Objective To investigate the mechanism of glycoprotein serglycin (SRGN) promoting metastasis of breast cancer cells and the possible mechanism of SRGN expression.Methods Real time quantitative polymerase chain reaction (PCR) and bioinformation retrieval were used to detect the expression of SRGN in lymph node metastasis and non-metastasis breast cancer.MDA-MB-231 shRNA and MCF-7-SRGN of breast cancer stable cell line were established by lentivirus shRNA interferencc and overexpression.Transwell assay was used to test the effect of SRGN on invasion and metastasis of breast cancer cell line in vitro.Western blot assay was used to detect the changes of epithelial-mesenchymal (EMT) related markers.The possible regulatory mechanism of SRGN expression was detected by Western blot assay.Results SRGN expression was significantly increased in lymph node metastasis of breast cancer in clinical specimens.SRGN interference inhibited the invasion and metastasis of tumor cells.SRGN promoted breast cancer cells EMT.Transforming growth factor β1 (TGFβ1) promoted the expression of beta SRGN transcription.Conclusions SRGN can induce the change of EMT in breast cancer cells and promote the invasion and metastasis of breast cancer cells.

Objective:To investigate the clinical significance of LncRNA anti-differetiation non-coding RNA (ANCR) expression in tumor tissues of gastric cancer patients and its biological effects on cells.Methods:72 cases of gastric cancer tissues and corresponding adjacent tissues were collected from Sep. 2016 to Jun. 2018 in our Hospital. Gastric cancer cell HGC-27 was cultured, lentiviral transfected ANCR cDNA full-length vector was used as a Test group in HGC-27 cells, and transfected blank vector as a control group. Real-time quantitative PCR (qPCR) was used to detect the expression of ANCR, transcription factor Oct4 and Sox2 mRNA in tissues or cells, Western blot was used to detect the expression levels of Oct4 and Sox2 in cells, CCK-8 assay was employed for detecting cell proliferation in both groups, and Transwell invasion and migration assay was used to detect the transfer ability of cells in the two groups.Results:The expressions of ANCR in gastric cancer and corresponding adjacent tissues were respectively 0.013 (0.006, 0.025) and 0.041 (0.011, 0.136) , and the expression of ANCR in gastric cancer tissues was significantly higher than that in adjacent tissues ( P<0.01) , and patients with high expression of ANCR had higher TNM stage and lower cell differentiation ( χ2=7.414 and 8.236, P<0.05) . The expressions of ANCR mRNA in control group and test group were respectively 1.000±0.064 and 6.250±0.889, Oct4 mRNA were respectively 1.000±0.208 and 2.815±0.349, Sox2 mRNA were respectively 1.000±0.173 and 2.526±0.390, Oct4 protein were respectively 1.000±0.148 and 3.396±0.105, Sox2 protein were respectively 1.000±0.119 and 2.916±0.130, and the expressions of ANCR, Oct4 and Sox2 mRNA in the test group were significantly higher than those in the control group ( P<0.01) ; the expression levels of Oct4 and Sox2 protein in the test group were significantly higher than those in the control group. The proliferation abilities of control group and test group were 7.164±0.426 and 9.627±0.605 in 72h, and 13.750±1.089 and 19.166±1.649 in 96h. The proliferation of cells in the Test group at 72 and 96 hours was significantly higher than that in the control group ( P<0.01) . The average number of invasive cells per visual field in control group and test group were 17.26±5.48 and 39.43±5.21, and number of migration cells were 30.49±7.74 and 62.20±7.51, and the number of migration and invasion cells in the Test group was significantly larger than that in the control group ( P<0.01) . Conclusions:The expression of LncRNA ANCR in tumor tissues of gastric cancer patients is significantly increased, and it is closely related to the progression of the disease of patients and the degree of cell malignancy. It can promote the expression of gastric cancer stem cell markers in vitro and enhance the ability of cell proliferation and metastasis.

AIM To establish anti-osteosarcoma antibody producing hybridoma cell lines and to study the characterization of the monoclonal antibodies. Methods BALB/c mice were immunized with human osteosarcoma cells OS-9607 and the immunized spleen cells were fused with SP2/0 cells to raise hybridoma. The propert of antibody and it's cytotoxic effect were studied respectively with immunohistochemistry methods using OS-9607 and normal hepatocytes、 Western Blot methods and MTT method. Results A hybridoma cell line named 3D9 was established and it secreted high quality mAbs steadily. 3D9 cell had all the characteristics of hybridoma. The mAb's corresponding antigens was specifically and highly expressed in human osteosarcoma. With enzyme-labeled immunohistochemical staining on formaldehyde -fixed sections from human osteosarcoma,it was found that 83％ of the specimens expressed the corresponding antigen. Most of them were expressed on the nuclear of cells, no positive expression was observed in kinds of normal tissues. Western Blot showed 3D9's corresponding molecule weight is Mr54 000. MTT assay proved that the cytotoxicitis of effective groups were higher than control groups. Conclusion A high quality hybridoma is cultured and the mAb secreted by it has osteosarcoma specificity and obvious cytotoxic effect. It may be a new biochemical mark of osteosarcoma, and it's clinical prospect of immunotherapy will be wide.

Objective To explore the age-and sex-specific body composition of normal children in Beijing area.Metheds The subjects were a total of 587 children of 6-14 years old,who were recruited from Beijing schools.All of them had relative weight within normal range(80%～120%),and no chronic disease.The relative weight was obtained,according to standard weight,using the follo-(wing) formula: relative weight(%)=(body weight/standard weight) ?100.Body compositions were estimated with a bioelectrical impedance analyser,which had been proved to be reliable and valid for determining the percentage of body fat.Results Not only fat free mass(FFM) but also fat mass(FM) increased monotonically with age in both sexes.FFM was higher in boys than that in girls at all ages.FM was significantly higher in girls than that in boys aged 6 to 8 years old;however,there was no significant difference for FM between sexes aged 9-14 years old.Patterns of change in mean ratio of body fat(%BF),with age differed by sex.Percent age of BF was significantly higher in girls than that in boys at all ages except at 10 and 11 years old. In boys,%BF increased with age,while in girls it remained nearly constant from age 6 to 10 years old,and gradually increased from age 10 to 14 years old.Body mass index(BMI) increased steadily with age in both sexes,and boys had consistently higher BMI than girls.In boys,the increase in BMI was steeper from age 10 to 14 years old.Even in the subjects with BMI

0.05).Conclusion pcDNA3-gB with different doses have not significant effect on the indexes of hematology,hematological biochemistry and pathology in immunized mice.It is initially proved that pcDNA3-gB is safe.

Objective To compare the clinical effects and safety of surgical clipping and intravascular interventional therapy in treatment of intracranial wide-necked aneurysm.Methods The clinical data of 158 patients with intracranial wide-necked aneurysm from February 2010 to February 2013 were retrospectively analyzed,all patients were divided into two groups:surgical clipping group with 92 cases and intravascular interventional therapy group with 66 cases,the postoperative curative effects,treatment time,hospital stay,hospital expenses and postoperative complications between two groups were compared.Followed up for 10-46 months,the recurrence rate were compared.Results The good prognosis and defective rates between surgical clipping group and intravasular interventional therapy group had no significant difference [90.2％(83/92) vs.90.9％(60/66),9.8％(9/92) vs.9.1％ (6/66)] (x2 =0.298,P ＞ 0.05).The preoperative Hunt-Hess classification and CT Fisher classification between two groups had no significant difference (P ＞ 0.05).Six months after discharge,mRS score was used to evaluate the curative effect,the defective rates in same level patients between two kinds of treatment methods had no significantdifference (P ＞ 0.05).The treatment time,hospital stay in surgical clipping group were significantly longer than those in intravascular interventional therapy group [(4.03 ± 1.01) h vs.(1.61 ± 0.98) h,(15.90 ± 2.03) dvs.(13.20 ± 1.95) d],hospital expenses was significantly lower than that in intravascular intervention therapy group [61 829.4 ±320.6) yuan vs.(99 876.2 ±371.5) yuan] (P ＜0.05).The postoperative complications rate between two groups had no significant difference (P ＞ 0.05).Followed up for 31.3 (10-46) months,the recurrence rate in surgical clipping group was significantly lower than that in intravascular intervention therapy group [1.1％ (1/94) vs.8.8％ (6/68)] (P ＜ 0.05).Conclusion Surgical clipping and intravascular interventional therapy in treatment of intracranial wide-necked aneurysm has their own different characteristics,so patients' treatment methods should be based on their preoperative status (especially preoperative Hunt-Hess and Fisher classification) and patients' economic conditions.

Endothelialprogenitorcels(EPCs)arethepluripotentstemcelsofvascularendothelial cels. They have self-differentiation and proliferation ability. A large number of animal experiments and preliminary clinical studies have show n that EPCs have broad prospects of clinical application. This article review s the research status of EPCs and their application in the clinical treatment of ischemic stroke.

Objective To compare the occurrence of postoperative cognitive dysfunction (POCD) in elderly patients using differentanesthetic methods.Methods Ninety-three ASA Ⅱ or Ⅲ patients,aged ≥ 65 yr,weighing 45-67 kg,scheduled for artificial femoral head replacement,were randomly divided into 2 groups:general anesthesia (group G,n =47) and combined spinal-epidural anesthesia group (group S-E,n =46).In group G,anesthesia was induced with iv injection of midazolam 0.1 mg/kg,propofol 2 mg/kg,fentanyl 3-5μg/kg,and vecuronium 0.1 mg/kg,and maintained with continuous infusion of propofol 2-3 mg· kg-1 · h-1,intermittent iv boluses of fentanyl 1 μg/kg and vecuronium 0.04 mg/kg and inhalation of 1.5％-2.0％ isoflurane.In group S-E,hyperbaric 0.5 ％ ropivacaine 2 ml was injected into the subarachnoid space over 20 s,the patients were kept in the original position for 15 min,the level of anesthesia was simultaneously adjusted to below T8 on the operated side,and 0.5 ％ ropivacaine 3-5 ml was injected into the epidural space when needed during operation.Cognitive function was assessed by mini-mental state examination at 24 h before anesthesia and 24 and 72 h after operation.Venous blood samples were collected for determination of plasma amyloid-beta levels by ELISA.Results Compared with group G,the incidence of POCD at 24 h after operation and level of plasma amyloid-beta were significantly decreased in group S-E (P ＜ 0.05).Conclusion Elderly patients are more likely to develop POCD under general anesthesia than under combined spinal-epidural anesthesia.

AIM To study the pharmacokinetics of arbidol capsule in Chinese healthy volunteers.METHODS A single oral dose of arbidol capsule 200 mg was given to 20 healthy volunteers respectively.Plasma samples were prepared based on a simple liquid-liquid extraction.The extracted samples were analyzed by HPLC equipped with UV detection.Pharmacokinetic parameters were calculated by 3P87 software. RESULTS The main pharmacokinetic parameters of arbidol were as follows:c(max)(418±s 241)μg·L-1,t(max)(1.3±1.2)h,t(1/2α)(1.9±2.3)h,t1/2β(14±5),hAU0-t(2 633±1 071)μg·L-1,Vc/F(0.7±0.6)L,CL(0.08±0.03)L·h-1,CONLUSION The pharmacokinetics of arbidol capsule in human body accord with two-compartmetn open model.The study will offer the pharmacokinetic parameters for the clinical application of arbidol.

HIV-1 integrase (IN) is a key enzyme for the viral replication. The protein-protein interaction (PPI) between HIV-1 IN and a cellular cofactor lens epithelium-derived growth factor (LEDGF/p75) is a validated target for anti-HIV drug discovery. In order to build the platform for screening inhibitor against PPI between IN and LEDGF/p75, the vector containing the LEDGF/p75 protein cDNA was constructed and expressed in Escherichia coli and the function of the LEDGF/p75 protein was assayed. The LGDGF/p75 encoding gene optimized according to the preference codon usage of E. coli, was synthesized and cloned into the expression vector pGEX-4T-1 to form a recombined plasmid, then transformed into host cell E. coli BL21 (DE3). The recombined clones were identified and confirmed by BamH I/Sal I digestion and sequencing, the successfully recombined plasmid in the host cell was induced by IPTG and the condition of the expression was optimized. The expressed protein was purified by the Ni2+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombinant protein. The recombinant LGDGF/p75 was soluble, and expressed highly and stably in E. coli. The protein was proved to enhance HIV-1 IN strand transfer activity in vitro by ELISA. It will be helpful to build the platform of screening inhibitors against PPI between IN and LEDGF/p75.
Cloning, Molecular ; Escherichia coli ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; physiology ; Humans ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Protein Binding ; Virus Replication