Objective To study the therapeutic effects of intermittent hypoxia therapy(IHT)in isolated systolic hypertension(ISH)patients with elevated cerebral blood flow velocity(Vp),and to explore the mechanisms involved.Methods Seventy-six ISH patients with increasing Vp and normal pulsatility index(PI)of the middle ce- rebral artery(MCA)were randomly divided into a therapy group and a control group.IHT was administrated in the therapy group,and air in the control group.The Vp and PI of the MCA and blood pressure(BP)were observed be- fore and after treatment.Results Vp and systolic blood pressure(SBP)were significantly reduced after IHT(P＜0.01)compared with the therapy group's scores betore treatment,but PI and diastolic blood pressure showed no sig- nificant difference.There was no significant change in BP,Vp or PI in the control group before or after treatment. Conclusion IHT has therapeutic effects on ISH by reducing Vp and moderating SBP.

Objective To explore the specificity and efficiency of YFP labeled natural killer (NK) cells through Vav-Cre induced YFP reporter system in mice. Methods ROSA26R-YFP and Vav-Cre mice were crossed, and their YFP and Cre gene double positive progeny were screened by genotyping. The specificity of YFP in hematopoietic cells from im-mune organs including lymph nodes, spleen, thymus and bone marrow were analyzed by flow cytometry. The percentages of YFP positive cells in NK cells from lymph nodes, spleen and bone marrow were also analyzed by flow cytometry. Results A total of 11 double positive mice (ROSA26R-YFP-(+/-)VavCre) were obtained in 17 mouse offspring by crossing ROSA26R-YFP mice with Vav-Cre mice. The percentages of YFP positive cells in immune organs including lymph nodes, spleen, thy-mus and bone marrow were 73.87%± 1.51%, 56.07%± 1.47%, 86.17%± 1.74%and 53.60%± 3.56%, and there were signifi-cant differences compared with the corresponding negative control cells(0.27%±0.01%, 1.33%±0.91%, 0.11%±0.01%and 0.29%± 0.03%, P<0.01). There were no YFP expressions in non-immune organs in double positive mice and in negative control mice (0.72%±0.43%vs 0.92%±0.27%, P>0.05). The positive rates of YFP were significantly higher in NK cells in lymph nodes, spleen and bone marrow (76.94%±0.84%、81.66%±1.18%and 88.92%±0.77%) compared with those of control (P<0.01). Conclusion YFP marked NK cells through Vav-Cre induced YFP reporter system in mice have high specificity and efficiency.

Objective: To study the chemical constituents from the root barks of Lycium chinense. Methods: The chemical constituents of EtOAc fraction from 95% ethanol extract of L. chinense were isolated and purified by chromatography on silica gel, Sephadex LH-20, and ODS. Their chemical structures were identified on the basis of physicochemical properties and spectroscopic data. Results: Twelve compounds were isolated and identified as N-trans-coumaroyltyramine (1), N-trans-feruloyltyramine (2), dihydro-N-caffeoyltyramine (3), apigenin (4), ferulic acid (5), p-hydroxycinnamic acid (6), 3-hydroxy-1-(4-hydroxyphenyl)-1-propanone (7), 3, 4-dihydroxybenzenepropionic acid (8), 3, 4-dihydroxybenzenepropionic acid methyl ester (9), p-hydroxy-benzoic acid (10), 4-methoxy salicylic acid (11), and nicotinic acid (12). Conclusion: Compounds 8 and 9 are two new natural products, and compounds 1, 6, 11, and 12 are obtained from this plant for the first time.

Objective To investigate the effect and possible signal pathway of brain - derived neurotrophic factor (BDNF) on apoptosis of rat embryo brain cells suffering from intrauterine hypoxic - ischemic injury. Methods The uterine arteries of the pregnant rats were clamped for 30 minutes in both experimental group and control group. BDNF(2 ?g) was injected into rats in experimental group while saline was injected into rats in control group through caudal veins. The samples were collected at 24, 48 and 72 hours re-spectively after artery clamping. Neuroapoptosis of different groups induced by ischemic damage was measured by TUNEL assay. The expression of extracellular signal - regulated kinase(ERK)and c - Jun - N - terminal kinase(JNK) were observed by immunohisloche-mistry.Results The number of apoptosis cells after hypoxic - ischemic injury increased progressively with time.The apoptosis cells number in experimental group were much lower in number than those of ischemic control group.The expression of ERK increased while the expression of JNK decreased in experimental group, comparing with that of the ischemic control group, with statistical signif-icance (P

Objective To investigate the relationship between serum adiponectin and metabolic syndrome(MS) in children and adolescents with obesity and nonalcoholic fatty liver disease(NAFLD).Methods Four elementary schools and 4 middle schools were selected from Haidian district in Beijing with representative cluster sampling.Two hundred and eighty obese children(obese group),65 obese children with NAFLD(NAFLD group) and 264 normal weight children(healthy control group) aged 7 to 18 years were recruited from the 8 schools with uncompletely randomized sampling.Data including questionnaire,anthropometric measurements,B type ultrasonographic examination for liver were collected and fasting blood laboratory assay were determined.Variables including triglyceride(TG),adiponectin,alanine aminotransferase(ALT) and aspartate aminotransferase(AST) were skewed distribution and natural logarithmical transformations were performed.Chi-square test for category and multiple binary Logistic regression analysis were used to statistical analysis.Results Body mass index(BMI) and waist circumference(WC) in obese group and NAFLD group were higher than those in healthy control group.All the chi-square tests for trend among the 3 groups were statistically significant(P

[ABSTRACT]OBJECTIVEThe purpose of this study was to analyze the screened miRNAs related to the chemosensitivity for the TPF regimen of hypopharyngeal squamous cell carcinoma by miRNA array, and provide a set of miRNAs that may be useful for the development of novel diagnostic markers and more effective therapeutic strategies from the screened miRNAs.METHODSA total number of 21 patients who underwent TPF induction chemotherapy for primary hypopharyngeal squamous cell carcinoma were recruited for miRNA array analysis. 12 patients are sensitive to chemotherapy, and 9 patients are not. Moreover, the selected putative regulated miRNAs were also validated by RT-PCR in another 24 patients (14 patients are sensitive to chemotherapy, and others are not).RESULTSThere were 24 miRNA significantly differencial to the sensitivity to chemotherapy, and 6 miRNAs were up-regulated in the TPF group while 18 miRNA were down-regulated (P<0.05). To identify typical miRNA, mirfocus 3.0 database selected four miRNAs hsa-miR-211-3p, hsa-miR-4253, hsa-miR-4443, and hsa-miR-193b-3p, which were significant down-regulated in TPF-sensitive group. QRT-PCR further validated that only three miRNA (hsa-miR-4253、hsa-miR-4443、hsa-miR-193b-3p) were under-expressed in TPF-sensitive group of another 24 tissue samples (P<0.05).CONCLUSIONMiRNA hsa-miR-193b-3p, hsa-miR-4253, hsa-miR-4443 were identified in TPF-sensitive tissues by microarrays, and further validated by RT-PCR. These down-regulated miRNAs may act as novel biomarkers to classify TPF sensitivity of hypopharyngeal squamous cell carcinoma patients and will contribute to the understanding of the molecular basis of the chemosensitivity in the disease.

The chemical constituents of 95% ethanol extract of Melastoma dodecandrum were isolated and purified by chromatography on silica gel, Sephadex LH-20, and HPLC, to obtain thirteen compounds eventually. On the basis of their physico-chemical properties and spectroscopic data, these compounds were identified as quercetin (1), quercetin-3-O-β-D-glucopyranoside (2), quercetin-3-O-(6"-O-p-coumaroyl) -β-D-glucopyranoside (3), kaempferol (4), kaempferol-3-O-β-D-glucopyranoside (5), kaempferol-3-O- [2",6"-di-O-(E)-coumaroyl]-β-D-glucopyra-noside (6), luteolin (7), luteolin-7-O-(6"-p-coumaroyl) -β-D-glucopyranoside (8), apigenin (9), apigenin-7-(6"-acetyl-glucopyranoside) (10) , naringenin (11), isovitexin (12), and epicatechin-[8,7-e] -4β-(4-hydroxyphenyl)-3,4-dyhydroxyl-2(3H)-pyranone (13). Eight compounds(3,5,6,8-11 and 13) were obtained from M. dodecandrum for the first time.
Apigenin ; analysis ; Chromatography ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Flavanones ; analysis ; Flavonoids ; analysis ; chemistry ; Glycosides ; analysis ; chemistry ; Kaempferols ; analysis ; Luteolin ; analysis ; Magnoliopsida ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analysis ; Silica Gel

BACKGROUND: Osteoporosis is a genetic disease associated with many enes. To date, the genes that regulate bone mass are incompletely defined.OBJECTIVE: To investigate the relationship between polymorphisms of steoprotegerin (OPG) gene promoter with bone mineral density (BMD) in remenopausal and postmenopausal women.DESIGN: Prospective study.SETTING: Peking Union Medical College Hospital.PARTICIPANTS: In July 2002, 495 Han nationality women selected from Peking Union Medical College Hospital were non-related volunteers and gave their informed consent prior to the study, which included 306 premenopausal women aged 20-39 years, 189 postmenopausal women aged 50-84 years.METHODS: ① BMD measurement: BMD was measured at the Lumbar Spine and Femoral Neck, trochanter, Ward's triangle by dual-energy X-ray absorptiometry. ② Genotyping: Whole blood genome DNA was extracted by QIAGEN DNA extraction kit. The PCR product and the result of endonuclease digest were confirmed by sequencing (Bioasia Biotechnology,Shanghai, China). The impact of the polymorphisms on BMD was also investigated using multiple Logistic regression.MAIN OUTCOME MEASURES: ① Distribution of OPG genotypes and the relationship with BMD. ② Association between OPG polymorphisms and osteoporosis.RESULTS: All 495 subjects were involved in the final analysis. ① These polymorphisms were in Hardy-Weinberg equilibrium (χ2= 0.056 -0.222, P＞ 0.05). The frequencies of genotypes of these subjects were as follows: AA (70.1%), AG (26.9 %), GG (3.0 %) for 163A→G polymorphism; TT (71.3 %), TG (25.9 %), GG (2.8 %) for 245T→G polymorphism. BMD was lower in premenopausal women with GG +AG genotype than AA genotype for 163A→G polymorphism, so did GG+TG genotype than TT genotype for 245T→G polymorphism. But there was no significant difference. BMD was lower in postmenopausal women with AG+GG genotype than AA genotype for 163A→G polymorphism at Lumbar Spine 2-4, Femoral Neck, Ward's triangle and Trochanter (P ＜ 0.05). For 245T→G polymorphism, BMD of postmenopausal women with TG+GG genotype was lower at Femoral Neck,Ward's triangle and Trochanter than TT genotype (P ＜ 0.05). For 245T→G polymorphism, BMD of postmenopausal women with TG+GG genotype was lower at Femoral Neck, Ward's triangle, and Trochanter than TT genotype (P ＜ 0.05). ② Age, weight, height, years since menopause, and 163A→G/245T→G genotypes were sewed as covariates. AG+GG genotype was contributed to low BMD at Lumbar Spine 2-4 and Ward's triangle (OR =2.045, OR=2.956, P ＜ 0.05, 95% CI 1.05-6.7). TG+GG genotype was risk factor for osteoporosis at Lumbar Spine 2-4, Ward's triangle,and Trochanter (OR=2.059, OR=2.859, OR=2.123, P ＜ 0.05, 95% CI 1.04-6.5).CONCLUSION: BMD was lower in postmenopausal women with the variant G allele for 163A→G and 245T→G polymorphisms at Femoral Neck,Ward's triangle, and Trochanter. The variant allele G may associate with lower BMD in postmenopausal women.

By using a yeast two-hybrid system, a yeast two-hybrid bait vector was constructed and identified for screening of the HPV18 E6-interacting proteins, and its effects on the growth of yeast cells and the activation of reporter genes were investigated. Total mRNA extracted from Hela cells was reversely transcribed into cDNA. Fragment of HPV18 E6 cDNA was amplified using RT-PCR and directly ligated to the pGBKT7 vector. The recombinant plasmid was confirmed by restriction endonuclease analysis and DNA sequencing. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector were transformed into the yeast cell AH109, respectively. After they were cultured respectively in YPDA liquid medium and nutrition-deficient culture medium, their toxicity and transcriptional activation were tested by both the phenotype assay and the color assay. The bait plasmid HPV18 E6 was successfully obtained. After being cultured in YPDA liquid medium for 16h, the A (600 nm) values of two yeast fluids were 0.98+/-0.03 and 0.99+/-0.02, respectively. The recombinant pGBKT7-HPV18 E6 plasmid and empty pGBKT7 vector could grow to white colonies on SD/-Trp/X-alpha-gal plates, while no colony could survive on SD/-His/-Trp/X-alpha-gal, SD/-Ade/-Trp/X-alpha-gal plates, indicating that the bait plasmid pGBKT7-HPV18 E6 was constructed successfully and expressed correctly, and could not activate the transcription of reporter gene alone. The yeast two-hybrid GAL4 system 3 can be utilized to find HPV18 E6 interacting proteins.

Objective To explore the effects of Chinese herbal compound, Tengmei decoction, on type II collagen-induced arthritis ( CIA) in rats, and to examine the changes of arthritis index ( AI) , limb swelling, joint tissue inflammatory infiltration, and the effects on immune-inflammatory factors.Methods Sprague-Dawley rat models of arthritis were successfully established by intradermal injection of type II collagen and Freund’ s complete adjuvant.The model rats were randomly divided into model group, positive drug group, and high-and low-dose Chinese medicine groups, 6 rats in each group.The intervention and treatment period was 12 weeks.To measure weekly the anteroposterior and transverse diameters of the rear ankles and wrists, the transverse diameter of the claw foot palm pad, the thickness and the highest point width of hind limb plantar joint swelling, and to evaluate the integrated scores of joints and limbs swelling using a vernier caliper.Results ①Compared with the normal group, the total arthritis scores and hind limbs AI scores of the model group were significantly increased ( P <0.05 or P <0.01 ) .The left forelimb AI scores were significantly increased during 10 -12 weeks ( P <0.05 ) .The anteroposterior and transverse diameters of the left hind limb, the thickness of the highest point measurement of the left hind foot pad metatarsal were significantly increased ( P<0.05 or P<0.01) in different time periods between 1-12 weeks.Compared with the model group, the total scores and the left hind limb joints AI scores of the high-and low-dose drug groups were decreased after 6 weeks (P<0.05).②Compared with the normal control group, levels of mRNA transcription and protein expression of IL-6 and TNF-αwere significantly up-regulated ( P<0.01 ) in the model group.Compared with the model group, the levels of mRNA transcription and the expression of IL-6 and TNF-αproteins were significantly down-regulated in the positive group and Chinese medicine groups ( P <0.01 ) .③ Histological examination showed that the low-dose TCM significantly improved the CIA synovial hyperplasia and inflammatory cell infiltration.Conclusions The molecular mechanism of Chinese herbal compound Tengmei decotion in improving joint pathological injury of CIA rat models may be related to its inhibitory effect on the high expression of immune-inflammatory factors in the synovial tissue of CIA rats.