectively) ,and the inhibitory effects were gradually enhanced with the increase of IGFBP-rP1 levels (P<0. 05). The apoptosis rate of RF/6A cells in 50,100 and 200 mg/L + 10 mg/L VEGF groups increased by ( 1. 26±0. 04)% ,( 1. 50±0. 07)% and ( 1. 93±0. 27)% respectively,showing significant differences among different groups ( F = 274. 273, P = 0. 000). Conclusion IGFBP-rP1 inhibits the proliferation and activity of retina and choroid endothelial cells induced by VEGF at a concentration-independent manner. It appears to be as a novel endogenous inhibitory factor in retinal angiogenesis.

Adult ; Carcinoma, Renal Cell ; secondary ; Female ; Humans ; Kidney Neoplasms ; pathology ; Magnetic Resonance Imaging ; Vaginal Neoplasms ; secondary

OBJECTIVETo investigate the molecular epidemiological characteristics and antibiotic resistance profiles of Vibrio cholerae O139 in Shandong province.

METHODSA total of 13 strains of V. cholerae O139 (9 clinical strains and 4 environmental strains) isolated from cholera epidemics in Shandong province since 1997 were recovered and confirmed with serum agglutination and biochemical reaction. Pulsed-field gel electrophoresis (PFGE) was carried out for molecular subtyping. Virulence genes and drug resistance related genes were detected by PCR. Antibiotic susceptibility tests were performed using micro-broth dilution method.

RESULTSThirteen strains of V. cholerae O139 were differentiated into seven pulsetypes. One clinical strain and two environmental strains isolated from Jining in 2013 were clustered into the pulsetype namely KZGN11O139. CN0077, and an identical PFGE pattern of KZGN11O139. CN0002 was found among three clinical strains from Jinan in 2005, Jining in 2005 and Heze in 2009. Other pulsotypes were unique in China and found only in Shandong province. Because of deletion of ctxAB and tcpI, the PFGE patterns of two strains isolated from Yantai in 2000 and 2004 were different from other 11 strains which harbored ctxAB, tcpA, tcpI, rtxA, hlyA and toxR. All strains contained one or more drug resistance related genes such as intI 1, intI 4 and sxt, and were resistant to two kinds of antibiotics at least. Among the 12 kinds of antibiotics, the resistant ratioes to kamamycin, trimethoprim-sulfamethoxazole, ampicillin and gentamicin were 11/13, 9/13, 7/13 and 7/13, respectively.

CONCLUSIONMolecular subtyping indicates possible epidemiological links among V.cholerae O139 in Shandong province, and almost all strains were toxigenic and drug resistant.

China ; Cholera ; Cholera Toxin ; Drug Resistance, Bacterial ; Electrophoresis, Gel, Pulsed-Field ; Epidemics ; Humans ; Molecular Epidemiology ; Polymerase Chain Reaction ; Vibrio cholerae O139 ; Virulence

Regulation of the activity of CYP450 has always been research focus of drug metabolism. The effect of compounds on the mRNA and protein expression level of CYP450 is the main purpose of most of the existing reports. In recent years, the protein modification in the posttranslation level has been found to participate in maintaining the proper function of CYP450, thus effect of posttranslational modification on the enzyme activity has been paid more and more attention. Posttranslational modifications including phosphorylation, nitration, and ubiquitination have been described to regulate the activity of CYP450. In this paper, recent developments in the effects of posttranslational modifications on the activity of CYP450 have been reviewed.
Animals ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Glycosylation ; Humans ; Nitric Oxide ; metabolism ; Oxidation-Reduction ; Phosphorylation ; Protein Processing, Post-Translational ; RNA, Messenger ; metabolism ; Ubiquitination

An HPLC method for the determination of 18alpha-glycyrrhetinic acid and 18beta-glycyrrhetinic acid in rat plasma was established, which was used subsequently to determine the pharmacokinetic profiles of both epimers of glycyrrhetinic acid in rats. alpha-glycyrrhetinic acid, beta-glycyrrhetinic acid, and a mixture of alpha-glycyrrhetinic and beta-glycyrrhetinic acids were administered to rats via gastric infusion. Blood samples were collected at different time intervals and extracted by liquid-liquid extraction. Separation was achieved by using a Kromasil C18 column (150 mm x 4.6 mm, 5 microm) with the mobile phase composed of acetonitrile--4 mmol x L(-1) ammonium acetate solution (46 : 54, v/v) at a flow rate of 1.0 mL x min(-1), and the detection wavelength was set at 250 nm. The pharmacokinetic parameters were calculated using the software DAS 2.0. In a combined administration, the main pharmacokinetic parameters of beta-glycyrrhetinic acid are significantly different from that of alpha-glycyrrhetinic acid (P < 0.05), while no significant difference was obtained when administrated individually. Compared to the single administration, significant differences (P < 0.05) on the values of AUC(0-t) and AUC(0-infinity) of beta-glycyrrhetinic acid were observed when this chemical was administrated together with alpha-glycyrrhetinic acid. In contrast, the pharmacokinetic parameters of alpha-glycyrrhetinic acid were not affected even under the co-administration. Here, a sensitive, specific, rapid and reproducible HPLC method was developed for the pharmacokinetic studies of alpha-glycyrrhetinic acid and beta-glycyrrhetinic acid in rat plasma.
Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Glycyrrhetinic Acid ; analogs & derivatives ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Stereoisomerism

AIMTo investigate the changes in the expression of four kinds of calcium regulatory proteins mRNA on the isolated ischemia/ reperfusion (IR) hearts.

METHODSThe rat hearts were divided into two groups: control group and IR group which received 45 min exposure to Krebs-Henseleit solution after 15 min zero-flow global ischemia. The indexes of left ventricular function, such as LVDP, +dp/dt(max), -dp/dt(max), and an arrhythmia scoring system were compared between the two groups. The messenger ribonucleic acid (mRNA) amount of sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA), phospholamban (PLB), inositol 1,4,5-trisphosphate receptor2 (IP3R2) and ryanodine receptor2 (RyR2) was measured by reverse transcription-polymerase chain reaction (RT-PCR) and normalized to the mRNA levels of beta-actin.

RESULTSIn the IR group, LVDP, +dp/dt(max) and -dp/dt(min) of the isolated hearts were depressed and the high rate of arrhythmias occurred during reperfusion. The levels of SERCA, IP3R2, RyR2 mRNA were lower in the IR isolated hearts group than those in the control group, while there was no difference in the level of phospholamban.

CONCLUSIONThese data suggest that myocardial ischemia/reperfusion can induce the depression of cardiac performance and an increased risk of arrhythmias, concomitant with the decrease in SERCA, IP3R2, RyR2 mRNA steady state levels.

Animals ; Myocardial Reperfusion Injury ; metabolism ; Myocardium ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; genetics ; metabolism