HIV Infections ; prevention & control ; Humans

Objective To establish and verify a multiplex fluorescence quantitative PCR method for detection of CHO host cell residual DNA content and fragmentation degree, so as to provide a reliable method for safety detection of related vaccines.Methods Two sets of primers(Alu-L and Alu) and universal probe were designed for clone250 and clone49c, members of Alu sequence family of CHO cells. The UBI3 gene of capsicum was introduced as external standard gene, and corresponding primers and probe were designed. Multiplex reaction system of Alu sequence and external standard gene was established, and the residual DNA of CHO cells in samples was quantified by the multiplex fluorescence quantitative PCR. The linear range,limit of quantitation(LOQ), specificity, robustness, precision and accuracy of the method were verified. In addition, the DNA fragmentation degree of CHO cells was determined by comparing the △Ct [Ct(Alu-L)-Ct(Alu)] of the amplification curves of long and short primers of Alu sequences. Results The multiplex reaction standard curve had a good linear relationship in the range of 0. 001-0. 1 ng/μL, R~2≥ 0. 99. The addition of external standard gene had no effect on the detection ability for CHO target, and the LOQ was 0. 5 fg/μL. The DNA of human, E.coli, rats and mice had no effect on the detection, the primers targeting residual DNA of CHO cells had no specific amplification in the genomes of seven animal cell lines, and the fragmented DNA samples showed no effect on the detection results. The CVs of precision verification were all less than 15%, and the recoveries of simulated samples in normal saline and PBS containing 1% BSA were in the range of 70%-130%. The△Ct of Alu and Alu-L amplification curves increased with the degree of sample fragmentation, and the fitting curve R~2≥ 0. 99.Conclusion The multiplex fluorescence quantitative PCR detection method established in this study can rapidly and accurately quantify the residual DNA of CHO cells and determine the fragmentation degree of samples according to △Ct, which can be used for the safety detection of the related vaccines.

Liquichip technology utilizes different fluorescent coding microbead as carriers of bio-probes and flow cytometry as optical detection method,to accomplish bio-molecules reaction in suspension liquid system.This article introduces the application of liquidchip technology in the analysis of autoimmune disease,cytokine,contagious disease,endocrinopathy,neural disease,tumor marker detection and so on.With the advantages of high sensitivity,high speed,high flux,multi-analyte,high accuracy and excellent repeatability,liquichip technology has promising prospect to be widely used as advanced biomedical detection platform.

Expansive type of hepatocellular carcinoma (HCC) is a subtype of HCC. Correlation between the preoperative sonography and postoperative pathology was studied in 35 cases of HCC. According to the classification of nakashima, 21 cases belonged to the expansive type whereas the other 14 cases the infiltrative type. Different from the infiltrative type. the expansive type of HCC had four specific sonographic appearances: hypoechoic ring, hypoechcic halo, pedunculated and small nodular echo which had different pathological bases. Expansive tyje of HCC was not so commonly seen as infiltative type but its rate of surgical resection was mu~h higher than that of infiltratine type (JP< 0. 01). Thus, it is the kind of HCC with significant clinical importance. Preoperative ultrasonographic diagnosis help to decide the way of treatment.

HLA (human leukocyte antigen) is critical molecule relevant to immunoresponse, which is always involved in antigen presentation and determining killing activity of immune cells against tumor. The abnormal expression of HLA in lung cancer is believed to be a common phenomena of tumor biology. It has been demonstrated that HLA expression varies in various types of lung cancer, even in differential stages of one type of lung cancer. HLA expression is highly associated with genre, progression and prognosis of lung cancer. The thorough investigations of the relationship between HLA and lung cancer and of mechanism of HLA abnormality will provide useful information for studying of development of lung cancer and tumor immunotherapy.

This study aimed at investigating the effects and mechanisms of 7-O-succinyl macrolactin A in inhibi-ting human lung cancer. After treatment of human lung cancer cell lines H460 with 7-O-succinyl macrolactin A, MTS assay was employed to determine cell proliferation;crystal violet staining was used to detect cell adhesion of H460;transwell chamber assay and wound healing assay were performed to evaluate cell invasion and migration;and flow cytometry assay was adopted to evaluate cell cycle. Western blotting and real-time PCR were also employed to determine the expression of β-catenin, c-Myc, Cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2 , Survivin and MMP-2/9. The phosphorylation of AKT and mTOR was determined as well. In vitro prolifera-tion of H460 was inhibited significantly by 7-O-succinyl macrolactin A. Cell adhesion, invasion and migration abilities were also attenuated. Western blot and real-time PCR showed that the expressions of β-catenin, c-Myc, cyclin D1, vimentin, N-cadherin, CD44, integrin β1, Bcl-2, Survivin and MMP-2/9 were down-regulated by 7-O-succinyl macrolactin A. It was also found that phosphorylation of AKT and mTOR was inhibited by 7-O-succinyl macrolactin A. 7-O-succinyl macrolactin A can inhibit the in vitro growth and invasion of human lung cancer cell lines H460.

Spine metastasis stereotactic body radiation therapy delivers conformal high radiation dose to the tumor target, and steep falloff dose gradients protect adjacent normal structures, especially spinal cord. Some studies demonstrated stereotactic body radiation therapy for metastatic tumors of the spine is safe and clinically effective. This technique markedly improved local control with an excellent symptomatic response compared with conventional external beam radiation therapy techniques, and is beginning to change the treatment paradigms for metastatic spine disease both as post-operative adjuvant therapy and primary definitive local treatment. Although the technique is being increasingly used in clinical practice, the optimal fractionation schedule remains uncertain. Based on the research of radiobiologic principles, radiological physics and clinical data, multiple session stereotactic body radiation therapy has some advantage in clinic.

Objective To investigate the methods of preoperation diagnosis and treatment of primary gastric lymphoma of mucosa-associated lymphoid tissue(MALT).Methods To analyze retrospectively the diagnosis and treatment of 54 cases of MALT lymphoma diagnosed surgically and pathologically.Results The most common symptoms were stomachache or discomfort.Of the patients with MALT lymphoma,85.2%(46/54)cases were diagnosed by preoperation gastroscopy with biopsy.Helicobacter pylori was found in 48 cases(88.9%).The overall resection rate for MALT lymphoma was 87%(47/54).The 5-year and 10-year survival were 77.1% and 72.9%,respectively.Conclusions It is obvious that MALT lymphoma is associated with Helicobacter pylori.Since no specific symptoms in gastric MALT lymphoma,the preoperative diagnosis is relied upon the application of gastrointestinal image,gastroscopy and repeated biopsy.The use of curative resection with the post-operative chemotherapy and/or radiotherapy to treat MALT lymphoma is reasonable.

Aim To study whether or not the green tea catechins(EGCG)has physiological benefits and the underling protective mechanism.Methods The subunit protein levels of ?4、?7 of nAChR were detected by BCA protein assay,Dot Blot assay and MTT assay.Results The results showed that the green tea catechins can significantly reduce the subunit protein levels of nAChR and decrease the cell activity induced by A?1-40.Conclusions EGCG can provide neuroprotection in vitro by up-regulating nAChR sununit levels and inhibiting the neurotoxin of A?1-40.

Objective To study the inhibitory effect of yan xiao capsules on hepatocarcinoma mice. Methods Forty mice were randomly divided into control group ,CTX group,ADM group,trial group. Observe tumour weight ,the span time of tumour-bearing mice, tumour morphology and the mortality rate of ascites cancer cell by direct killing respectively. Results The tumour weight markedly decreased in trial group, the rate of inhibition tumour was 52.0 %(P