Evasion of apoptosis is a hallmark of cancer, attributed in part to overexpression of the anti-apoptotic protein B-cell lymphoma 2 (Bcl-2). In a variety of cancer types, including lymphoma, Bcl-2 is overexpressed. Therapeutic targeting of Bcl-2 has demonstrated efficacy in the clinic and is the subject of extensive clinical testing in combination with chemotherapy. Therefore, the development of co-delivery systems for Bcl-2 targeting agents, such as small interfering RNA (siRNA), and chemotherapeutics, such as doxorubicin (DOX), holds promise for enabling combination cancer therapies. Lipid nanoparticles (LNPs) are a clinically advanced nucleic acid delivery system with a compact structure suitable for siRNA encapsulation and delivery. Inspired by ongoing clinical trials of albumin-hitchhiking doxorubicin prodrugs, here we developed a DOX-siRNA co-delivery strategy via conjugation of doxorubicin to the surface of siRNA-loaded LNPs. Our optimized LNPs enabled potent knockdown of Bcl-2 and efficient delivery of DOX into the nucleus of Burkitts' lymphoma (Raji) cells, leading to effective inhibition of tumor growth in a mouse model of lymphoma. Based on these results, our LNPs may provide a platform for the co-delivery of various nucleic acids and DOX for the development of new combination cancer therapies.

Objective: To investigate the function of anti-PD-1 (scFv)/IL-15/IL-15Rα-sushi fusion protein (PD-S15) to specifically bind to PD-1 in vitro and to explore its effect on NK/T cell proliferation. Methods: The human anti-PD-1 (scFv) gene sequence and human IL-15/IL-15Rα-sushi fusion gene sequence were synthesized chemically. The recombinant expression plasmid pUC57-PD-S15 was constructed by enzyme digestion and ligation of the two target genes, and then transiently transfected into HEK293T cells by lipofectamineTM 2000. The supernatants of cell culture medium were acquired, and the expression of PD-S15 fusion protein in cell culture supernatants was detected by Wb assay. PBMCs and TILs were cultured in mediums with different proportion of PD-S15/X-VIVOTM15, respectively. Then, the capacity of PD-S15 fusion protein to bind to PD-1 in vitro and its effect on the proliferation of PBMCs and the proportion of CD3+CD8+, CD3+CD4+ and CD3-CD56+ subsets were detected by flow cytometry. The effect of PD-S15 fusion protein on the proliferation of TILs was detected by cytometry. Results: The successful construction of pUC57-PD-S15 eukaryotic expression plasmid was confirmed by double enzyme digestion and sequencing, and then successfully transfected into HEK293T cells. The relative molecular weight of the target protein was approximately 55 000, and was in line with expectations. PD-S15 fusion protein could specifically combine with PD-1 in vitro (P<0.05) and stimulate NK/T cell proliferation (P<0.05). Compared with classical TILs culture method, the efficiency of activation and amplification of T cells in vitro by PD-S15 culturemethodwasbetter (P<0.01). Conclusion: PD-S15 fusion protein can specifically target PD-1 and rapidly expand NK/T cells in vitro, which lays a foundation for the selective expansion of CD8+PD-1+ antigen-specific T lymphocytes from tumor tissues and even peripheral blood.

A qualitative analytical method of liquid chromatography coupled with mass spectrometry(LC-MSn)was developed for the identification of main constituents in Chrysanthemum morifolium ‘Fubaiju’. High-performance liquid chromatography(HPLC)was developed for the quantification of five active components, including chlorogenic acid(1), luteolin-7-O-β-D-glucopyranoside(2), luteolin-7-O-β-D-glucopyranuronide(3), 3, 5-Di-caffeoylquinic acid(4), and apigenin-7-O-β-D-glucopyranoside(5). A total of 22 compounds, including 13 flavonoids and 9 phenolic acids, were identified based on their retention behaviors, UV profiles and MS fragment information. Furthermore, a validation method with good linearity(r> 0. 999 9), precision, stability, repeatability and recovery was successfully applied for simultaneous determination of five major components in 10 batches of C. morifolium ‘Fubaiju’ by HPLC-UV method. The established method was proved to be a validation strategy for the quality evaluation of C. morifolium ‘Fubaiju’.

Objective: To investigate the clinical value of combined detection of serum miR-378 and miR-21 in gastric cancer (GC). Methods: Eighty-seven patients with GC and 78 patients with colorectal cancer(CRC) from National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences were selected, 83 individuals undergoing healthy physical examination were selected as the healthy controls. The levels of serum miR-378 and miR-21 were detected by quantitative real-time PCR (RT-qPCR) (result data were transformed as log2 for analysis). Results: Relative expression levels of miR-378 in the serum were -1.24, -3.25 and -2.73 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-378 were significantly decreased in GC and CRC patients (both P<0.05). Relative expression levels of miR-21 in the serum were 0.11, 2.34 and 2.47 in healthy controls, GC and CRC patients, respectively. Compared with the healthy controls, the levels of serum miR-21 were significantly up-regulated in GC and CRC patients (both P<0.05). Moreover, the serum level of miR-378 in GC patients was inversely associated with tumor clinical stage (P<0.05). However, the level of miR-21 showed no significant differences among patients with different clinical and pathological characteristics (all P>0.05). The area under the receiver operating characteristic curve (AUC), sensitivity and specificity of miRNA-378 to diagnose GC was 0.770, 82.0% and 66.0%, respectively, and were 0.900, 85.0%, and 88.0% of miR-21, respectively. The AUC, sensitivity and specificity of combined detection of serum miR-378 and miR-21 to diagnose GC were 0.930, 92.0% and 87.0%, respectively, while the AUC of combined detection of serum CEA and CA-199 was 0.767, the AUC of combined all of the four factors was 0.946. Conclusion The combined detection of serum miR-378 and miR-21 have a certain effect on diagnosis of GC.

Objective To investigate the clinical significance of HCV antibody S /CO values in active HCV infection diagnosis in cancer patients .Methods 390 cancer patients were enrolled from Cancer Hospital Chinese Academy of Medical Sciences between January 2013 and April 2015.All of the cancer patients had pathological diagnosis , including 240 males and 150 females, aged from 25 to 83 years old. HCV antibody and HCV RNA levels were detected using the Abbott i 2000 immunity analyzer and Roche LC480 real-time fluorescent quantitative PCR machine , respectively.The relationship between HCV antibody S/CO value and RNA level was analyzed in the group of HCC and non-HCC patients.Results There were obvious statistical differences in age (P＝0.004), gender (P＜0.001) and HCV antibody levels (P＜0.001) between the group of HCC and non-HCC patients.There was no statistical difference in distribution of RNA positive rate between the two groups (P＝0.528).Using ROC curve analysis, the best cut-off value to diagnose active HCV infection in cancer patients is 10.0 with sensitivity 97.6%and specificity 81.3%. According to the results of the ROC curve , the cut-off was 11.4 and 10.4 in HCC and non-HCC patients respectively.Conclusion The best cut-off value to diagnose active HCV infection in cancer patients is 10.0, either in HCC or in non-HCC.

Phenylalanine hydroxylase (PAH) is a member of aromatic amino acid hydroxylase (AAAHs) family, and catalyze phenylalanine (Phe) into tyrosine (Tyr). Using immunological and RT-PCR methods to prove the existence of phenylalanine hydroxylase (PAH) gene in the brain of Neanthes japonica in protein and nucleic acid level. Using Western blotting to detect the pah immunogenicity of Neanthes japonica. Making paraffin sections and using immunohistochemical technique to identify the presence and distribution of the phenylalanine hydroxylase gene in the brain of Neanthes japonica. Clone pah gene from the brain of Neanthes japonica by RT-PCR, constructing plasmid and transferring into E. coli to amplification, picking a single homogeneous colony, double digesting then making sequence and comparing homology. Western blotting results showed that the expression of the protein is present in Neanthes japonica brain, immunohistochemistry technique results showed that phenylalanine hydroxylase mainly expressed in abdominal of forebrain, dorsal and sides of midbrain. RT-PCR technique results showed that the phenylalanine hydroxylase exist in the brain of Neanthes japonica and has a high homology with others animals. PAH is present in the lower organisms Neanthes japonica, in protein and nucleic acid level. Which provide the foundation for further study the evolution of aromatic amino acid hydroxylase genes in invertebrate.
Animals ; Blotting, Western ; Brain ; enzymology ; Escherichia coli ; metabolism ; Phenylalanine Hydroxylase ; genetics ; metabolism ; Polychaeta ; enzymology ; genetics

OBJECTIVE:To evaluate clinical efficacy through comparing the change of CT image in infrapatellar fat pad before and after Xiaoding ointment in the treatment of acute patellar bursitis of knee joint. METHODS:73 patients with acute patellar bur-sitis were randomly divided into observation group(39 cases)and control group(34 cases). Observation group was given Xiaoding ointment for local application,qd,7 d as a courses,3 courses in total;control group was given triamcinolone acetonide 30 mg af-ter the extraction of articular cavity effusion,once a week,totally for 3 times. All patients of two groups underwent knee CT exami-nation for observation of the infrapatellar fat pad and articular cavity effusion volume change before and after treatment. Clinical ef-ficacies were compared between 2 groups. RESULTS:CT image alterations of treatment group showed that infrapatellar fat pad den-sity were decreased,anteroposterior diameter,vertical diameter,internal to external diameter were significantly reduced. The total effective rate of treatment group was 92.31%,which was better than that of control group(88.24%),with statistical significance (P<0.05). CONCLUSIONS:Xiaoding ointment demonstrate markedly curative effects in the treatment of acute patellar bursitis, and CT image is an effective method for diagnosis of infrapatellar fat pad.

Objective To analyze the correlation between the polymorphism of PTPN22 gene (R620W and R263Q sites)and pulmonary tuberculosis(TB)in Chinese Han population and to investigate the environmental factors associated with pulmonary TB. Methods A case-control study was conducted on 235 patients with pulmonary TB and 251 healthy subjects. The single nucleotide polymorphisms(SNP)of PTPN22 gene at R620W and R263Q sites were detected by the assay of polymerase chain reaction and re-striction fragment length polymorphism(PCR-RFLP). A questionnaire was designed to gather information about tuberculosis-associated environmental factors. Univariate and multivariate logistic analysis were con-ducted. Results Genotype frequencies of PTPN22 R620W(C1858T)SNP were 233(99. 15% ,CC),2 (0. 85% ,CT),0(0% ,TT)in patients with pulmonary TB and 240(95. 62% ,CC),11(4. 38% ,CT), 0(0% ,TT)in healthy subjects. There was a difference with the distribution of PTPN22 C1858T allele be-tween patients with pulmonary TB and healthy subjects[0. 43% vs 2. 19% ,P = 0. 01,odds ratio(OR)=0. 19,95% confidence interval(CI)= 0. 07-0. 35]. Genotype frequencies of PTPN22 R263Q(G788A) were 218(92. 77% ,GG),17(7. 31% ,GA),0(0% ,AA)in patients with pulmonary TB and 248 (98. 71% ,GG),3(1. 29% ,GA),0(0% ,AA)in healthy subjects. The frequencies of G788A allele in patients with pulmonary TB were higher than those in healthy subjects(3. 62% vs 0. 60% ,P﹤0. 01,OR=6. 03,95% CI=2. 12-18. 38). Conclusion The results of this study suggested that the R263Q GG geno-type of PTPN22 gene was associated with the susceptibility to TB in Chinese Han population.

BACKGROUND:Increasing autologous stem cellmobilization is conceived to achieve effectively repair of cardiac ischemic injury. Therefore, it is important to seek a specific and effective mobilization agent. OBJECTIVE:To observe the effects of hypoxia-inducible factor-1α(HIF-1α) on bone marrow mesenchymal stem cellmobilization in myocardial infarction. METHODS:Left anterior descending artery was ligated to establish a rat model of acute myocardial infarction in 90 outbreeding Sprague-Dawley rats, and then the models were randomly divided into three groups. In HIF-1α-antisense oligonucleotide (ASODN) group, HIF-1α-ASODN was infused into the tail vein to restrain the expression of HIF-1αin infarcted ischemic tissue. In HIF-1α-missense oligonucleotide (MSODN) group or control group, an equal volume of HIF-1α-MSODN or saline was injected. RESULTS AND CONCLUSION:After 30 hours and 7 days of modeling, the number of bone marrow mesenchymal stem cells and expression of vascular endothelial growth factor in the peripheral blood of the control group were similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. After 7 days of modeling, the expressions of HIF-1αprotein, vascular endothelial growth factor protein and mRNA in the ischemic myocardial tissues of the control group were similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. After 7, 14 and 28 days of modeling, the capil ary density in the ischemic myocardial tissues of the control group was similar to the HIF-1α-MSODN group, but significantly higher than the HIF-1α-ASODN group. These findings indicate that after acute myocardial infarction, high expression of HIF-1αexhibits a causal relationship with mobilization of bone marrow mesenchymal stem cells, initiating a series of self-healing process of myocardial tissues.

Objective To investigate the incidence of contrast induced nephropathy (CIN) among different patient groups after contrast agent injection.Methods A total of 1243 patients were included in this study (male =694,female =549).The SCr level one week before and 72 hours after the CT examination and the incidence of CIN were recorded and comparison was made among groups according to sex,age,body mass index (BMI),the history of high blood pressure (HBP),diabetes mellitus (DM),chronic kidney disease (CKD),chronic heart failure (CHF),tumor,nephrotoxicity drug (NTD) usage.The frequency,type,dose and injection velocity of the contrast media(CM)were also recorded.Multivariate predictors of CIN were identified by Logistic regression using step-wise selection with entry and exit criteria of P ＜0.10,results were tabulated as odds ratios (OR) with 95％ confidence intervals (CI).Results Among 1243 consecutive patients,the incidence of CIN was 5.5％ (68/1243).Patients with a history of HBP,DM,CHF,CKD or tumor presented with higher incidence of CIN than that of controls (5.9％,51/868 vs.4.5％,17/375).CIN developed in 9 of 203 patients (4.4％,9/203) with CKD and in 59 of 1040 patients (5.7％,59/1040)without CKD.There was no significant difference between the two groups(x2 =0.51,P =0.30).In CKD (-) group,the incidence of CIN was higher in females,patients with DM and patients using LOCM than those of males,DM (-) and using low osmolality contrast medium (IOCM) (P ＜ 0.05),but there was no statistical significance in CKD (+) group.Logistic regression analysis showed that women,age ≥ 75 years,DM,LOCM,NTD,tumor,the time of using CM more than once per month were the most significant predictors of CIN (OR ＞ 1).Conclusion Women,age ≥ 75 years,LOCM,NTD,tumor,and the frequency of using CM more than once per month were more likely to develop CIN.