Objective: To explore the active constituents from the stems of Orostachys malacophyllus in Changbai Mountain. Methods: The ethyl acetate fraction of the methanol extract from the stems of O. malacophyllus was separated by chromatographic methods, structure identification was by means of nuclear magnetic resonance and mass spectra, and the activity was determined by CCK-8 experiment. Results: Five compounds were isolated and identified as 1-ethyl-2-hydroxysuccinate (1), 1-ethyl-2-O-[(3,4-dihydroxybenzoyl) oxy]-succinate (2), 1,4-dimethy-2-O-[(3,4-dihydroxybenzoyl) oxy]-succinate (3), 1,4-dimethy-2-O-[(4-dihydroxybenzoyl) oxy]-succinate (4), and diisobutyl phthalate (5). Conclusion: Compounds 1-5 are isolated from the plants of this genus for the first time. Among them, compound 2 is a new compound named orostachysolic acid, and exhibits cytotoxicity against HeLa cell with IC50 value of 111.5 μmol/L.

AIM: To better reveal the changing process of macula area after photodynamic therapy (PDT), the changes of tomographic imaging on macular fovea optical coherence tomography ( OCT ) scan at different follow- up time points after treating age-related macular degeneration ( AMD) by PDT were investigated.
METHODS: This cohort study included 30 eyes of 26 patients diagnosed as AMD. The dosage of verteporfin was calculated according to manufacturer's instruction. All participants got FFA, ICG, OCT scan and best corrected visual acuity before and followed up for 2wk, 1, 2, 3mo after PDT. The standard five lines combined with 6 lines OCT scan covered key parts of lesion and the scan locations before and after were kept the same. The bilaminar foveal thickness (BFT), outer high reflectivity band thickness ( OHRBT ) , and the total area of intraretinal fluid ( IRF ) and subretinal fluid ( SRF ) were measured at different time points. The relationship between the changes and follow up time was analyzed. Statistical analysis of the data was performed using SPSS for windows version 13. 0.
RESULTS: There months after treatment, an improvement of 2 lines or better on the Snellen chart was achieved on 22 eyes, visual acuity of 3 eyes without significant change, 3 eyes got decreased, 2 cases lost follow up. The average macular thickness of improved 22 eyes before treatment was 722. 5±55. 6μm, 2wk after treatment, 708. 3±45. 3μm, 1mo, 584. 4±49. 3μm, 2mo, 430. 7±50. 2μm and 180. 6 ±36. 3μm at 3mo. The OHRBT before and after treatment respectively were 302. 3 ±50-2μm, 277. 5±42. 3μm, 202. 7±40. 1μm, 180. 6±35. 7μm, 100. 8±22. 9μm. The total area of both IRF and SRF was estimated as 0. 34±0. 12mm2 , 0. 25±0. 07mm2 , 0. 10±0-05mm2 , 0. 08±0. 04mm2 , 0. 05±0. 01mm2 .
CONCLUSION:SRF and retinal edema of patients with AMD were absorbed 1mo after PDT, and the BFT, OHRBT were significantly tend to be thin.

AIMTo discover new drugs which may be applied to diseases of the immune system, hemogenesis system diseases and tumors, several high-throughput drug screening cell models based on JAK-STAT signal pathway have been established.

METHODSFour repeats of STAT DNA binding conserved sequences were synthesized, subcloned into pGL-Luc reporter vector and stably transfected into cell lines in vitro. Cell clones with high copy numbers of STAT binding sites and reporter genes were chosen as high-throughput drug screening cell models. The cell models were tested with known anti-allergic drugs and anti-tumor drugs by determining luciferase activity. The reaction was performed in 96 well micro-plates with a final volume of 50 microL.

RESULTSThe cell models by performing rapid fluorescence assay were shown to be highly sensitive and stable after testing with cytokine and drugs. The modification of the expression plasmid simplified this method and made it more practical. It also provided good linear correlation, wide range of assay, highly sensitive and good reproducibility.

CONCLUSIONThe method can be performed by high-throughput drug screening for effective extraction of Chinese traditional herbs.

Anti-Allergic Agents ; isolation & purification ; pharmacology ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; DNA-Binding Proteins ; genetics ; metabolism ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Janus Kinase 2 ; Jurkat Cells ; metabolism ; Liver Neoplasms ; pathology ; Luciferases ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; STAT3 Transcription Factor ; Signal Transduction ; Trans-Activators ; genetics ; metabolism ; Tumor Cells, Cultured

Squalene synthase (SQS) is a key enzyme in plant terpenoid biosynthetic pathway. This study focused on cloning and analysis of Huperzia serrata SQS (HsSQS1) gene. After searching the transcriptome dataset of H serrata, one unique sequence encoding SQS was discovered. The primers were designed according to the transcript sequence of HsSQS1 from the H. serrata transcriptome dataset. The open reading frame of HsSQS1 was cloned using RT-PCR strategy. The bioinformatic analysis of this gene and its corresponding protein were performed. The cDNA (named as HsSQS1) contains a 1263 bp open reading frame and encodes a predicted protein of 420 amino acids. The GenBank accession number for this gene is JQ004938. HsSQS1 contains two transmembrane regions, without signal peptide. The conserved domain of squalene synthase was presented in HsSQS1. HsSQS1 was more abundant in H. serrata root than in leaf and stem. This study cloned and analyzed squalene synthase gene from H. serrata for the first time. The result will provide a foundation for exploring the mechanism ofterpenoid biosynthesis in H. serrata plants.
Amino Acid Sequence ; Biosynthetic Pathways ; Cloning, Molecular ; DNA, Complementary ; genetics ; Expressed Sequence Tags ; Farnesyl-Diphosphate Farnesyltransferase ; genetics ; isolation & purification ; metabolism ; Genes, Plant ; genetics ; Huperzia ; enzymology ; genetics ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Plant Leaves ; enzymology ; Plant Roots ; enzymology ; Plant Stems ; enzymology ; Plants, Medicinal ; enzymology ; genetics ; Triterpenes ; chemistry

OBJECTIVETo study the prevalence rate of hypertension and the risk factors in Guangxi Hei Yi Zhuang population.

METHODSA total of 1068 people of Hei Yi Zhuang nationality aged 20 and over were surveyed. Blood pressure, height, weight, serum lipid and apolipoprotein levels were measured ,and both body surface areas and body mass index were calculated. Results were compared with those in 933 people of Han nationality living in the same district.

RESULTSThe prevalence rates of hypertension in Hei Yi Zhuang population were significantly higher than that in Han population (25.2% vs. 17.3%, P < 0.001) ,as well as the isolated systolic hypertension (12.5% vs. 3.9%, P < 0.001). The mean levels of systolic pressure and pulse pressure in Hei Yi Zhuang population were significantly higher than those in Han population (125.20 +/- 18.62 vs. 121.88 +/- 15.99 mm Hg, P < 0.001 and 48.64 +/- 14.75 vs. 44.98 +/- 11.12 mm Hg, P <.001; respectively). The prevalence rate of hypertension in Hei Yi Zhuang population was positively correlated with triglyceride, sex and age whereas the prevalence rate of hypertension in Han population was positively correlated with total cholesterol, sex, age, and alcohol consumption. Rates of awareness on hypertension in Hei Yi Zhuang and Han population were 8.6% vs. 21.1% (P <.001), patients with treatment rate of established hypertension were 4.5 vs. 15.5 (P < 0.001) with the control rates as 1.9 vs. 10.6 (P < 0.001).

CONCLUSIONThe prevalence rate of hypertension in Guangxi Hei Yi Zhuang population was significantly higher than that in Han ethnic group. The causes of high prevalence rate of hypertension might be ascribed to special geographical surroundings, unhealthy life style, high sodium intake, low education, and possibly genetic factors. The rates on awareness, treatment and control of hypertension were still under satisfaction.

Adult ; Aged ; Aged, 80 and over ; China ; epidemiology ; ethnology ; Ethnic Groups ; Female ; Health Knowledge, Attitudes, Practice ; Humans ; Hypertension ; epidemiology ; therapy ; Logistic Models ; Male ; Middle Aged ; Multivariate Analysis ; Residence Characteristics ; statistics & numerical data ; Risk Factors ; Young Adult

Purpose To study the clinical features, immunophenotypes and prognostic factors of primary breast diffuse large B-celllymphoma (PBDLBCL). Methods The clinical pathological data of 49 cases of PBDLBCL during January 2006 to December 2016 were retrospectively analysed, and the basic clinical and pathologic data, pathologic types and the immunohistochemical slides by EnVision method for staining were summarized. Results 47 cases were women and 2 cases were men. The age ranged from 24 to 79 year old with the median age of 48 year old. On microscopic observation, tumor cells were large to medium-sized which characterized as diffuse infiltration between the lobules of mammary gland, around the duct, interstitial and fat tissue, some were single file cord pattern. The immunophenotype showed 37 cases were of non-GCB, 12 cases were GCB type. Ki-67 index were greater than 40％. According to Ann Arbor staging, 16 cases were stage I EA, 28 cases were stage Ⅱ EA, 5 cases were stage Ⅳ E. IPI score: 30 cases with 0 ～1 score, 10 cases with 2 score, 9 cases with 3 score. Patients were followed up from 5 to 146 months, The 3-year overall survival (OS) rate was 51.2％ and 5-year OS rate was 36.7％, Single factor analysis showed that there were statistically significant difference in clinical stage, levels of LDH, IPI score, BCL-2 protein expression, and BCL-6 protein expression in 3 and 5 years of OS rate. The multiple factor analysis of Cox regression showed that the increase of IPI risk classification was the independent adverse prognostic factor of primary breast diffuse large B-cell lymphoma. Conclusion The diagnosis of PBDLBCL is confirmed by pathological biopsy and immunohistochemical markers. The immunophenotype was mainly non-GCB type. Comprehensive treatment including surgery, chemotherapy and radiotherapy is appropriate. The prognosis should be comprehensively evaluated by multiple factors. IPI increase risk classification is the independent adverse prognostic factor.

OBJECTIVETo investigate the clinicopathological features of primary gastrointestinal non-Hodgkin's lymphomas (PGI-NHL) and their prognostic values.

METHODSThe clinical and pathological data of 216 patients diagnosed as PGI-NHL from Zhejiang Cancer Hospital were analyzed retrospectively. χ² test, log-liner model analysis, COX proportional hazard regression analysis and Life-table survival analysis were used to analyze the survival status of the patients by SAS 8.2 software, and Log-rank test was performed to couple the overall survival rates with different prognostic factors.

RESULTSTotally, the age of onset was 8 to 89 years with the median age of 56.5 years. Male versus female was 1.27∶1(121∶95). The most frequently involved location was stomach (147 cases, 68.1%), followed by ileocecus (25 cases, 11.6%), large intestine (20 cases, 9.3%), small intestine (17 cases, 7.9%) and multiple GI involvement (5 cases, 2.3%). 182 cases were classified as B cell lymphomas, 22 cases as T cell lymphomas, and 12 cases not classified exactly due to insufficient data. The 3-year and 5-year survival rates of the patients were 69.4% and 53.3%, respectively. Univariate analysis revealed that age＞60 years, ECOG≥2, high LDH level, stage Ⅲ-Ⅳ, IPI≥2, T cell type and intestinal involvement were predictors for poor prognosis. IPI≥2, T cell type and intestinal involvement were independent adverse predictors for prognosis by multiple COX proportional hazard regression analysis. Among different treatment groups, cases received chemotherapy combined with local radiotherapy gained the best survival status.

CONCLUSIONB-cell lymphoma was the main pathological type in PGI-NHL; IPI≥2, T-cell type and intestinal involvement are independent adverse predictors for prognosis; chemotherapy combined with local radiotherapy might be the choice of approach for advanced stage and aggressive PGI-HNL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Gastrointestinal Neoplasms ; drug therapy ; pathology ; Humans ; Lymphoma, Non-Hodgkin ; drug therapy ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Young Adult

OBJECTIVETo figure out the function of C/EBPalpha in the monocytic differentiation of HL60 cells induced by a new steroidal drug NSC67657.

METHODSThe differentiation of HL60 cells was induced by NSC67657, and the cell surface antigen CD14 expression was detected by flow cytometry. The gene and protein expressions of CCAAT enhancer binding protein alpha (C/EBPalpha) before and after the induction of cell differentiation were determined by RT-PCR and Western blot. Eukaryotic expressing vector pDsRed-ICAT was constructed and transfected into HL60 cells, and its expression was verified. The effect of C/EBPalpha overexpression in HL60 cells was assessed by MTT assay, Wright's staining and flow cytometry before and after NSC67657 transfection.

RESULTSHL60 cells could be induced into monocytes by 10 micromol/L ATRA within 5 days, and the coverage of CD14 positive cells reached 93.9% after 5 days of drug treatment. The eukaryotic expressing vector was successfully constructed, and over 90% positive clones were obtained after screening by G418 and electrotransfection. The results of proliferative analysis, chemical staining, ultrastructural observation, and CD11b detection confirmed that HL60 cells could be induced into granulocytic differentiation by overexpression of C/EBPalpha protein. Moreover, in the drug treatment group, transfected cells could not be induced into monocytic differentiation, and their granulocytic differentiation was also inhibited.

CONCLUSIONThe monocytic differentiation of HL60 cells induced by NSC67657 may not be via the regulation by C/EBPalpha protein-mediated signal transduction. However, the overexpression of CEBPalpha may inhibit the process of NSC67657-induced monocytic differentiation in HL60 cells.

CCAAT-Enhancer-Binding Protein-alpha ; genetics ; metabolism ; CD11b Antigen ; metabolism ; Cell Differentiation ; drug effects ; Genetic Vectors ; Granulocytes ; cytology ; HL-60 Cells ; Humans ; Lipopolysaccharide Receptors ; metabolism ; Mesylates ; pharmacology ; Monocytes ; cytology ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Signal Transduction ; Steroids ; pharmacology ; Transfection

This study was purposed to explore the effect of different cytokine combinations on the expansion of the mononuclear cells drived from umbilical cord blood (CB) ex vivo and expression of CXCR4 and CD49d on CD34+ cells after expansion. Human fresh CB mononuclear cells were cultured in serum-free and stroma-free medium containing different combinations of cytokine for 7 days. At day o and 7, the total cells were counted, CD34+ cells and CD34+CXCR4+, CD34+CD49d+ cells were assayed by flow cytometry, and CFU were determined. According to the different combinations of cytokine, experiments were divided into four groups: control, SF group (SCF + FL), SFT group (SCF + FL + TPO) and SFT6 group (SCF + FL + TPO + IL-6). The results showed that the SF (SF group) combination supported only low expansion of total cells, CD34+ cells and CFU. The addition of TPO in SF group restored UCB stem/progenitors expansion to a higher level than that in SF group, while there was no difference between groups SFT and SFT6 (P > 0.05). The cytokine combinations in groups SF, SFT and SFT6 all could upregulate the expression levels of CD49d and CXCR4 on expanded cord blood CD34+ cells, but there were no significant differences between groups SF, SFT and SFT6 (P > 0.05). It is concluded that SCF + FL has no strong synergistic effects on primitive hematopoietic cells. TPO plays an important role in enhancing expansion of umbilical cord blood hematopoietic cells, while IL-6 only shows a neutral effect on it. SCF + FL + TPO combination not only promotes progenitor cells expansion but also upregulates the expression of CD49d and CXCR4 on CD34+ cells from cord blood.
Antigens, CD34 ; biosynthesis ; genetics ; Cytokines ; pharmacology ; Drug Synergism ; Fetal Blood ; cytology ; Humans ; Integrin alpha4 ; biosynthesis ; genetics ; Leukocytes, Mononuclear ; cytology ; Membrane Proteins ; pharmacology ; Receptors, CXCR4 ; biosynthesis ; genetics ; Stem Cell Factor ; pharmacology ; Thrombopoietin ; pharmacology

Adenoma, Sweat Gland ; metabolism ; Diagnosis, Differential ; Humans ; Keratin-14 ; metabolism ; Keratin-17 ; metabolism ; Keratin-18 ; metabolism ; Keratin-7 ; metabolism ; Keratins ; metabolism ; Papilloma ; metabolism ; Sebaceous Gland Neoplasms ; metabolism ; Skin Neoplasms ; metabolism ; Sweat Gland Neoplasms ; metabolism