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Objective To clne huma PAIl gene and prepare its monoclonal antibodies(McAbs) for determination of its expression in breast cancer cells.Methods Human PAI1 gene eDNA was amplified by RT-PCR from human breast cancer cell line MDA231 and inserted into the prokaryotic expression vector, which expressed fusion protein of MS2-PAI1 in E.coli.Fusion protein of MS2-PAI1 was purified and used for immunizing BALB/C mouse.Traditional hybridoma technology was used to produce hybridoma cells for preparation of monoclonal antibodies.Western blot and immunohistochemistry were used to detect PAI1 expression in breast cancer.Results The 1209 bp full PAI1 gene was cloned.The two hybridoma cell lines that secreted specific monoclonal antibodies against human PAI1 were identified by ELISA.The immunoglobulin subclasses of the McAbs were IgG1.The McAbs can specifically recognize PAI1 but not other proteins.Western blot showed that the McAbs against PAI1 can specifically react with MS2-PAI1 fusion protein and endogenous proteins in cells.The positive reaction was found in breast cancer cell line MDA231 and breast cancer tissues by immunochemical staining.Conclusions The McAbs against human PAI1 are successfully prepared by hybridoma technology with MS2-PAI1 fusion protein expressed in E.coll.It has been shown that PAI1 can be expressed in MDA231 and breast cancer tissues.The McAbs against PAI1 could be a useful tool for the further study of the human PAI1 functions and detection of clinical tumor samples.

Procedure for prolapse and hemorrhoids( PPH) is one of the important techniques developed for the treatment of hemorrhoids with severe degree in the last decade. Its principle is based on the "anal cushion" theory.Compared with traditional hemorrhoidectomy , PPH has advantages of shorter operation time , minor degree of postoperative pain , shorter hospital stay and quicker recovery.However, the occurrence of relapse and re-prolapse of hemorrhoids is high. Besides, the short-term efficacy of PPH for the constipation outlet obstruction caused by anterior rectocele is also favorable.
Anal Canal ; Hemorrhoids ; diagnosis ; Humans ; Operative Time ; Pain, Postoperative ; Prolapse ; Surgical Stapling

Objective To investigate the relationship between urinary iodine level and thyroid disease.Methods The study used a case-control design.One hundred and nine patients with thyroid disease from the Affiliated Hospital of Shanxi Institute for Endemic Disease Control were selected as case group from 2011 to 2012,and these patients were divided into three groups:Graves's disease (GD) group (n =48),chronic lymphocytic thyroiditis (HT) group(n =34) and thyroid nodules group(n =27).Sixty-two healthy people from the same region were selected as a control group.Urinary iodine was determined using arsenic cerium catalytic spectrophotometry,thyroid autoantibody (TRAb) and thyroid peroxidase antibody (TPOAb) was detected using electrochemiluminescence,while iodine absorption rate was measured using thyroid function analyzer,and thyroid volume was measured using type-B ultrasonic method.The relationship between urinary iodine level and patients with thyroid disease was compared with that of control group.Results Urinary iodine levels of patients with GD,HT,thyroid nodules and control groups were 313.95,375.20,220.20 and 196.50 μg/L,respectively.Urinary iodine levels of patients with GD and HT groups were higher than that of control group(Z =3.238,4.275,all P ＜ 0.0125).Urinary iodine level of patients with HT was higher than that of thyroid nodules(Z =3.762,P ＜ 0.0125).Iodine uptakes of GD,HT,thyroid nodules and control groups were (84.20 ± 16.90)％,(23.51 ± 6.72)％,(28.34 ± 8.02)％ and (29.31 ± 8.41)％; TRAbs of patients with GD,HT,thyroid nodules and control groups were (58.57 ± 20.31)％,(2.54± 1.00)％,(2.98 ± 0.83)％ and (3.01 ± 1.21)％; TPOAbs of patients with GD,HT,thyroid nodules and control groups were (117.03 ± 57.21)％,(251.00 ± 98.20)％,(16.81 ± 9.87)％ and (15.00 ± 7.23)％.Iodine uptake,TRAb and TPOAb of GD group were higher than those of control group(P ＜ 0.05).TPOAb of HT group was higher than that of control group(P ＜ 0.05).Urinary iodine levels of GD group and HT group were positively correlated with TPOAb(correlation coefficient were 0.462,0.478 all P ＜ 0.05).Conclusions Excessive iodine intake is found in patients with GD and HT.Determination of urinary iodine is helpful for individualized iodine supplementation.

In this study, 17 kinds of Dendrobium species of Fengdous including 39 individuals were collected from 4 provinces. Mitochondrial gene sequences co I, nad 5, nad 1-intron 2 and chloroplast gene sequences rbcL, matK amd psbA-trnH were amplified from these materials, as well as nrDNA ITS. Furthermore, suitable sequences for identification of Dendrobium species of Fengdous were screened by K-2-P and P-distance. The results showed that during the mentioned 7 sequences, nrDNA ITS, nad 1-intron 2 and psbA-trnH which had a high degree of variability could be used to identify Dendrobium species of Fengdous. However, single fragment could not be used to distinguish D. moniliforme and D. huoshanense. Moreover, compared to other combined fragments, new type combined fragments nrDNA ITS+nad 1-intron 2 was more effective in identifying the original plants of Dendrobium species and could be used to identify D. huoshanense and D. moniliforme. Besides, according to the UPGMA tree constructed with nrDNA ITS+nad 1-intron 2, 3 inspected Dendrobium plants were identified as D. huoshanense, D. moniliforme and D. officinale, respectively. This study identified Dendrobium species of Fengdous by combined fragments nrDNA ITS+nad 1-intron 2 for the first time, which provided a more effective basis for identification of Dendrobium species. And this study will be helpful for regulating the market of Fengdous.
DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Dendrobium ; classification ; genetics ; Genes, Chloroplast ; Genes, Plant ; Introns ; Plants, Medicinal ; classification ; genetics

AIMTo determine the involvement of NO signal pathway in the development of hyperalgesia induced by activation of protein kinase C (PKC ), nociceptive responses and nitric oxide synthase(NOS) expression and nitric oxide (NO) content in the spinal cord were observed after administration of Phorbol 12-Myristate-Acetate (PMA), a PKC agonist, in rats.

METHODSNociceptive response was observed by behavioral approach. Pain threshold was assayed using thermal tail-flick test. NADPH-d histochemistry was used to investigate the changes of NOS expression. Nitrate/nitrite (NO3-/NO2-) was assayed to represent NO content of lumbar enlargement of spinal cord.

RESULTSNociceptive response was induced and pain threshold decreased after intrathecal injection of PMA. The number of NADPH-d positive cells increased significantly in the superficial layer of the spinal cord dorsal horn (Laminae I - II ) and the grey matter surrounding the central canal (Laminae X), and the reactive degree of NADPH-d positive soma and processes and NO content of the lumbar enlargement of the spinal cord increased significantly after intrathecal injection of PMA. Pretreatment of PKC inhibitor chelerythrine chloride blocked the changes induced by PMA.

CONCLUSIONThe activation of PKC in the spinal cord neurons might induce spontaneous nociceptive responses and hyperalgesia in rats, as well as promote NOS expression and NO production, suggesting that increase in NO production is one of mechanisms of hyperalgesia induced by activation of PKC.

Animals ; Enzyme Activators ; pharmacology ; Nitric Oxide ; biosynthesis ; Nociception ; drug effects ; Pain Threshold ; drug effects ; Protein Kinase C ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spinal Cord ; drug effects ; metabolism ; Tetradecanoylphorbol Acetate ; analogs & derivatives ; pharmacology

OBJECTIVETo investigate the effect of hepatocyte growth factor (HGF) on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) after allogeneic bone marrow transplantation (allo-BMT) and related mechanism in acute lymphoblastic leukemia (ALL) mice.

METHODSTwenty nude mice were randomly divided into control (group A) and test (group B) groups for monitoring relapse, and 20 BALB/c mice into control (group C) and test (group D) groups for GVHD. HGF as injected from day 0 to day 7 after BMT for groups B and D, while PBS for A and C. CD4(+) and CD8(+) T cell were evaluated by flow cytometry. The survival of mice after BMT was recorded. The level of tumor necrosis factor-alpha (TNF-alpha) was evaluated by ELISA.

RESULTSThe median past-BMT survival were 7.00 +/- 1.58, 9.00 +/- 1.58, 11.00 +/- 3.95 and 24.00 +/- 13.44 days for groups A, B, C, D, respectively, being prolonged in group D. HGF could decrease the quantity of CD4(+) T cells [group D (10.39 +/- 1.15)% vs group C (13.50 +/- 1.80)%, P < 0.01] and increase CD8(+) T cell [group D (12.25 +/- 2.85)% vs group C (6.12 +/- 1.99)%, P < 0.01], decrease the level of TNF-alpha in transplanted ALL mice [group D (112.10 +/- 18.99) pg/ml vs group C (143.90 +/- 25.35) pg/ml, P < 0.01] and reduce the degree of GVHD.

CONCLUSIONHGF could alleviate post-allo-BMT GVHD but retain GVL effect.

Animals ; Bone Marrow Transplantation ; Disease Models, Animal ; Female ; Graft vs Host Disease ; prevention & control ; Graft vs Leukemia Effect ; drug effects ; Hepatocyte Growth Factor ; pharmacology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; surgery ; Random Allocation ; Transplantation, Homologous

Purpose To observe the effect of Necrostatin-1 (Nec-1),a necroptosis inhibitor,on the inflammation in unilateral ureter obstruction mice.Methods Male C57BL/6J mice were randomly divided into sham operation group,unilateral ureter obstruction operation group and UUO + Nec-1 treatment group,and the mice were sacrificed at 7th day after operated.Scr and BUN were measured.The pathological changes in the kidney were observed by HE staining.Immunohistochemistry and Western blot were used to detect the protein expression of necroptosis-related indicators RIP1,RIP3,MLKL and inflammatory cytokines TNF-α,IL-1β and MCP-1.Results Compared with sham operation group,the expression of RIP1,RIP3 and MLKL protein increased in the renal tissue of UUO mice,accompanied with increased expression of TNF-α,IL-1β and MCP-1.Nec-1 treatment significantly decreased above-mentioned protein expression in UUO mice,and also reduced renal interstitial inflammation and renal tubal injury according to HE staining.Scr and BUN levels suggested improved renal function.Conclusion Nec-1 could relieve the inflammatory reaction in renal tissue of the UUO mice by inhibiting necroptosis,which may be a new target for the treatment of secondary inflammation.

Single-Strand Conformation Polymorphism(SSCP) is an effect method for investigating environment microbial genetic polymorphism, with its characterization of rapidness, simplicity, and sensitivity. However, many factors can influence the results of SSCP in the analysis of complex environment samples, and its optimization is highly needed. In this paper, optimal PCR-SSCP conditions were discussed based on PAGE concentration, formamide deionized in denaturing loading buffer, electrophoresis time and temperature. The resluts showed that the optimal conditions were as follows: 16S rDNA V1~V3 was selected as the targeted gene, the ratio of acrylamide to N, N-dimethylacrylamide in 12% polyacrylamide gel electrophoresis(PAGE)gel was 49:1, the ratio of formamide deionized in denaturing loading buffer was 1:3, running the SSCP gel at 300 V for 18 h (under 4 ℃). Aside from this, the validations using samples from a simultaneous desulfurification and denitrification bioreactor were conducted under this optimal conditions.

Objective:To develop a novel regulable sutureless aortic arch prosthesis and to apply it in an animal experimental study.Methods:The arch skeleton of the prosthesis was made of tandem Z-shape NiTiNOL wire;the branch skeleton was made of laser-cut NiTiNOL tube;and the whole skeleton was coated with thin ePTFE film.The blood vessel was anastomosed by di- rect ligature,needing no manual suturing.The prosthesis was applied in swine aortic arch operations under the bypass condi- tion.The practicality for surgery and the feasibility of anastomosis of the prosthesis were assessed.Results:Aortic arch opera- tions were successfully performed in 6 of the 8 experimental animals.The prostheses were easy to use,and the mean bypass time was only 10 min.The blood loss of the anastomoses was less than 100 ml within 8 h postoperatively in 5 animals;one had more blood loss due to prosthesis mismatch.Conclusion:The novel regulable sutureless aortic arch prosthesis has satisfactory practicality for surgery and reliable anastomosis,making it promising in future clinical application.