Objective:To construct the mammalian CO-expression plasmid pBudCE4.1-LMP-1-LMP-3,and to detect the expression of the plasmid in vitro.Methods:Fragments of LMP-1 gene and Fragments of LMP-3 gene were gained from the Company,and were constructed respectively into the plasmid vector Puc57,The inserted target genes in plasmid were verified by nucleotide sequencing and enzymes.fragments of LMP-1 gene were constructed firstly into the plasmid vector pBudCE4.1,fragments of LMP-3 gene were constructed into the plasmid vector pBudCE4.1-LMP-1 after iden tification with nucleotide sequencing and enzymes.MSCs cell line was transfected with this Co-expression plasmid using lipofectin reagent.according to the transfect situation,the MSCs were divided into 5 groups,the non-transfected group(Group A),the group transfected by empty vector(Group B),the group transfected by LMP-1(Group C),the group transfected by LMP-3(Group D)and the group transfected both LMP-1 and LMP-3(Group E).the expression of LMP-1 and LMP-3 were detected by RT-PCR and Western blot technique.Results:The plasmids Puc57-LMP-1、Puc57-LMP-3 and pBudCE4.1-LMP-1-LMP-3 were obtained successfully and verified by nucleotide sequencing and enzymes.After transfection with this mammalian Co-expression plasmid,the LMP-1 and LMP-3 molecules were expressed in MSCs cells.The results of RT-PCR and Western Blot were measured with the grey value.To the expression of mRNA and protein of LMP-1,the diferences between groups A、B and groups C、D、E were significant(P0.05);To the expression of mRNA and protein of LMP-3,the diferences between groups A、B and groups C、D、E were significant(P0.05).Conclusion:the constructed mammalian Co-expression plasmid pBudCE4.1-LMP-1-LMP-3 can express LMP-1 and LMP-3 molecules in vitro at the same time.

Objective To investigate effects of octreotide combined with aspirin on the growth and metastasis of gastric cancer in vivo. Methods SGC-7901, human gastric cancer cell line, was implanted orthotopically in the stomach of nude mice. Octreotide or aspirin alone and the combination of the two drugs were administrated for 8 weeks to investigate the effects of them on tumor growth and metastasis. Results At necropsy, xenografts in situ were found in all stomachs of nude mice except two nude mice in the combined group. The mean volume and weight of tumors in nude mice treated with octreotide or aspirin were significantly lower than those of control group. The inhibitory rate for tumors was 60.6% in octreotide group, 39.3% in aspirin group and 85.6% in the combined group with octreotide and aspirin respectively. The metastatic rates of gastric adenocarcinoma were markedly lower in any of treatment group than that in control group. No severe side action was observed in all of treatment group. Conclusion Octreotide combined with aspirin greatly enhanced the anti-growth effect on gastric cancer in vivo when compared to using either of them and inhibited the metastasis of gastric adenocarcinoma in nude mice.

Objective To investigate the effect of cytokine-induced kill cells( CIK)of the cord blood on bladder transitional cell carcinoma T24 in vitro. Methods The cord blood mononuclear cells( CBMC)were induced with different cytokines such as Interleukin 2( IL-2 ),interferon γ( IFN-γ),CD3Mc-Ab into CIK cells. The proliferation ability of CIK cells was measured by calculation in different culturing time,and the phenotype of CIK cells was analyzed by the flow cytometry. Bladder transitional cell carcinoma T24 cells were mixed with CIK cells to detect the mortality of the cancer cells with CCK-8 method in vitro. Results With the cultured time of CIK cells prolong,both the number and killing activity increased. the ratio of CIK cells and T24 cells were 1:2. 5,1:5,1:10 and 1:20 when the kill rate of T24 cells by CIK cells were(18. 98 ± 2. 61)%, (35. 59 ± 4. 74)%,(45. 34 ± 5. 03)% and(69. 24 ± 5. 19)%. Conclusion The CIK cells of the cord blood is proved to be with strong anti-cancer activity against T24 cells in vitro.

Objective To evaluate the inhibiting effect of selective and non-selective cyclo-oxygenase-2(COX-2) inhibitor on growth of colon cancer cell lines in vitro and its signal transduction pathway. Methods The proliferation of colon cancer cells was determined by MTT assay. COX-2, nuclear factor(NF)-?Bp65, extracellular-signal regulated kinase(ERK), phospho ERK(pERK) protein expressed in colon cancer cell lines were analyzed by Western blot. Activator protein(AP)-1 and NF-?B binding activity influenced by non-steroidal anti-inflammatory was detected by electrophoretic mobility sift assay. Results Aspirin and celecoxib could inhibit the proliferation of HT-29, SW480 and LS174-T cells and showed a dose-dependent manner. Western blot analysis showed that expression of COX-2, pERK, NF-?Bp65 protein in colon cancer cells were down-regulated by celecoxib or aspirin in some degree but not effect in total ERK expression. AP-1 and NF-?B binding activity could be stimulated by 20% fetal calf serum or tumor necrosis factor-?. Both aspirin and celecoxib could inhibit fetal calf serum-induced AP-1 activation in HT-29 and SW480 cells. Celecoxib could also inhibit tumor necrosis factor induced NF-?B binding activity, but aspirin had little effects on SW480 cells. Conclusion NSAIDs are able to potentially inhibit the growth of colon cell lines in vitro and the mechanism may relate to AP-1 and NF-?B signal transduction pathway.

Objective:To investigate the nutritional risk and the incidence of malnutrition in children with recurrent abdominal Henoch Schonlein purpura(HSP), and observe the changes in the incidence of malnutrition after nutritional intervention.Methods:We retrospectively analyzed 90 children diagnosed with recurrent HSP in our hospital from 2017 to 2019.The cases were divided into two groups according to whether nutritional intervention was performed.The cases from January 2017 to May 2018 were chosen as the control group( n=42), and the cases from June 2018 to December 2019 were chosen as the experimental group( n=48). Based on the treatment of the primary disease, the high nutritional risk cases in experimental group were treated with extensively hydrolysed infant formula for nutritional intervention.The children were evaluated for nutritional risk with the STRONGkids nutrition tool.According to the score results, they were divided into high nutritional risk group and medium nutritional risk group.The Z score was used to evaluate malnutrition, and the degree of malnutrition was compared at admission and discharge. Results:Eighty-one children (90.0%) with recurrent abdominal allergic purpura had a high nutritional risk, 9 cases (10.0%)had a medium nutritional risk.There was no significant difference in the incidence of moderate and severe malnutrition between experimental group and control group at admission[39.6% (19/48) vs.40.5% (17/42), P>0.05]. The incidence of moderate and severe malnutrition in control group was higher than that in experimental group at discharge[66.7% (28/42) vs.22.9% (11/48), P<0.05]. Conclusion:In children with recurrent abdominal HSP, due to severe gastrointestinal symptoms and a high incidence of high nutritional risk, nutritional intervention with extensively hydrolysed infant formula can avoid the occurrence or aggravation of iatrogenic malnutrition during hospitalization.

Methods The maximum tolerated dose( MTD)of water and ethanol extracts of Radix millettiae speciosae were measured for the safety evaluation. Results Maximal tolerated dose( MTD)of water extract was higher than 1 000 g·kg-1 ,which equaled to 110 times the dose for human adults(60 kg). MTD of ethanol extract was higher than 1 700 g·kg-1 ,which equaled to 186 times the dose for human adults(60 kg). Conclusion Radix millettiae speciosae does not have obvious toxicity,thus its routine clinical dose is safe and feasible.

Intellectual property protection is an indispensable management measure in the process of knowledge innovation in the National Traditional Chinese Medicine (TCM) Clinical Research Bases. Longhua Hospital, a National TCM Clinical Research Base , has actively participated in studies and construction of TCM intellectual property policy , laws and regulations , and endeavored to enhance international research cooperation and broaden the international repercussions of TCM . Longhua Hospital made great efforts to strengthen management on intel-lectual property management team and measures, advocacy training, property rights formation and transactions. Intellectual property protection played an important role in the construction of the National TCM Clinical Re-search Base in Longhua Hospital .

AIM: To construct HBx eukaryotic expression vector pEYFP-C1-X and eukaryotic expression vector pGL3-P4 driven by P4 promoter of human IGF-II gene and to investigate the effect of HBx on the transcription activity of IGF-II gene P4 promoter. METHODS: HBx gene and P4 promoters were cloned into pEYFP-C1 and pGL3-basic vectors respectively by gene recombination techniques to construct recombinant plasmids pEYFP-C1-X and pGL3-P4. HepG2 cells were transfected with pEYFP-C1-X and the resistant cell clones were selected by G418. Then methylated pGL3-P4 was transiently transfected into the above cell clones, and the transcription activity of P4 promoter was determined by dual-luciferase reporter assay system. RESULTS: (1) Aim fragments HBx gene and P4 promoter that were cloned were 465 bp and 1 246 bp, respectively and the DNA sequences were accordant with GenBank data confirmed by restricted enzyme digestion and sequencing. (2) HepG2-EYFP-X cells that expressed HBx protein were obtained. (3) Luciferase activity of methylated P4 promoter in the HepG2-EYFP-X was more than that of control cell HepG2-EYFP (P<0.01). CONCLUSION: HBx may enhance the transcription activity of the P4 promoter.

Viperin is an endoplasmic reticulum (ER)-associated protein that has been identified as an innate antiviral protein. Viperin expression can be largely upregulated by viruses, interferons, and oligonucleotides such as poly I:C and lipopolysaccharides. Viperin inhibits viral replication by interactiing with host cell proteins and several viral proteins, and disrupting the cell membrane system. It shows a broad-spectrum of antiviral activity. Some viruses have developed activities that counteract the action of viperin during a long- term period of evolution with hosts by impairing viperin expression. In addition to its antiviral effects, viperin has several other biological functions. This article review the basic characteristics of viperin and the state of current research into its antiviral effects, demonstrating the rapid progress that has been made in this field.
Animals ; Host-Pathogen Interactions ; Humans ; Proteins ; genetics ; immunology ; Virus Diseases ; genetics ; immunology ; virology ; Virus Replication ; Viruses ; genetics ; immunology

Objective To analyze the value of serum procalcitonin (PCT) in patients with community acquired pneumonia (CAP),and to evaluate the role of PCT in the therapeutic effect,severity and prognosis.Methods A retrospective analysis of data and laboratory tests of 50 patients with CAP admitted from November 15,2011 to November 15,2012 in GICU was carried out.Patients with infection of other parts of body,surgical treatment and trauma were ruled out.The level of PCT (ng/mL) before and during treatment,and the relationships between PCT and respiratory failure,mechanical ventilation,treatment results were analyzed respectively.Results According to the occurrence of sepsis,50 patients were divided into sepsis group and non-sepsis group.In the non-sepsis group,the PCT level before treatment,the highest and average PCT levels during the treatment were 0.1125 (0.078,0.269),0.1235 (0.078,0.494),and 0.1355 (0.08,0.245) respectively.Correspondingly,the PCT levels in the sepsis group were 8.92 (2.715,16.33),13.53 (6.305,25.625),and 4.26 (2.1415,8.2455),and there were statistically significant differences in three values of PCT between groups (ZIst =-4.743,PIST ＜ 0.05 ; ZMax =-5.783,PMax ＜ 0.05 ; ZMean =-5.644,PMean ＜ 0.05).According to the emergence of respiratory failure during treatment,average PCT level in the patients with respiratory failure was 1.7375 (0.224,5.092),and that in the patients without respiratory failure was 0.081 ng/mL (0.049,0.146),presenting the statistically significant difference between two groups (Z =4.472,P ＜ 0.05).In case of using mechanical ventilation (MV),the average PCT level of the patients with mechanical ventilation was 1.618 ng/mL (0.224,5.092),and that in the patients without MV was 0.086 ng/mL (0.061,0.465),producing a significant difference between the two groups (Z =-3.788,P ＜ 0.05).Grouped according to the outcome of patients,the mean value of PCT level in death group was 7.4585 ng/mL (2.392,16.25),and that in the survival group was 0.1965 ng/mL (0.885,0.618),showing statistically significant difference between two groups (Z =3.857,P ＜ 0.05).The first PCT level in the GICU within 24 h after admission was used to make the receiver operating characteristic curve (ROC),and the area under the curve (AUC) was 0.9867,cutoff point was 1.25 ng/mL.Conclusions In case of CAP,the PCT level in patients with sepsis is significantly higher than that in patients without sepsis,and PCT can distinguish sepsis from pneumonia precisely.In addition,PCT is an important biomarker to judge the severity and outcomes of CAP at early stage.