By exploring the different components of the lysis buffer and optimize the 2-DE conditions,established the best proteomics technical system for Populus euphratica's heteromorphic leaves,while take the heteromorphic leaves in the same blanche as the test materials to find differences between the protein expressions of the leaves.It showed that the lysis solution which containing 2mmol/L thiourea,7mmol/L urea,2% CHAPS,60mmol/L DTT and 0.2% IPG buffer could dissolve the protein better.Through tandem mass spectrum,the results show that heteromorphic leaves are different in photosynthesis and respiration.This research offered valuable informations for understanding the molecular mechanism during leaves development and elucidating the mechanism of the eco-adaptability of Populus euphratica.

Objective To detect the serum antibody to HPV6 in patients with condylomata acuminata (CA) by use of human papilloma virus(HPV)6bL 1 virus-like particles (VLPS), and to assess the significance of detecting HPV infection. Methods HPV6bL 1 VLPS were produced by a HPV6bL 1 recombinant baculovirus in insect cells,purified by CsCl density gradients centrifugation,and used for ELISA to screen 136 human serum samples in three groups. Results The prevalence of antibody to HPV6bL 1 VLPs was much higher in patients with codylomata acuminata (75%Ab+;mean abs 0.111?0.094) than that in patients with cervical cancer (14.3%Ab+;mean Abs 0.029?0.022)and that in control groups (2.9%Ab+;mean Abs 0.012?0.024). The differences among three groups are significant(P

Adult ; Female ; Humans ; Kidney ; pathology ; Kidney Neoplasms ; pathology

Objective To investigate the effect of the polymorphisms of multidrug resistance 1 (MDR1) C3435T and G2677T on Tacrolimus (Tac) individualized treatment and prognosis of grafts in the renal transplantation recipients (RTRs).Methods One hundred and twenty-seven RTRs who treated with Tac regimen and had a stable graft function were enrolled,and were divided into adjuvant treatment group and non-adjuvant treatment group according to whether given adjuvant drugs to raise Tac trough concentrations. MDR1 C3435T and G2677T SNPs were detected by using sequence specific primers PCR.Tac trough concentrations of whole blood were measured by using enzymelabeled immunosorbent assay.Tac concentration-to-dose ratio (C/D) standardized by body weight was compared according to the various genotypes and haplotypes of MDR1 C3435T and G2677TA SNPs.Results Adjuvant treatment group including 36 recipients had a higher frequency of C genotype of C3435T than un-adjuvant treatment group (68.05％ vs 48.35％,P ＜ 0.01 ). The frequency of G2677TA polymorphisms was of no significant difference between the two group recipients (P＞ 0.05).As to non-adjuvant treatment recipients,the mean Tac DD required and C/D were not significantly different among various polymorphisms of MDR1 G2677T/A and C3435T or various haplotypes (P＞0.05).During A follow-up period of 4 years,13 recipients suffered graft dysfunction in which 84.6％ (11/13) carried 3435C genotype (P＞0.05).Conclusion The frequency of MDR1 C3435T polymorphisms in RTRs is high in the recipients given adjuvant treatment to raise Tac concentrations.Recipients with 3435C genotype were prone to graft dysfunction.

OBJECTIVETo explore the clinical effect of the sacrococcygeal space injection for the treatment of failed back surgery syndrome.

METHODSFrom July 1998 to October 2012,47 patients with failed back surgery syndrome were treated and included 39 males and 8 females with an average age of 61.5 years old ranging from 35 to 89 years old. Among them,41 patients experienced one time of operation, 6 patients with twice of operation. Forty-one patients underwent single,bilateral fenestration or central laminectomy decompression, discectomy. Six patients underwent total laminectomy discectomy and inter body fusion and pedicle screw fixation. All patients were examined by X-ray plain film, CT or MRI before treatment. The anticoagulation was discontinuation before treatment. The needle was put into the sacrococcygeal gap at prone position in the sense of frustration,suction without cerebrospinal fluid and blood,with injection of Mailuoning (Chinese characters: see text) 15 ml. The pain was assessed by VAS before and after treatment. The Oswestry low back pain disability index and survival quality interference degree were evaluated.

RESULTSAt 1 month after treatment,the pain VAS decreased from 59.24 +/- 17.35 before treatment to 19.19 +/- 11.19 after treatment (P < 0.05); The Oswestry low back pain disability index decreased from (41.35 +/- 9.87)% before treatment to (23.17 +/- 17.56)% after treatment (P < 0.05); The survival quality interference degree decreased from 6.5 +/- 2.2 before treatment to 2.6 +/- 1.4 after treatment (P < 0.05).

CONCLUSIONThe sacrococcygeal gap injection for treatment of failed back surgery syndrome has advantages of simple, safe, fewer complications, and low treatment cost.

Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; administration & dosage ; Failed Back Surgery Syndrome ; diagnostic imaging ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Radiography ; Sacrococcygeal Region ; diagnostic imaging

OBJECTIVETo observe the intervention of Shenkangling Decoction (SD) on the renal injury of primary nephrotic syndrome (PNS) children patients of Shen deficiency blood stasis syndrome (SDBSS) and to explore its mechanism.

METHODSTotally 65 PNS children patients were randomly assigned to the combined group (33 cases, treated by SD +Western medicine) and the Western medicine group (32 cases, treated by Western medicine). Meanwhile, 30 healthy children were recruited as the healthy control group from the medical examination center. Those in the Western medicine group were treated with prednisone (5 mg per tablet) at the daily dose of 1.5 -2.0 mg/kg till two weeks after their urine protein turned to negative. Then the dosage was reduced once daily per every other day. The therapeutic course lasted for more than 1 year. For those with no effect of prednisone or partial effect, cyclophosphamide intravenous pulse therapy was additionally applied for 2 successive days per week, a total of 6 times, or they took cyclosporine A. Patients in the combined group additionally took SD while starting treatment of prednisone. SD was decocted in water for oral dose, once daily, taken in two portions until 2 months after prednisone was discontinued. Efficacy was evaluated based on serum levels of chemotactic factor CXCL16, disintegrin metalloproteinase 10 ( ADAM10 ), disintegrin metalloproteinase 17 (ADAM17), albumin (ALB), total cholesterol (TC), and 24-h urine protein excretion (UPE) detected by ELISA before and after treatment.

RESULTSCompared with before treatment in the same group, levels of CXCL16, ADAM10, ADAM17, TC, and 24-h UPE were significantly lower in the two treatment groups (P <0. 01). Compared with the control group, levels of CXCL16, ADAM10, ADAM17, TC, and 24-h UPE significantly increased, and the serum ALB level decreased in the two treatment groups (P <0.01). Compared with the Western medicine group at the same time point, levels of CXCL16, ADAM10, ADAM17, TC, and 24-h UPE significantly decreased in the combined group. The 1 -year recurrence rate and the recurrence times decreased in the combined group (P <0.01). The complete remission rate increased in the combined group (P <0.01).

CONCLUSIONSD could effectively improve the clinical prognosis of PNS children patients possibly by reducing the release of inflammatory mediators such as CXCL16, ADAM10, and ADAM17, decreasing UPE and the TC level, and elevating the serum ALB level.

Child ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Nephrotic Syndrome ; drug therapy ; Prednisone ; Syndrome

Objective To investigate the protective effect of hydrodynamics-based injection with plasmids of IL-10, TGF-β1 and TGF-β1 + IL-10 in murine skin transplantation model. Methods Plasmids were constructed by inserting coding sequences of IL-10 and TGF-β1. In F1 mice (Balb/c×C57BL/6, H-2b/d) to Balb/c (H-2d) murine skin transplant model, 20 μg plasmid (blank or IL-10 or TGF-β1 or IL-10 + TGF-β1) was injected to donors by hydrodynamics-based method in first day and every interval 2 days for 6 times. The survival of grafts was observed after 7 days of transplantation. After C56BL/6 spleen cells transfused Balb/c accepted 5 times hydrodynamics-based injection as above,CD4+ CD25+ T regulatory cells of spleen were measured by FACS. Results The survival time of graft in each group was (13.50±1.04)days (blank group), (13.83±1.16)days (IL-10 group), (15.33±1.50) days (TGF-β1 group), and (21.33±3.20) days (IL-10 + TGF-β1 group),respectively (P<0.05). The percentage of CD4+ CD25+ cells was (6. 58±1.86)% (blank group),(10.52±1.13)% (IL-10 group),(14.44±0.42)% (TGF-β1 group),and (14.25±1.24)% (IL-10+TGF-β1 group) respectively (P<0.05). Conclusion Hydrodynamics-based transfection of IL-10 combined with TGF-β1 can synergistically enhance the percentage of CD4+ CD25+ T cells and prolong the graft survival.

Objective To establish a universal stem loop primer (USLP) based real-time PCR method to scan mature miR profile and quantify it's expression.Methods The common universal stem-loop primer pairs were re-designed; 8 random nucleotides were introduced at 3 ' end for reverse transcription of the mature miR,establishing a miR scanning and quantifying system based on SYBR Green Ⅰ PCR (improved USLP method).10-fold gradient diluted standard miRNA-155 cDNA ( 1 ～ 109 copies/μ1) were utilized to evaluate the sensitivity of this method.The specificity was verified by melting curve assay; the precision was assessed by intra-assay coefficient of variation (ICV) of threshold cycle (Ct value) through 20 repeated detections of the standard miR-155 cDNA (2 × 105,2 × 106,2 × 107 copies/μl) ; cost of the primers and time were evaluated,compared with that of the conventional USLP method.Peripheral blood samples were cultured with phytohaemagglutinin (PHA) for0 h,16 h,24 h,48 h and 72 h,and 87 candidate miR that may be associated with human immunity from PubMed data were scanned and quantified from the cultured T cells.Results The sensitivity of the improved USLP method was 103 copies/μl of standard miR-155 cDNA.Melting curve assay showed a single melting peak at 80 ℃,suggesting the excellent PCR specificity of miR-155.Precision of our method quantifying miR-155 was acceptable (ICV ＜ 2.5％ ).Compared with the traditional stem loop primers,our method saved 75％ cost of primers ( 1 917 bp vs 7 851 bp) and 60％ test time of reverse transcription (85 min vs 205 min).By our method,85 of the 87 miR expression in T cells had no significant difference after the PHA stimulation; the expression of miR-150 (72 h) decreased by 10 times and that of miR-155 (48 h) increased 8 times after culture with PHA (Z =-2.032,P =0.042;Z =- 2.023,P =0.043,respectively ).Conclusions The improved USLP method is fast,precise,sensitive,and cost-effective.It could be used for miR profile scanning and quantifying in T cells.

Objective To develop the hypothesis ‘saturated or non-saturated cytotoxicity model' and explain the various phenomena of antibody mediated immunoresponses in recipients,including rejection and accommodation.Methods The imitating complement dependent cytotoxicity.The threshold set to identify as saturated or non-saturated cytotoxicity depends on antigen-antibody complex(R)whether or not above lethal number(D)in effective time.Feasibility of the hypothesis was examined through explaining various phenomena mediated by anti-donor antibodies,especially some contradictory phenomena.Results Hyperacute rejection,accelerated rejection and acute rejection could be well explained by saturated cytotoxicity.Accommodation of ABO imcompatible transplantion,de novo antibody induced injury,change of protein profile,and C4d deposition in graft could be well elucidated by the hypothesis.Conclusion The hypothesis saturated or nonsaturated cytotoxicity model' help to interpret and interconnect various phenomena of antibodies mediated immune response,such as rejection and accommodation.

Objective: To detect the bioavailability of 21 types of perfluorochemicals (PFCs) in rat liver and to examine the effect of protein powder. Methods: Twenty-four rats of the SD strain were randomly divided into the control group, the model group, and the protein powder group. Twenty-one types of PFCs were mixed at an equal concentration of 10 ng/mL, and rats in the model group and the protein powder group were given by oral administration of PFCs mixtures at a daily dose of 5 mL/kg. Rats in the protein powder group were given protein powder by gavage at a dose of 15 mL/kg, while animals in the model and control groups were given deionized water at doses of 15 and 20 mL/kg for 28 successive days. The PFCs contents were quantified in rat liver using ultra-high performance liquid chromatography-electrospray tandem mass spectrometry (UPLC-MS/MS), and the bioavailability was estimated. Results: There were no significant differences in rat body weight or liver/body weight ratio in the control, model and protein powder groups (P>0.05). There were no significant differences in the bioavailability of perfluoroalkylated carboxylic acid (PFCA) or sulfonate (PFSA) in the liver of female and male rats between the protein powder group and the model group (P>0.05), and the gross bioavailability of PFCA (t=-22.266, P<0.001) and PFSA (t=-34.312, P<0.001) was significantly higher in the liver of male rats than in that of female rats in the model group, and the bioavailability of PFCA and PFSA increased followed by a reduction in rat livers with the increase of carbon chain length in the model group. In the model group, the highest bioavailability was measured in perfluorododecanoic acid (PFDoA) and sodium perfluorooctylsulfonate (L-PFOS) in the female rat liver [(36.06±2.93)% and (37.11±1.73)%], and the highest bioavailability was measured in perfluorononanoic acid (PFNA) and L-PFOS in the female rat liver [(61.02±2.16)% and (87.16±3.29)%]. Conclusions The bioavailability of PFCs correlates with the carbon chain length and animal gender in rat livers, and protein powder poses no clear-cut effects on the bioavailability of 21 types of PFCs in rat livers.