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.

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

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

Homology of three WSSV isolates, which were sampled from r epresentative maritime space of China: Tanghai isolate (Bo Bay of China), Ningbo isolate (East China Sea), Shenzhen isolate (South China Sea) was compared. Both of the genome RFLP patterns and the characteristic structural proteins SDS-PAG E electrophero grams showed that they were quite same. It suggested that they were the same ki nd of WSSV virus that caused explosive epidemic diseases of shrimps (EEDS) throu ghout southern and northern China. The same large PCR products achieved when usi ng the PCR primers from RV－PJ＝PRDV (P. japonicus, Japan) and WSBV＝PmNOBII I(P.monodon Taiwan, China) respectively to amplify the genome from P.chine nsis (Tanghai, China) with high fidelity Taq Polymerase. The sequence identiti es of WSSV from P. chinensis with those from RV－PJ＝PRDV (P.japonicus, Japan) and WSBV＝PmNOBIII (P.monodon Taiwan, China) are 97% and 100% respect ively, the results provided additional evidence that WSSV reported in different parts of the Asian and Pacific regions maybe quite the same or just different va riants of the same virus.

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.

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.

Objective To evaluate the protective effect of Yersinia pestis capsular antigen F1 and recombinant rV270 on mice after immunization with them.Methods According to body weight,40 female Balb/c mice aged 6 to 8 weeks were randomly divided into four experimental groups(Fl-10 μg + aluminum adjuvant,F1-20 μg + aluminum adjuvant,rV-10 μg + aluminum adjuvant,and rV-20 μg + aluminum adjuvant) and a control group,8 in each group.Mice in experimental groups were immunized with the natural antigen F1 and recombinant antigen rV270 adsorbed to 25％ aluminum adjuvant and the control group was immunized with the same amount of aluminum adjuvant.Each mouse was immunized at the hind leg muscle with 100 ml immunizing agent,then a booster immunization was done once on the 21st day after the first immunization.The blood of all mice was collected on the 8th week after the first immunization,serum antibody titers were detected by ELISA and the data of antibody titers were analyzed by t test for comparison between groups.At the same time the mice were injected subcutaneously with 2000-fold LD50 of Yersinia pestis virulent strain 141,after 14 days,the protective effect of immunization was analyzed.Results The control group did not produce antibody.Antibody geometric mean titers (GMT) of the F1-10 mg + aluminum adjuvant and F1-20 mg + aluminum adjuvant groups were 1 ∶ 30443.9,and 1 ∶21527.8,respectively,and compared between the two groups,the difference was not statistically significant (t =1.1282,P ＞ 0.05).The GMTs of the rV-10 μg + aluminum adjuvant and rV-20 μg + aluminum adjuvant groups were 1 ∶ 13957.3 and 1 ∶18100.9,respectively,and compared between the two groups,the difference was not statistically significant(t =0.9408,P ＞ 0.05 ).After subcutaneous injection with Yersinia pestis virulent strain 141,all mice died in the control group but all survived in the experimental group.Conclusion The immune activity of natural antigen F1 and recombinant rV270 is high,which can be used as the main component of subunit vaccine in the plague subunit vaccine study.