AIM To study the secondary metabolites from endophytic fungi of Taxus wallichiana var.chinensis (Pilger) Florin and their biological activities.METHODS Endophytes were isolated and purified by tissue block and scribing method,the obtained strains were identified by molecular method.Macroporous resin column and silica gel column were used for separation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The inhibition of phosphatidylinositol 3-kinase,phosphodiesterase and fusarium oxysporum activities were determined by chemiluminescence kinase assay kit method,resazurin cell viability assay and enzyme linked immunosorbent assay.RESULTS Eight endophytic fungi were isolated and identified as Trichoderma,Aspergillus,Gongronella,Phoma,Penicillium.Strain ZGG-5 had strong PI3K inhibitory activity with IC50 value of (0.17±0.05)μg/mL,and was identified as Gongronella butleri.Thirteen compounds were isolated from ZGG-5,and compound 13 had strong activities of inhibiting PI3K at concentration of 1 mg/mL,and the inhibitory rate was (97.8±0.51)％.CONCLUSION T.wallichiana var.chinensis contains endophytic fungi with strong activities of inhibiting PI3K.Compounds 1-12 are first isolated from G.butleri.

AIM To study the secondary metabolites from endophytic fungi of Taxus wallichiana var.chinensis (Pilger) Florin and their biological activities.METHODS Endophytes were isolated and purified by tissue block and scribing method,the obtained strains were identified by molecular method.Macroporous resin column and silica gel column were used for separation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The inhibition of phosphatidylinositol 3-kinase,phosphodiesterase and fusarium oxysporum activities were determined by chemiluminescence kinase assay kit method,resazurin cell viability assay and enzyme linked immunosorbent assay.RESULTS Eight endophytic fungi were isolated and identified as Trichoderma,Aspergillus,Gongronella,Phoma,Penicillium.Strain ZGG-5 had strong PI3K inhibitory activity with IC50 value of (0.17±0.05)μg/mL,and was identified as Gongronella butleri.Thirteen compounds were isolated from ZGG-5,and compound 13 had strong activities of inhibiting PI3K at concentration of 1 mg/mL,and the inhibitory rate was (97.8±0.51)％.CONCLUSION T.wallichiana var.chinensis contains endophytic fungi with strong activities of inhibiting PI3K.Compounds 1-12 are first isolated from G.butleri.

Objective:To explore the optimal time point for combining chemotherapy and immunotherapy and provide an experimen-tal basis for immunotherapy intervention in clinical. Methods:Twenty-three lung cancer patients who completed five chemotherapy cycles between November 2015 and December 2016 in the First Affiliated Hospital of Xinxiang Medical University were enrolled in this study. Numbers of T lymphocyte subsets, B lymphocytes, and NK lymphocytes in peripheral blood were counted. Expression levels of T lymphocyte co-suppression molecule and cytokines in the peripheral blood mononuclear cell were detected using flow cytometry to analyze the dynamic changes of such indicators from one cycle to five cycles of chemotherapy. Results:Significant decreases in the lev-els of CD8+T lymphocytes, CD19+B lymphocytes, and CD16+CD56+NK cells and an increase in CD4+T lymphocytes were observed in the course of multi-cycle chemotherapy for patients with lung cancer. Differences were statistically significant (P<0.05). The co-suppres-sion molecular expression of PD-1, CTLA-4, and CCR-4 with T lymphocytes was downregulated, and the differences were significant (P<0.05). Conclusion:Profiling the dynamic changes of lymphocyte subsets and the expression of T lymphocyte co-suppression molecule are significant in multiple chemotherapy cycles for patients with lung cancer. In the later stage, the combined application of PD-1, PD-L1, CTLA-4, or CCR-4 antibody may exert good therapeutic effects for patients with a high expression level of related immune check-points.

Objective To construct small RNA deletion and overexpression strains with a length of less than 100 nt in Yersinia pestis.Methods Deletion mutants of the target sRNAs were constructed by increasing the length of homologous regions.Meanwhile, the high copy plasmid pBAD/HisA was modified into an inducible transcriptional vector as an sRNA-overexpression plasmid by using QuikChange lightning site-directed mutagenesis kit .The presence , size, and transcription-al initiation sites of the indicated sRNA were predicted by transcriptome sequencing , primer extension , and previous stud-ies.The full-length DNA fragments of target sRNAs were transformed into the transcriptional vector .The overexpressing strains of sRNAs were identified by Northern Blot .Results and Conclusion Four sRNAs deletion mutants of sR01, sR02, sR03 and HmsA and three sRNAs overexpression mutants MicF , HmsA and CpxQ were successfully constructed .A method of construction of sRNA deficient and overexpressing strains of Y.pestis has been quickly and efficiently established by λ-Red homologous recombination technology and QuikChange ? lightning site-directed mutagenesis kit.

Objective To investigate the antitumor effects of IL-18 gene transfected T cells on pancreatic cancer cell SW-1990.Methods Construction of IL-18 contained recombinant lentivirus using PCR method and packaged at HEK293T cell, then transfected the human T cells and evaluated the antitumor effects when cocultured with SW-1990.Results The IL-18 gene contained recombinant lentivirus was successfully constructed and packaged, and transfected the T cells, the LDH secretion and IL-2 and IFN-γ content all increased significantly (P<0.01) when cocultured with SW-1990 cell.Conclusions T cells transfected with IL-18 possessed more potent antitumor effects to pancreatic cancer cell SW-1990 as compared to the regular T cells.

OBJECTIVETo comprehensive evaluate the rationality of Acquired Immune Deficiency Syndrome (AIDS) expenditure through the analysis of AIDS expenditure circumstances based on the application of National AIDS Spending Assessment in Dehong prefecture in 2010.

METHODSDemographic and economic indicators of Mang City, Yingjiang, Longchuan, Ruili City, Lianghe and Wanting zone in Dehong prefecture of Yunnan province were collected from the reports issued by Dehong Statistical Bureau of 2010, and HIV/AIDS epidemic indicators were collected from the annual report of national AIDS prevention and control data.NASA method was used to analyze the actual spending and demand index was used to calculate the demand of AIDS funding in these counties (cities). The correlations between HIV/AIDS expenditures and demographic, economic and HIV/AIDS epidemic index were analyzed, respectively, as well as the correlation between the expenditures of AIDS prevention and control and proportion of major transmission approaches.

RESULTSIn 2010, the actual expenditures on HIV/AIDS in the 5 counties (cities) of Dehong prefecture was ¥28 752 772, the population was 1 211 400, and Gross Domestic Product (GDP) was ¥11 693. The intervention expenditures in injection drug users, sexually transmitted people, and prevention of mother to child transmission in Ruili county were ¥130 345, ¥71 484 and ¥164 100, the proportions of HIV transmission in these groups were 23.9% (49/205), 73.7% (151/205), and 0.5% (1/205), respectively; and in Lianghe county was ¥141 665, ¥257 142, and ¥99 961, and the proportions of HIV transmission were 17.1% (6/35), 80.0% (28/35) and 0.0% (0/35), respectively. The intervention expenditures in Ruili and Lianghe counties were positive related to the proportion of corresponding people with HIV infection (both r values were 0.99, all P values < 0.05). In the other 4 counties (cities) and the development zone, no correlations. The expenditures of government departments, international projects, domestic research programs and personal and family pays (r value were 0.94, 0.83, 0.99 and 0.88, respectively, all P values < 0.05) were positive related to the number.Except the personal and family pays, other sources of expenditures were all positive related to the number of reported HIV/AIDS cases (r values were 0.94, 0.89 and 0.81, respectively, all P values < 0.05).

CONCLUSIONPopulation and HIV/AIDS epidemic index were considered as factors in AIDS spending. The expenditures corresponded with the demand of different areas basically.

Acquired Immunodeficiency Syndrome ; economics ; epidemiology ; prevention & control ; China ; epidemiology ; Health Expenditures ; Humans

OBJECTIVETo calculate the actual expenditures in a county of Dehong prefecture, Yunnan province, China by using the method of National AIDS Spending Assessment (NASA) in 2010.

METHODSData were collected through NASA data collection form based on adapted NASA classification in the county of Dehong prefecture from October to December, 2011, and complemented by semi-structured interview with 16 well trained programmatic and financial representatives in 8 spending units. Data were entered in Resource Tracking Software (RTS) V 2009.3.0, and SPSS 13.0 was used for data processing and analysis.

RESULTSThe NASA estimations showed that the county spent a total of ¥16 235 954 on HIV/AIDS in 2010. Public funds constituted 96.3% of the total expenditure (¥15 630 937), followed by Global Fund which accounted for 3.0% (¥484 585) and private sources which accounted for 0.7% (¥120 432). Findings based on NASA categories showed that AIDS spendings were mainly on 4 areas, and expenditure on Care & Treatment was ¥12 401 382 (76.4% of total expenditure), followed by Prevention which accounted for 14.3% (¥2 325 707), Program Management & Administration which accounted for 7.8% (¥1 268 523) and human resources which accounted for 1.5% (¥240 342). The most beneficial population group was People Living with HIV (PLHIV), accounting for 84.7% of total expenditure. (¥13 753 428), followed by 4.8% for high risk population, including female sex workers and their partners (¥297 333), injection drug users and their partners (¥293 143), men having sex with men and their partners (¥185 136) and 1.5% (¥241 429) for the general population.

CONCLUSIONThe local funds for HIV/AIDS in this county was insufficient. The local government should increase corresponding funds based on central government funding. Care and treatment was the first spending priority in the county and the investment of prevention services needs to be increased. Prevention and treatment and care should be combined to ensure the effectiveness of comprehensive prevention and treatment of AIDS.

Acquired Immunodeficiency Syndrome ; economics ; prevention & control ; China ; Financing, Government ; economics ; Health Expenditures ; Humans ; Outcome and Process Assessment (Health Care) ; Surveys and Questionnaires

OBJECTIVETo explore a new method for finding more HIV/AIDS.

METHODSIn September 2009, newly reported HIV-infected individuals from May to August, 2009 in Dehong prefecture were asked to participate in a survey which requested demographic characteristics, history of high-risk behaviors and contact information of individuals with whom they had high risky contacts. People with risky contacts with HIV-infected cases (index cases) were also approached to participate in this survey and HIV testing was provided.

RESULTSA total of 342 HIV-infected individuals were newly reported and served as index cases from May to August, 2009. Among them, 47.1% (161/342) were transmitted by regular sexual partners. Through three-round surveys, 218 contacts were traced and among them, 84.9% (185/218) were traced by regular heterosexual partners. HIV positive rate of the tested was 34.7% (60/173) in the first-round tracing and 12.5% (1/8) in the second one. Among the 560 individuals (index cases and their high-risk contacts), the proportions of having regular heterosexual partners, non-regular and non-commercial sexual partners, commercial sexual heterosexual partners and men having sex men were 87.9% (492/560), 18.9% (106/560), 22.3% (125/560) and 0.3% (1/318), respectively, while the proportion of having never used condoms when having sex with the above four types sexual partners were 73.8% (363/492), 72.6% (77/106), 63.2% (79/125) and 0.0% (0/1), respectively.

CONCLUSIONAs an epidemiological method for HIV/AIDS finding, contact tracing identified a large number of HIV infectors who were traced by newly reported HIV-infected individuals.

Acquired Immunodeficiency Syndrome ; epidemiology ; Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Contact Tracing ; methods ; Female ; HIV Infections ; epidemiology ; Humans ; Infant ; Infant, Newborn ; Male ; Risk Factors ; Surveys and Questionnaires ; Young Adult

OBJECTIVETo study the chemical constituents of the root of Hemerocallis fulva.

METHODCompounds were isolated by repeated silica gel, Sephadex LH-20, MCI gel, etc. column chromatography and their structures were determined by spectral analyses and physicochemical properties.

RESULTFrom the n-butanol fraction of the EtOH extract of the roots of H. fulva, 9 glycosides were isolated and identified as sweroside (1), laganin (2), picraquassioside C (3), puerarin (4), 3'-methoxypuerarin (5), 7-hydro xylnaphthalide-O-beta-D-glucopyranoside (6), orcinol-3-O-beta-glucopyranoside (7), HN saponin F (8), hederagenin-3-0-beta D-glucopyranosyl-(1-3)-alpha-L-arabinopyranoside-28-O-beta-D-glucopyranosyl ester (9).

CONCLUSIONCompounds 1-9 were isolated from the genus Hemerocallis for the first time.

Glucosides ; chemistry ; isolation & purification ; Hemerocallis ; chemistry ; Iridoid Glucosides ; Iridoids ; chemistry ; isolation & purification ; Isoflavones ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Plant Roots ; chemistry ; Saponins ; chemistry ; isolation & purification

OBJECTIVEIn order to explore a new method to perform the medial canthoplasty applied with upper eyelid fold formation in a single procedure.

METHODSAccording to the principles to correct the deformity of the medial canthus, a new method was designed to fetch up the skin defect of the medial side of upper eyelid, minimize the skin scarring around the medial canthus, and form upper eyelid fold.

RESULTSThe 86 cases, which mild or moderate epicanthus without upper eyelid fold, were treated with this technique and long-term follow-ups above 6 months were made. The 67 cases were given the follow-ups. The appearances of upper eyelid fold and medial canthus were evaluated. The appearances of eyelid fold were paralleled type. The epicanthus was corrected completely or in major.

CONCLUSIONSThis new canthoplastic method with upper eyelid fold formation may be suitable to all the medial epicanthus except inverse epicanthus. All of the patients were satisfied with the final results.

Adolescent ; Adult ; Blepharoplasty ; methods ; Eyelids ; surgery ; Humans ; Middle Aged ; Young Adult