Lung cancer ranks the top of malignancies that cause cancer-related deaths worldwide.The leaves of Morus alba L are traditional Chinese medicine widely applied in respiratory diseases.Our previous work has demonstrated the anti-lung cancer effect of secondary metabolites of mulberry leaf,but their mechanism of action has still not fully elucidated.We synthesized Moracin N(MAN)-Probe conjugated with alkyne to label lung cancer cells and identified protein targets by chemical proteomic analysis.MAN and its probe exerted similar growth-inhibitory effect on human lung cancer cells.Chemical proteomic results showed that MAN targeted the programmed death ligand 1(PD-L1)checkpoint pathway and T cell receptor(TCR)signaling pathway,indicating its immune-regulatory function.Cell-free surface plasmon resonance(SPR)results showed the direct interaction of MAN with PD-L1 protein.Molecular docking analysis demonstrated that MAN bound to E158 residue of PD-L1 protein.MAN downregulated the expression levels of PD-L1 in a time-and dose-dependent manner and disrupted the PD-L1/programmed death 1(PD-1)binding,including other secondary metabolites of mulberry leaves Guangsangon E(GSE)and Chalcomoracin(CMR).Human peripheral blood mononuclear cells(PBMCs)co-cultured with MAN-treated A549 cells,resulting in the increase of CD8+GZMB+T cells and the decrease of CD8+PD-1+T cells.It suggested that MAN exerts anti-cancer effect through blocking the PD-L1/PD-1 signaling.In vivo,MAN combined with anti-PD-1 antibody significantly inhibited lung cancer development and metastasis,indicating their synergistic effect.Taken together,secondary metabolites of mulberry leaves target the PD-L1/PD-1 signaling,enhance T cell-mediated immunity and inhibit the tumorigenesis of lung cancer.Their modulatory effect on tumor microenvironment makes them able to enhance the therapeutic efficacy of immune checkpoint inhibitors in lung cancer.

Parkin, an E3 ubiquitin ligase, plays a role in maintaining mitochondrial homeostasis through targeting damaged mitochondria for mitophagy. Accumulating evidence suggests that the acetylation modification of the key mitophagy machinery influences mitophagy level, but the underlying mechanism is poorly understood. Here, our study demonstrated that inhibition of histone deacetylase (HDAC) by treatment of HDACis activates mitophagy through mediating Parkin acetylation, leading to inhibition of cervical cancer cell proliferation. Bioinformatics analysis shows that Parkin expression is inversely correlated with HDAC2 expression in human cervical cancer, indicating the low acetylation level of Parkin. Using mass spectrometry, Parkin is identified to interact with two upstream molecules, acetylase acetyl-CoA acetyltransferase 1 (ACAT1) and deacetylase HDAC2. Under treatment of suberoylanilide hydroxamic acid (SAHA), Parkin is acetylated at lysine residues 129, 220 and 349, located in different domains of Parkin protein. In in vitro experiments, combined mutation of Parkin largely attenuate the interaction of Parkin with PTEN induced putative kinase 1 (PINK1) and the function of Parkin in mitophagy induction and tumor suppression. In tumor xenografts, the expression of mutant Parkin impairs the tumor suppressive effect of Parkin and decreases the anticancer activity of SAHA. Our results reveal an acetylation-dependent regulatory mechanism governing Parkin in mitophagy and cervical carcinogenesis, which offers a new mitophagy modulation strategy for cancer therapy.

Objective: To evaluate the anti-tumor activity of mouse multi-subtype heat shock protein/peptide (mHSP/P) vaccine in combination with a programmed death ligand 1 (PD-L1) inhibitor in mouse sarcoma. Methods: Immunohistochemical staining and en-zyme-linked immunosorbent assay (Elisa) was used to quantitatively identify the expression of heat shock proteins (HSP70, HSP90, Grp94) in the sarcoma cell line MCA207. From the protein suspension prepared, mHSP/P and Grp94/peptide (Grp94/P) sarcoma vac-cines were isolated using chromatography and were identified by Western blot (WB). Flow cytometry was used to determine their cy-totoxic effects. The levels of interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) produced upon mHSP/P and Grp94/P stimulation were measured by Elisa. The effect of sarcoma vaccines on the growth and survival of sarcoma was evaluated in mice. The expression of PD-L1 on the surface of MCA207 sarcoma cells was evaluated by immunofluorescent staining. The effect of IFN-γ treatment on the expression of PD-L1 was determined by WB. Animal experiments explored the effects of PD-L1 inhibitor in combination with mHSP/P treatment on tumors. Results: Tumor tissue carries a variety of HSP subtypes (HSP70, HSP90, Grp94). We successfully isolated sarco-ma tissue-derived mHSP/P and Grp94/P tumor vaccines, which were identified by WB; flow cytometry analysis demonstrated their cy-totoxicity. The levels of IFN-γ and TNF-α cytokines upon mHSP/P stimulation were significantly higher than that observed upon Grp94/P stimulation (P<0.05). The expression of PD-L1 on the surface of sarcoma cells increased with IFN-γ treatment. Animal experiments demonstrated that PD-L1 inhibitor in combination with mHSP/P significantly increased the immune response against tumor (P<0.05). Conclusions: Tumor-derived mHSP/P and Grp94/P can be used as tumor vaccines in animal models. The mHSP/P can elicit a stronger anti-tumor immune response than Grp94/P. IFN-γ stimulates the expression of PD-L1 in sarcoma cells, which results in immune eva-sion. The PD-L1 inhibitor in combination with mHSP/P increased the anti-tumor effect in the tumor microenvironment.

In order to investigate the molecular evolution and spatio-temporal migration of Getah viruses (GETV) isolated around the world,the nucleotide and deduced amino acid sequence of GETVs were analyzed and phylogenetic trees were constructed by using informatics software including ClustalX1.83,MegaAlign,GeneDOC and Mega6.0.The Bayesian Stochastic Search Variable Selection (BSSVS) program in the BEAST v 1.8.1 software package was used to analyze the spatial dynamics of the Getah virus.Results showed that the full-length of Getah virus E2 gene consists of 1 266 nueleotides,encoding 422 amino acids.And the homology of nucleotide and amino acid were 94.5％ 100％ and 96.4％ 100％ respectively.The molecular evolution analysis revealed that there were no species and geographical distribution difference existing among GETV host animals (e.g.horses and pigs) and vectors (e.g.mosquitoes).Bioinformatics analysis showed that GETV originated in Malaysia,then it was spread to Japan,China,South Korea,Mongolia,Russia,etc.GETV E2 gene was relatively stable since GETV was first isolated in 1955.The differences of species and geographical distribution did not exist among GETV host animals and vectors,and the virus has spread from tropical regions to Eurasian continent.Thus,strengthening the detection and monitoring of GETV and its infections in humans and livestock is critical.

Objective To construct lentiviral vectors containing peptide P1-GFP fusion genes.Umbilical cord derived mesenchymal stem cells were infected with lentivirus carrying peptide P1 and GFP fusion genes.To inject the targeted umbilical cord derived mesenchymal stem cells into mice and to detect GFP expression in the spleen.Methods Umbilical cord derived mesenchymal stem cells were cultured with adhered tissues of umbilical cord smaller than 1 mm3 . Lentiviral vector containing P1-GFP fusion genes with engineering technology was constructed and infected the umbilical cord derive mesenchymal stem cells.Targeted umbilical cord derived mesenchymal stem cells were intravenously injected in the mouse tail vein and after 18 hours GFP expression was detected with immunohistochamical staining of the spleen tissues.Results Harvested umbilical cord derived mesenchymal stem cells grew well in culture medium. Green fluorescence on umbilical cord derived mesenchymal stem cells were observed under fluorescence microscope at 18 hours after infected with lentivirus.Green fluorescence intensity of umbilical cord derived mesenchymal stem cells was increasing over time and reached a peak at 72 hours.Umbilical cord derived mesenchymal stem cells highly expressed CD105 (90.0%)/CD44 (98%) and CD73 (85.0%)/CD90 (98.5%) molecules.GFP expression was detected in the spleen after intravenous injection of targeted umbilical cord derived mesenchymal stem cells in the mice 18 hours later.GFP expressing cells intimately contacted with lymphocytes.Conclusions Targeted umbilical cord derived mesenchymal stem cells contain P1-GFP fusion genes are constructed.Targeted umbilical cord derived mesenchymal stem cells can be targeted to mouse spleen and intimately contact with lymphocytes after intravenous injection.Our results lay the groundwork for further studies.

Objective To compare blood pressure control in community hypertensive patients with different management methods.Methods Two neighborhood committees in a community of Pudong were selected as study area using cluster sampling method.A total of 5 166 residents aged ≥35 y were screened for blood pressure; the subjects with high blood pressure and had antihypertensive medication in last 6 months were included,and patients with secondary hypertension was excluded.The patients who entered community hypertension management program and got medication from community were included in community group; those who did not enter in community management program and/or not get medication from community were included in non-community group.Self-designed questionnaire was used for investigation.The medication compliance,awareness of hypertension risk factors and high blood pressure control were compared between two groups.Results Among 5 166 residents 4 763 were surveyed for hypertension with a response rate of 92.2％ and hypertension prevalence rate of 23.2％ (1 105/4 763).Among 1 012 patients with drug treatment for more than 6 months,there were 878 cases in community (86.8％) and 134 cases (13.2％) in non-community group.There were no significant differences in gender,age,education,working condition between community group and non-community group (P ＞ 0.05).44.3％ (389/878) patients in community group had a history of high blood pressure ＞ 10 y and that was 56.7％ (76/134) in non-community group (P =0.011) ; 28.6％ (251/878) patients in community group were at high risk for risk stratification and that in non-community group was 47.8％ (64/134) (P ＜0.001).The awareness of hypertension risk factors in community group and non-community group was 83.9％ and 95.5％,respectively (P ＜ 0.001).The medication compliance and blood pressure control rates in two groups were 93.2 ％ and 84.3 ％ (P ＜ 0.001),68.6％ and 51.5 ％,respectively (P ＜ 0.001).Conclusion The outcomes of hypertension management in terms of medication compliance and blood pressure control in community group are better than those in non-community group.

Objective To investigate the role and clinical significance of RANTES in endometriosis (EM). Methods The serum level of RANTES was examined by ELISA in 50 patients with endometriosis (EM group), 32 patients with benign ovarian neoplasms (disease control group) and 30 normal control women (normal control group). The level of RANTES in peritoneal fluid was examined in EM group and disease control group. Results The serum level of RANTES was significantly higher in EM group (108.73±60.69) ng/L than that of disease control group (31.26±20.33) ng/L and normal control group (29.77 ± 11.58) ng/L (P＜0.05). The level of RANTES in peritoneal fluid was significantly higher in EM group (726.31 ± 259.83) ng/L than that of disease control group (116.19 ± 81.64) ng/L (P＜0.05). The levels of RANTES in serum and peritoneal fluid in EM group were positively correlated with clinical stage respectively (rs=0.501 and 0.562,P＜0.01). The level of RANTES in peritoneal fluid in EM group was positively correlated with dysmenorrhea score (rs=0.527,P＜0.01). The serum level of RANTES was positively correlated with the level of RANTES in peritoneal fluid in EM group (rs=0.363, P＜0.05). The levels of RANTES in serum and peritoneal fluid were positively correlated with inflammatory response degree in endometriotic tissues in EM group (rs=0.326 and 0.391,P＜0.05 or P＜0.01).Conclusion Detection of the serum level of RANTES by ELISA may be one of parameters for diagnosis of endometriosis.

This article discussed the theoretical basis,therapeutic thoughts and concrete methods for treating ankylosing spondylitis on Governor vessel from the aspects of the clinical symptoms of ankylosing spondylitis,the circulation of Governor vessel,traditional Chinese medicine basic theory and clinical application.Moxibustion on Governor vessel can effective improve the stiffness and pain of ankylosing spondylitis.

Objective To evaluate the clinical efficacy and adverse effect of Traditional Chinese Medicine and acupuncture in treatment of ankylosing spondylitis. Methods 46 patients of ankylosing spondylitis with informed consent were randomly divided into observation group and control group by number table,each of 23 cases.The control group received traditional Chinese medicine,the observation group was treated with acupuncture and traditional Chinese medicine,the two groups were treated for 1 month,the clinical effect were observed. Results In the observation group,excellent in 18 cases(78.3%),effective in 3 cases(13.0%),ineffective in 2 cases(8.7%),the total effective rate was 91.3%,it was significantly higher than that of control group the 11 cases(47.8%),8(34.8%),4 cases (17.4%)and 82.6%,the difference between the groups was statistically significant(x2=4.572,P＜0.05).The patients of observation group had a rash in 1 case,1 case of pain;in the control group,1 case of rash,1 case of itching;all of the symptoms were relatively minor,did not affect the treatment.There were no significant adverse reactions in the patients of two groups. Conclusion Traditional Chinese medicine and acupuncture in treatment of ankylosing spondylitis had satisfactory effects and no significant adverse reactions,it was worthy to be recommended in the clinical application.