Chemical constituents of Helleborus thibetanus / 中国中药杂志

The chemical constituents of Helleborus thibetanus were isolated and purified by silica gel column chromatography, Sephadex LH-20 gel column chromatography, and semi-preparative RP-HPLC, and the structures of all compounds were identified by modern spectrographic technology(MS, NMR). The MTT method was used to measure the cytotoxicity of compounds 1-8. Twelve compounds were isolated from the roots and rhizomes of H. thibetanus and were identified as(25R)-22β,25-expoxy-26-[(O-β-D-glucopyranosyl)oxy]-1β,3β-dihydroxyfurosta-5-en(1), β-sitosterol myristate(2), β-sitosterol lactate(3), β-sitosterol 3-O-β-D-glucopyrannoside(4), 4,6,8-trihydroxy-3,4-dihydronaphthalen-1(2H)-one(5), 1,3,5-trimethoxybenzene(6), 7,8-dimethylbenzo pteridine-2,4(1H,3H)-dione(7), 1H-indole-3-carboxylic acid(8), p-hydroxy cinnamic acid(9), lauric acid(10), n-butyl α-L-arabinofuranoside(11) and methyl-α-D-fructofuranoside(12), respectively. Among them, compound 1 is a new compound and named thibetanoside L; compounds 2, 5-8, 11 are first isolated from the family Ranunculaceae; compound 12 is isolated from the genus Helleborus for the first time. The results of MTT assay showed that the IC_(50) values of compounds 1-8 against HepG2 and HCT116 cells were greater than 100 μmol·L~(-1).

Five new polyhydroxylated furostanol saponins from the rhizomes of Tupistra chinensis / 中国天然药物

Five new polyhydroxylated furostanol saponins were isolated from the roots and rhizomes of Tupistra chinensis, and their structures were determined as tupistrosides J-N (1-5), together with four known furostanol saponins (6-9), on the basis of physico-chemical properties and spectral analysis. Among them, compounds 3 and 5 showed cytotoxicity against human cancer cell lines SW620 with IC values of 72.5 ± 2.4 and 77.3 ± 2.5 μmol·L, respectively. Compound 4 showed cytotoxicity against human cancer cell line HepG2 with IC value of 88.6 ± 2.1 μmol·L.

Steroidal constituents from Helleborus thibetanus and their cytotoxicities / 中国天然药物

Thibetanosides E-H (1-4), four new steroidal constituents including three rare sulfonates (2-4), were isolated from the roots and rhizomes of Helleborus thibetanus, together with nine known steroidal compounds (5-13). Their structures were elucidated by detailed spectroscopic analysis, including 1D and 2D NMR techniques and chemical evidence. In this study, compounds 2-13 were evaluated for their cytotoxic activities against HCT116, A549 and HepG2 tumor cell lines in vitro. Among them, compound 8 (thibetanoside C) showed cytotoxicities against A549 cells(IC 39.6 ± 1.9 μmol·L) and HepG2 cells(IC 41.5 ± 1.1 μmol·L), respectively. Compound 9 (23S, 24S)-24-[(O-β-D-fucopyranosyl)oxy]-3β, 23-dihydroxy-spirosta-5, 25(27)-diene-1β-ylO-(4-O-acetyl- α-L-rhamnopyranosyl)-(1→2)-O-[β-D-xylopyranosyl-(1→3)]-α-L-arabinopyranoside) showed cytotoxicity against HCT116 cells(IC 33.6 ± 2.1 μmol·L).

Physicochemical properties of hydroxyapatite/dicalcium phosphate dehydrate bone cement / 中国组织工程研究

BACKGROUND:β-tricalcium phosphate (β-TCP) and monocalciumphosphate monohydrate (MCPM) are traditionally considered as reactants for dicalcium phosphate dehydrate (DCPD) bone cement,but little is reported on the hydroxyapatite (HA) as a reactant.OBJECTIVE:To verify whether HA and MCPM can be used to prepare DCPD bone cement and to explore the physicochemical properties.METHODS:The HA and β-TCP were prepared by wet chemical precipitation method,and mixed with appropriate proportion of MCPM.Then,the HA-DCPD and β-TCP-DCPD were obtained by adding a proper amount of curing water.The composition and structure of the two materials were analyzed by X-ray diffraction,the morphology was observed by scanning electron microscope,and the mechanical strength was tested by Instron5567 universal material test machine.These two kinds of materials were placed in simulated body fluid for detecting the weight loss ratio,soaked for 14 days and taken out for X-ray diffraction and scanning electron microscope detection.RESULTS AND CONCLUSION:X-ray diffraction findings indicated that these two kinds of materials both belonged to high-purity DCPD bone cement.Under the scanning electron microscope,β-TCP-DCPD bone cement had dense crystal structure,with less pore number;however,the HA-DCPD bone cement presented with finer grains,loose structure,and higher pore number.With the increase of curing time,the mechanical strength of two kinds of bone cements was correspondingly increased,but the compressive strength of β-TCP-DCPD bone cement was significantly higher than that of HA-DCPD bone cement (P ＜ 0.05).In the simulated body fluid,the weight loss ratio of β-TCP-DCPD bone cement was significantly lower than that of HA-DCPD bone cement (P ＜ 0.05).At 14 days after soaking in the simulated body fluid,a layer of spherical particles that was formed on the surface of both materials was identified as hydroxyapatite by scanning electron microscope observation and X-ray diffraction analysis.In summary,HA-DCPD bone cement has good biodegradability,excellent bioactivity and bone conductivity,but poor mechanical properties.

Synthesis of new 4-anilinoquinazoline analogues and evaluation of their EGFR inhibitor activity / 药学学报

Thirteen of 4-anilinoquinazoline derivatives with imine groups at position 6 of quinazoline ring were synthesized and their antitumor activities were evaluated by MTT assay and Western blotting analysis. Among these compounds, 13a-131 were reported first time. The MTT assay was carried out on three human cancer cell lines (A549, HepG2 and SMMC7721) with EGFR highly expressed. Among the tested compounds, 13i and 13j exhibited notable inhibition potency and their IC50 values on three cell lines were equivalent to or less than those of gefitinib. Compound 14, without imine group substituted, displayed excellent inhibitor potency only on A549 cell line. Compounds 14 and 13j were chosen to perform Western blotting analysis on A549. The results showed that both of the compounds could inhibit the expression level of phosphorylated EGFR remarkably. It was concluded that the inhibitor potency of compound 14 was almost equivalent to that of gefitinib and the inhibitor potency of 13j was better than that of gefitinib.

Protection of salvianolate against atherosclerosis via regulating the inflammation in rats / 华中科技大学学报（医学）（英德文版）

Inflammation plays an essential role in the pathophysiology of atherosclerosis. Our study was aimed to investigate whether salvianolate, a novel water-soluble phenolic compound of Danshen, alleviates atherosclerosis via regulating the inflammation in rats. High fat diet feeding plus vitamin D3 injection was used to induce atherosclerosis in rats. Salvianolate (60, 120 or 240 mg/kg) or placebo was given to atherosclerotic rats. The plasma lipids, interleukin 6 (IL-6) and C reactive protein (CRP) were measured by ELISA. CD4+CD25+Foxp3+ cells were determined by flow cytometry. Histological changes were examined by hematoxylin and eosin staining. The results showed that the levels of plasma IL-6 and CRP were elevated in the rats fed on high fat diet, and the histological analysis demonstrated the successful establishment of atherosclerosis models. Treatment with salvianolate alleviated the atherosclerotic process and decreased the levels of plasma IL-6 and CRP. Also the number of CD4+CD25+Foxp3+ cells was increased in salvianolate-treated rats. It was concluded that salvianolate could treat atherosclerosis via modulating the inflammation at cytokine and cell levels.

Protection of salvianolate against atherosclerosis via regulating the inflammation in rats / 华中科技大学学报（医学）（英德文版）

Inflammation plays an essential role in the pathophysiology of atherosclerosis. Our study was aimed to investigate whether salvianolate, a novel water-soluble phenolic compound of Danshen, alleviates atherosclerosis via regulating the inflammation in rats. High fat diet feeding plus vitamin D3 injection was used to induce atherosclerosis in rats. Salvianolate (60, 120 or 240 mg/kg) or placebo was given to atherosclerotic rats. The plasma lipids, interleukin 6 (IL-6) and C reactive protein (CRP) were measured by ELISA. CD4+CD25+Foxp3+ cells were determined by flow cytometry. Histological changes were examined by hematoxylin and eosin staining. The results showed that the levels of plasma IL-6 and CRP were elevated in the rats fed on high fat diet, and the histological analysis demonstrated the successful establishment of atherosclerosis models. Treatment with salvianolate alleviated the atherosclerotic process and decreased the levels of plasma IL-6 and CRP. Also the number of CD4+CD25+Foxp3+ cells was increased in salvianolate-treated rats. It was concluded that salvianolate could treat atherosclerosis via modulating the inflammation at cytokine and cell levels.

Establish and optimization of real-time fluorescent reverse transcription loop-mediated isothermal amplification for detection of avian influenza H5 hemagglutinin gene / 病毒学报

H5 subtype avian influenza (AIV-H5) is a major causative agent of animalloimia a rapid and sensitive molecular biological diagnosis is crucial to the control program of AIV-H5. AIV-H5 real-time fluorescent reverse transcription loop-mediated isothermal amplification (qRT-LAMP) was established by means of heat treatment of the samples. The sensitivity, specificity and repeatability of this method were assessed and the performance of Calcein,SYBR Green I,HNB,SYTO 81 in colorimetric detection was comparatively analyzed to screen the optimum dye. The results showed the sensitivity of this method was 100 times higher than that of standard real-time fluorescent RT-PCR, and the detection limit was one copy of the gene per reaction. This method had no cross-reactivity with other common avian respiratory tract infectious disease-related pathogens such as IBV and NDV. The present study suggested Calcein was the optimum dye. Small-scale tests suggested this method was reliable for survey monitoring of AIV-H5 on the spot, indicating its potential applications in field investigation.

Synthesis and anti-tumor activity of paeonol and its derivatives / 药学学报

A series of paeonol derivatives have been synthesized by simple acylation and etherification of the paeonol. Anti-tumor activities of the synthesized compounds were evaluated against HeLa and MCF-7 cells lines in vitro by the standard MTT assay. It was found that the derivatives were more active against HeLa than MCF-7. The results also indicated that 4-methoxy group is the synergistic group of paeonol's anti-tumor activity and ketone carbonyl side chain is essential functional group of paeonol's anti-tumor activity. Compound 2d had stronger antiproliferative activities than paeonol against HeLa and MCF-7 cell lines with IC50 values of 2.67 and 4.74 micromol x L(-1) respectively. The results showed that paeonol derivatives were worth to be intensively studied further.

Comparison of the quantities and subset distributions of natural killer cells among different races / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Natural killer (NK) cells play critical roles in host immune defense, while the quantities and subset distributions may vary among different races. To address the difference, we compared these variables among Chinese Han, the Caucasians and the Blacks. The study may provide critical background information for both basic research and clinical investigation.</p><p><b>METHODS</b>Blood samples collected from populations of different races were tested within 12 hours after collection and subsets of NK cells were characterized using flow cytometry.</p><p><b>RESULTS</b>The absolute NK count in the Chinese Han was significantly higher than that in the Caucasian. The Han and Caucasian groups showed higher percentages of cytotoxic subset compared to that of the Black group. The percentage of cytokine-producing subset of Chinese Han group was lower than that of Caucasian and Black groups. Black group had a higher percentage of function-unknown NK subset than that of the Han and Caucasian groups.</p><p><b>CONCLUSION</b>Our data indicated that NK cell count and the distribution of different subsets varied among different races, which should be taken into consideration in related investigations.</p>

Isolation of a Bacterium with High Product of Alkaline Mn-superoxide Dismutase,Geneclone and Sequence Analysis / 中国生物工程杂志

A high-product superoxide dismutase(SOD)bacterial strain was first obtained from the soil around the warm spring of Changbai Mountain,which is alkaline-resistant.The nature analysis by the inhibition of H2O2 and chloroform-ethanol indicated that the SOD was Mn-SOD.It belongs to bacillus genus by the modal character,physiological and biochemical character.The phylogenetic analyses based on 16SrDNA sequence indicated that the strain 110-2 was closed to Bacillus sp.MO6 with 100% identity and 99% with Bacillus licheniformis.According to the sodA sequence of Bacillus licheniformis and conservative regions of many kinds of bacillus,which were published in GenBank,two pairs of primers were designed and the complete sequence of Mn-SOD(600bp)and the core fragment(430bp)were amplified by PCR techniques.The recombinant plasmid pMD18-SOD was built successfully.

Statistical analysis of clinicopathologic characteristics of 1075 cases with colonic cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathologic characteristics of colonic cancer.</p><p><b>METHODS</b>All clinical data of surgical cases from hospital registry from 1982 to 2002 were analyzed using SPSS 11.0 software.</p><p><b>RESULTS</b>There were 1075 patients with colonic cancer including 573 males and 502 females. The median age of the patients was 58 years. 90.8% of the patients aged over 40 years . The proportion of right colonic cancer was 64.7%. Well-differentiated and moderately differentiated adenocarcinoma accounted for 69.8%, and mucous carcinoma 13.6%. The proportions of early stage I, advanced stage II, III and IV cancer were 8.1%, 45.0%, 26.5% and 20.3% respectively. The 5-, 10-year survival rates were 61.9% and 53.0% respectively.</p><p><b>CONCLUSIONS</b>The risk for colonic cancer is significantly increased in patients over 40 years. The frequency of right colonic cancer is higher than that of left colonic cancer. The overall survival rate is high.</p>

Effects of raloxifene hydrochloride on bone mineral density, bone metabolism and serum lipids in Chinese postmenopausal women with osteoporosis: a multi-center, randomized, placebo-controlled clinical trial / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Raloxifene has been approved for prevention and treatment of postmenopausal osteoporosis in Caucasian women. It also has some positive effects on serum lipids in Caucasians. The objective of this study was to determine the effect of raloxifene hydrochloride on lumbar spine and total hip bone mineral density (BMD), bone metabolism, and serum lipids in Chinese postmenopausal women with osteoporosis.</p><p><b>METHODS</b>This was a multi-center, randomized, double-blind, placebo-controlled clinical trial in which 204 postmenopausal Chinese women with osteoporosis were assigned to receive raloxifene (60 mg) or placebo treatment daily for 12 months. BMD, serum bone metabolism markers, and serum lipids were measured before and after drug administration. BMD was measured by Dual-Energy X-Ray Absorptiometry (DEXA) and bone metabolism markers were analyzed by one-step enzyme-linked immunosorbent assay. Serum lipids were measured by enzymatic analysis.</p><p><b>RESULTS</b>At the end of the 12-month study, lumbar spine BMD increased in both groups with a mean increase of (3.3 +/- 4.8)% in the raloxifene group and (1.0 +/- 4.9)% in the placebo group (P < 0.001). There was a mean increase in total hip BMD of (1.4 +/- 4.8)% in the raloxifene group and a mean decrease of (0.9 +/- 5.0)% in the placebo group (P < 0.001). No subject in the raloxifene group had a new vertebral fracture and 5 placebo subjects had new fractures (P > 0.05). In the raloxifene group, the median decreases in the biochemical markers of bone metabolism serum osteocalcin and C-telopeptide were 41.7% and 61.5%, respectively. These changes were statistically significant compared with those in the placebo group (10.6% and 35.6%, P < 0.001, respectively). Both total cholesterol and low-density lipoprotein cholesterol decreased significantly in the raloxifene group compared with those in the placebo group (P < 0.001, respectively) and there was no significant effect of raloxifene on high-density lipoprotein cholesterol and triglycerides compared with placebo.</p><p><b>CONCLUSIONS</b>Raloxifene 60 mg/d for 12 months significantly increases lumbar spine and total hip BMD, significantly decreases bone turnover, and has favourable effects on serum lipids in Chinese postmenopausal women with osteoporosis.</p>

Some problems of postmenopausal osteoporosis prevention with hormone replacement therapy / 中国医学科学院学报

Estrogen deficiency is the one of pathogenetic causes of postmenopausal osteoporosis. Exogenous estrogen may prevent postmenopausal bone loss, so to prevent postmenopausal osteoporosis and osteoporotic fracture. The effect of estrogen on postmenopausal syndrome can not be substituted with other medicine. However, the attention should be paid to the safety of hormone replacement therapy (HRT) during its long term use, and HRT should be used individually.