BACKGROUND:Matrix metaloproteinase-1 can degrade extracelular matrix, which is mainly colagen type I, and has the potential to reverse fibrosis tissue. OBJECTIVE:To construct the recombinant adenovirus vector containing human matrix metaloproteinase-1 (hMMP-1) gene with GatewayTM Clone Technology, and observe the capacity of degrading colagen type IIIin vitro. METHODS: The gene hMMP-1 was amplified by using PCR from the pcDNA3.1 plasmid and was cut down by the double endonuclease. The linear gene fragment was connected to the entry vector pENTERTM 1A. Then the entry clone and the destination vectors pJTI? R4 Dest CMV-N-EmGFP pA Vector recombined using the LR reaction to form the expression clone pAd-hMMP-1-eGFP. The linear pAd-hMMP-1-eGFP cut down by endonucleasePac I was transfected into HEK293A cels to packaging the Ad-hMMP-1-eGFP. The transfected situation was observed under a fluorescence microscope, the target protein expression was detected by western-blot assay and RT-PCR. Cels can be divided into three groups: blank control group: HEK293A cels, AD-EGFP group: HEK293A cels were infected by Ad-eGFP, AD-HMMP1-EGF group: HEK293A cels were infected by Ad-hMMP1-eGFP and colagen type III. The content of colagen type III was detected by ELISA kits after 24, 48 and 72 hours. RESULTS AND CONCLUSION: It was confirmed that the entry vector and the destination vector both contained hMMP-1 target gene by restriction analysis and sequencing. The green fluorescent protein was observed in the 293A cels transfected by the Ad-hMMP-1-eGFP at 4 days. The fluorescence intensity was the highest at 10 days. The virus was colected at 12 days, the viral titer was determined as 4.84 × 1010 PFU/mL, the target protein was efficient expressionvia western-blot assay. Blank control group and AD-EGFP group had no obvious change of colagen content with the extension of time. The rate of colagen degradation in AD-HMMP1-EGFP group was 24%, 56% and 81% respectively at 24, 48, 72 hours. AD-HMMP1-EGFP group degraded colagen significantly compared with the other two groups (P < 0.01). The recombinant adenovirus vector containing hMMP-1 was successfuly constructed by using the Gateway technology, this method was more efficient and specific than with the traditional methods. The hMMP1 degraded colagen type III significantlyin vitro.

Objective To detect and explore the expression of ARD1 and its clinical significance in the nasopharyngeal in-flammatory tissue ,nasopharyngeal carcinoma group and its subgroups .Methods Expression of ARD1 in nasopharyngeal carcinoma (56 cases) and nasopharyngeal inflammatory tissue (20 cases) were detected by immunohistochemical staining SP ,the correlation between the expression of ARD1 and age ,gender ,histological grade ,TNM clinical stage and tumor metastasis were analysed . Results The positive expression rate of ARD1 were 10 .00% (2/20) ,55 .35% (31/56) in the nasopharyngeal inflammatory tissue and nasopharyngeal carcinoma ,respectively .The expression level of ARD1 in nasopharyngeal carcinoma was significantly higher than in the nasopharyngeal inflammatory tissue ,the difference was significant (P< 0 .05) ;expression of ARD1 in the nasopharynge-al carcinoma was correlated with the histological grade of nasopharyngeal carcinoma(P< 0 .05) and the expression was increased in poor differenciation tissue .But there was no statistical difference between the expression of ARD1 and the patient′s age ,gender , TNM clinical stage ,tumor metastasis(P> 0 .05) .Conclusion The expression of ARD1 is high in nasopharyngeal carcinoma ,and has a closely correlation with diffferentiation level of tumor ,which suggested that ARD1 may be involved in the the occurrence and development of nasopharyngeal carcinoma .However ,further research needs to be done for its mechanism in the nasopharyngeal car-cinoma .

Two patients underwent liver transplantation of blood type incompatibility were collected from Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA. Case 1: A male who had primary hepatic carcinoma underwent classic orthotopic liver transplantation; the blood of donor was type A, and the blood of recipient was type O. Case 2: A female having history of type B hepatitis underwent classic orthotopic liver transplantation due to pregnancy combining with severe liver disease and coagulation disorder; the blood of donor was type B, and the blood of recipient was type O. Immunohistochemistry staining was used to observe pathological changes and deposition of various immunoglobulin and complement in two cases following liver transplantation of blood type incompatibility under optic microscope and to investigate diagnostic standard of humoral rejection. The results showed that linear or granular depositions of IgG, IgM, IgA, C4c, C4d, and Clq were found in endothelial cells of hepatic sinusoid, suggesting that IgG and other immunoglobulin exhibited a strongly positively diffused deposition on the endothelial cells of hepatic sinusoid, while expression of C4d and other complements was also found. All those mentioned above could be considered as evidences to prove onset of humoral rejection in transplanted liver tissue.

BACKGROUND: CpG oligodeoxynucleotide (ODN) is a type of highly effective immune adjuvant with low toxicity, which has an extensive application in gene therapy for many diseases. However, the specificity for species and cells leading to low uptake by cells and degradation by nuclease blocks its clinical application. OBJECTIVE: To explore the specific delivery and its immunologic efficacy of CpG ODN targeting B lymphocytes of umbilical cord blood by CD40 ligand-receptor-mediated carrier system. DESIGN, TIME AND SETTING: An observation and control experiment was performed at the Department of Hematology, and Department of Pediatric, the First Affiliated Hospital of Jinan University from April 2004 to October 2007. MATERIALS: Fresh umbilical cord blood with heparin was obtained from healthy, natal infant. Informed consent was obtained from his parents, and the experiment was approved by the hospital Ethics Committee. METHODS: CD40 ligand (CD40L)-EDC-PLL-CpG ODN conjugated complex was prepared. Mononuclear cells (MNCs) from umbilical cord blood were co-cultured with conjugated complexes. Uptake rate, mean fluorescence intensity of FAM marked CpG ODN, expressions of MNCs, proliferations of lymphocytes and the IgG levels of culture supematants were detected by flow cytometry, fluorescence techniques, MTT assay and ELISA, respectively. MAIN OUTCOME MEASURES: The uptake rate, the mean fluorescence intensity of CpG ODN by MNCs, subgroups and proliferations of lymphocytes, and IgG levels of culture supematants. RESULTS: Compared to the pure CpG ODN group, the uptake rate of the conjugated complexes group was higher (98%), the peak level of up-taking occurred earlier, and intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, and CD20+ was increased, A value and IgG levels in supematants were all higher than that of the control group. CONCLUSION: CD40 tigand-receptor-mediated carrier system is helpful for CpG ODN delivery targeting to B lymphocyte, enhancing its immunological efficiency.

98%), the peak level of uptake occurred earlier, intracellular fluorescence intensity maintained much more stable. Expressions of CD19+, CD22+, CD20+ increased significantly. A_~570 values of MNCs proliferation and IgG levels in supernatant were all higher. CONCLUSION: CD40 ligand-PLL carrier system may delivery CpG ODN targeting to B lymphocytes, enhancing its immunological efficiency.

Objective To analyze the advantages and disadvantages of minimally invasive video -assisted thyroidectomy(MIVAT)and totally endoscopic thyroidectomy (TET).Methods The study included 70 patients undergoing MIVAT and TET at Qilu Hospital of Shandong University and Yantai Yuhuangding Hospital from Jan . 2012 to Dec.2012.46 patients undergoing MIVAT(MIVAT group)and 24 patients undergoing TET(TET group) were analyzed retrospectively .Data of gender,age,operative data,intraoperative and postoperative complications , length of stay , feelings of patients and cosmetic effect were collected and analyzed .MIVAT and TET were com-pared in terms of safety ,trauma and cosmetic effect .Results ①About safety:no permanent recurrent laryngeal nerve(RLN)palsy and parathyroid injury occurred in either group .One case in MIVAT group and no case in TET group was converted to open surgery (2.2%vs 0%,P>0.05) .②About trauma:the intraoperative blood loss and postoperative drainage amount in the MIVAT group were significantly less than those in the TET group ( P <0.05).Postoperative VAS score at 6 hours was similar,but MIVAT group showed lower postoperative VAS score than TET group at 12 hours and 24 hours ( P<0.05 ) .No significant difference was observed between the 2 groups about C reactive protein (CRP)on the 1st day before operation,the 1st and 3rd day after operation(P>0.05).③About cosmetic effect:the 2 groups had similar patient satisfaction index .Conclusions The study shows similar results in safety and trauma .The 2 approaches are both safe and valid .Patients using the two ap-proaches are equally satisfied with the cosmetic effect .

AIM: To detect the expression of cytoplasmic inhibitor of apoptosis protein 2 (c-IAP2) and growth arrest-specific gene 1 (GAS1) in Hodgkin's lymphoma (HL) and anaplastic large cell lymphoma (ALCL) and to investigate the role of two genes in the pathogenesis of HL and ALCL.METHODS: HE staining,the antibodies CD30,CD15,CD45RO and CD20 were used to screen the cases of HL and ALCL from 288 cases of lymphoma. The clarified HL and ALCL were subjected for immunohistochemical staining by SP and ABC methods to analyze the expression of c-IAP2 and GAS1. RESULTS: ①The positive rate of c-IAP2 in HL was 25/26(96.1%) while that in ALCL was 6/19(31.6%),there presented statistic significance between HL and ALCL groups( P

AIM: This study is based on the result of the study in HL and ALCL employing gene chip technique, in which writer found that there was distinctly different expression of caspase-4 between HL and ALCL cell lines at the level of mRNA. From the point of view, we try to identify at the level of protein whether there is different expression of this gene in HL and ALCL tissues as well. METHODS: HE staining, the monoclonal antibodies CD30 (BerH2), CD15 (C3D-1), CD20 (L26) and CD45RO (UCHL1) were used for selecting the cases of HL and ALCL. Specific high affinitive anti-caspase-4 polyclonal antibody was used by immunohistochemical staining to analyze the expression of caspase-4 in 18 cases of HL and 15 cases of ALCL. RESULTS: The expression of caspase-4 demonstrated a strong positive staining in all ALCL cases (15/15,100%), whereas negative in 16 HL cases (88 8%), while other two cases were weakly stained (11 2%), showing a distinct difference (P

Objective To investigate the clinical features of Remitting Seronegative Symmetrical Synovitis with Pitting Oedema(RS3PE),especially it’s association with malignancy. Methods The clinical data of eight RS3PE patients hospitalized during Jan 2003 and June 2007 in Peking Union Medical College Hospital were reviewed. The clinical characteristics, laboratory tests, response to treatment as well as the outcome were studied. Among the 8 patients, five were male and 3 were female,the mean age of RS3 PE onset was 60.9 (38～79yr). 5 had idiopathic RS3PE without underlying disease, and 3 presented associated neoplasm. Results All patients had symmetrical polyarthritis and pitting edema on the dorsum of hands and/or feet. Extra-articular symptoms were more conspicuous in patients complicated by neoplasm. Serum RF、AKA、APF and anti-CCP antibody were all negative. antinuclear antibodies (ANA) was positive with low titer in five cases. Hand and foot radiographs did not show bone erosion in all cases. After steroid and/or immunosuppressive treatment, 2 cases exhibited persistent remission of articular symptoms and edema, whereas relapses occurred 6, 7 and 36 months respectively after the initial diagnosis in 3 cases who had been pathologically confirmed as having Non-Hodgkin-Lymphoma and thereby received periodical chemotherapy. Conclusion RS3PE is a group of clinical manifestations with prominent heterogeneity, therefore intensive follow-up is necessary, and the presence of systemic symptoms/ signs in RS3PE patients alert the physician for the potential concomitant malignancy.

The purpose of this paper is to clarify the structure-activity relationship of anti-tumor activity of diosgenin derivatives in vitro. Study has found that diosgenin can inhibit the reproduction of tumor cells by inducing apoptosis and the main target spot of this effect is Bcl-2. Based on the characteristics of pharmacophoric points' of the three-dimensional pharmacophore for Bcl-2 inhibitors, we have docked lots of diosgenin derivatives with Bcl-2, then synthesized 31 compounds of them, finally assessed the anti-tumor activity of the diosgenin derivatives in vitro against A375, A549, HepG-2 and K562. Preliminary studies of SAR have indicated that the aliphatic esters, and aromatic esters of diosgenin without F ring have no anti-tumor activity in vitro. The triazole bromides of diosgenin all achieve fairly good anti-tumor activity in vitro, and those with larger hydrophobic group have the better activity. The stronger is the hydrogen bonding interaction and dipole-dipole interaction of the heterocyclic of diosgenin and diosgenin without F ring and the acid ester of diosgenin without F ring, the better is the activity of derivatives.