In the current study, nine flavonoids were isolated and purified from 75% ethanol extract of Selaginella uncinata (Desv.) Spring by column chromatographic techniques over macroporous resin, polyamide, silica gel, Sephadex LH-20 and pre-HPLC. On the basis of their physico-chemical properties and spectroscopic data analyses, these compounds were elucidated as cirsimarin (1), nepitrin (2), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), apigenin-6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (4), apigenin-7-O-β-D-glucopyranoside (5), 2,3-dihydroamentoflavone (6), 4'-O-methylamentoflavone (7), 2,3-dihydro-4'-O-methyl-amentoflavone (8), and 2,3,2",3"-tetrahydron-4'-O-methyl-robustaflavone (9). Compounds 1-5 belong to flavonoid glycosides and were isolated from the genus Selaginella for the first time.
Flavonoids ; analysis ; Selaginellaceae ; chemistry

OBJECTIVETo establish a HPLC-based method for simultaneous determination of 2 classes of compounds (flavonoids and chromones) and 6 their effective components,(including prin-O-glucosylcimifugin, cimifugin, 4'-O-beta-D-glucosyl- 5-O-methylvisamminol, quercetin, sec-o-glucosylhamaudol and formononetin), in Yupingfeng Decoction.

METHODSHPLC-based separation of the agents was performed on Agilent Extend-C(18) column (4.6 mm x 250 mm, 5 microm) at 25 degrees with the mobile phase of MeOH-1% acetic acid water solution (gradient elution), flow rate of 0.8 ml/min and detection wavelength of 254 nm.

RESULTS AND CONCLUSIONHPLC allowed simultaneous quantitative determination of the 6 components in Yupingfeng Decoction, and they showed good linear relationships when their sample amount ranged 90-1810 ng, 97-1940 ng, 190-1906 ng, 105-3144 ng, 88-2625 ng and 109-3279 ng, respectively, with correlation coefficients all beyond 0.9999 and average recovery rates of 98.2%, 99.1%, 97.3%, 97.8%, 98.8% and 99.2%, respectively. This simple and convenient method accommodated a broad linear range with high sensitivity and precise and reproducible results.

Chromatography, High Pressure Liquid ; methods ; Chromones ; analysis ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; Isoflavones ; analysis ; Quercetin ; analysis ; Reproducibility of Results

The opening of laboratories in universities is one of important parts in teaching reform and it is necessary for bringing up high-qualified students.Combined with the practical teaching,we have a primary discussion with the problems of the laborato- ry opening and put forward some suggestions and measures.

The cytokines of acute leukemia (AL) patients have certain expression patterns, forming a complex network involved in diagnosis, progression, and prognosis. We collected the serum of different AL patients before and after complete remission (CR) for detection of cytokines by using an antibody chip. The expression patterns of cytokines were determined by using bioinformatics computational analysis. The results showed that there were significant differences in the cytokine expression patterns between AL patients and normal controls, as well as between acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). In confirmatory test, ELISA revealed the expression of uPAR in AL. Moreover, the bioinformatic analysis showed that the differentially expressed cytokines among the AL groups were involved in different biological behaviors and were closely related with the development of the disease. It was concluded that the cytokine expression pattern of AL patients is significantly different from that of healthy volunteers. Also, differences of cytokine expression patterns exist between AML and ALL, and between before and after CR in the same subtype of AL, which holds important clinical significance for revealing disease progression.

OBJECTIVETo observe the effect of paiqian chewing tablet (PQCT) on lead discharging and health in children.

METHODSAdopting self-control and inter-group control method, 94 children with blood lead level exceeding 100 microg/L were randomly divided into the observed group and the control group. The observation period for both groups was 30 days.

RESULTSAt the 20th and 30th day of treatment, the urinary lead output in the observed group was significantly higher than that in the control group (P < 0.05, P < 0.01), and showed significant difference as compared with that before treatment (P < 0.05). Besides, the total amount of urinary lead discharging in the observed group was significantly more than that in the control group (P < 0.05).

CONCLUSIONPQCT has markedly lead discharging improvement action with no influence on urinary calcium and zinc excretion. As all the routine indexes of blood and urine ranged within the normal extent, it demonstrated that PQCT was harmless to the health of observed individual.

Child ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Lead ; blood ; urine ; Lead Poisoning ; drug therapy ; Male ; Phytotherapy ; Tablets

OBJECTIVETo investigate the infection rate and subtypes of human papilloma virus(HPV) in patients with oropharyngeal squamous cell carcinoma (OSCC) and analyze the clinicopathologic features of patients with or without HPV infection.

METHODSA total of 66 biopsy or surgical specimens of OSCC archived in the Pathology Department of Chinese Academy of Medical Sciences were analyzed by polymerase chain reaction (PCR), and the generic amplification products were detected by DNA enzyme immunoassay (DEIA) and typed by reverse hybridization line probe assay.

RESULTSHPV-DNA was detected in 11 (16.7%) of all specimens. Among them, 7 were infected with HPV-16,and the remaining 4 patients were infected with HPV-16/11, HPV-35, HPV-58/52, and HPV-33/52/54, respectively. HPV-16 was detected in 72.7% of all positive specimens. There were more females in HPV-positive group than HPV-negative group (36.4% vs. 1.8%,P=0.002). Patients with HPV-positive tumors were more likely to be non-smokers (36.4% vs. 0,p=0.001) and non-drinkers (45.5% vs. 1.8%,p=0.001) than those with HPV-negative tumors. The proportion of moderately or poorly differentiated tumors was higher in HPV-positive patients than HPV-negative patients (81.8% vs. 63.7%), although without statistical significance (p=0.409). No difference was observed in T classification, N classification, and overall tumor stage.

CONCLUSIONSHPV infection rate was 16.7% in this cohort. HPV-positive OSCC has its unique etiologic and clinicopathological characteristics.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; virology ; DNA, Viral ; isolation & purification ; Female ; Humans ; Male ; Middle Aged ; Oropharyngeal Neoplasms ; virology ; Papillomaviridae ; classification ; isolation & purification ; Papillomavirus Infections ; virology

The triterpenoids of Dichrocephala benthamii were investigated by means of silica gel, Sephadex LH-20 and semi-preparative HPLC. Nine triterpenoids were isolated from D. benthamii. By analysis of the EI-MS, NMR spectra and comparison to the data reported in literatures, the structures of these compounds were determined as β-amyrin formiate (1), β-amyrin acetate (2), β-amyrenol (3), β-amyrone (4), 3β-hydroxy-olean-11, 13 (18)-diene (5) , Δ12-oleanene (6) , friedelin (7), dammaradienyl acetate (8), epi-friedeband (9), respectively. Compounds 1-8 were isolated for the first time form this genus, compound 9 was isolated for the first time from this plant, whereas β-amyrin formiate (1) was a new natural product.
Asteraceae ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo transfect pcDNA3.1/hTM plasmids containing human thrombomodulin (hTM) gene into human umbilical vein endothelial cells (HUVECs), and investigate the expression of hTM and anticoagulating function of transfected HUVECs.

METHODSHUVECs were transfected with pcDNA3.1/hTM by lipofectin. Expression of hTM mRNA was determined by semi-quantitative RT-PCR, hTM antigen on HUVECs membrane by immunohistochemistry, and activated protein C (PC) in HUVECs by chronometry. By using a semiautomatic coagulator, the effect of the reacting liquid from transfected HUVECs mixed with PC from normal peripheral blood was assayed.

RESULTSAbout 10% HUVECs were transfected by pcDNA3.1/hTM with high-level hTM mRNA and protein expression. Activated PC produced by pcDNA3.1/hTM group, pcDNA3.1(+)/neo group and untransfected group was (2.80 +/- 0.43) microg/ml, (0.75 +/- 0.08) microg/ml and (0.85 +/- 0.11) microg/ml, respectively. APTT was (51.68 +/- 2.73) s, (38.38 +/- 2.44) s, (39.65 +/- 2.39) s, (33.93 +/- 1.73) s and (34.60 +/- 1.86) s and PT was (21.89 +/- 1.66) s, (20.56 +/- 1.74) s, (20.42 +/- 2.04) s, (19.57 +/- 1.36) s and (20.16 +/- 1.35) s in pcDNA3.1/hTM group, pcDNA3.1(+)/neo group, untransfected group, inactivating PC group and control, respectively.

CONCLUSIONSThe pcDNA3.1/hTM plasmid could be transfected into endothelial cells and expressed biologically functioning hTM protein on HUVECs membrane. Activated PC could inhibit intrinsic coagulation pathway obviously with slight effect on extrinsic pathway.

Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; cytology ; Gene Expression Regulation ; Genes, Transgenic, Suicide ; Humans ; In Vitro Techniques ; Partial Thromboplastin Time ; Plasmids ; Protein C ; Prothrombin Time ; Thrombomodulin ; genetics ; Transfection ; Umbilical Veins ; cytology

Objective To evaluate the advantage of homemade negative pressure device combined with nano-silver dressing for promoting the healing of infected incision in rats,and explore its clinical curative effect.Methods In-fected incision model rats were randomly divided into conventional treatment group,and simple pressure suction group,pressure suction combined with silver ion dressing group.The healing time and healing area of rats in each group after treatment were evaluated,immunohistochemical and fluorescent quantitative analysis of inflammatory factors in incisional wound tissue were performed.Three methods were applied to patients with surgical site infec-tion(SSI),granulation coverage time,granulation recovery time,and incision healing time of three groups of pa-tients were compared.Results Immunohistochemistry and its IOD value,the relative mRNA expression levels of TNF-α,IL-2,and IL-8 in rat wound tissue treated with pressure suction combined with silver ion dressing were all inferior to conventional treatment group and simple negative pressure suction group,difference was statistically sig-nificant (P < 0.05);in clinical application,wound healing time,postoperative C-reactive protein level,and pain as-sessment scores in patients treated with pressure suction combined with silver ion dressing were all superior to con-ventional treatment group and simple negative pressure suction group,difference were all statistically significant (all P < 0.05).Conclusion Compared with conventional treatment method,pressure suction with silver ions dressing treatment can more effectively control SSI,reduce local inflammation of incision,and promote incision healing.

Objective To explore the relationship between HBV infection and the genotypes and allele frequencies of CⅡTA G-944C gene polymorphism in three minority populations(Jinuo,Dai and Aini population)in Xishuangbanna district,Yunnan province.Methods Polymerase chain reaction and sequencing method were used to study the genotypes and allele frequencies distributions of CⅡTA G-944C gene polymorphism in those three populations.Relationship between the genotypes distribution and HBV infection results were also analyzed.Results The rates on HBV infection and HBsAg carrier status in Aini minority population were 89.2% and 16.3%,which were significantly higher than in Jinuo(27.9% and 3.9%,χ~2=135.196 and 10.361,P=0.000 and 0.001)and Dai population(44.9% and 6.6% χ~2=96.783 and 8.748,P=0.000 and 0.003)while among Aini population it was significantly different with the other two minority populations.The CC genotype and C allele frequencies were more distributed in Aini population than in the other two minority populations.In contrast,the GG genotype and G allele frequencies were lower than the other two minority populations,with χ~2 rates between Aini and Jinuo population were 11.841 and 12.208 and the P as 0.003 and 0.000 respectively while the χ~2 rates between Ami and Dai population were 23.902 and 20.220 with P value as 0.000 and 0.000.The genotypes frequencies of CⅡTA G-944C was significantly different in the infected individuals(IF)group and health control(HC)group in Jinuo population(χ~2=6.150 and 4.911,P=0.046 and 0.027).When compared with HBsAg+ group and HBsAg~- group,the genotypes and allele frequencies were different in Aini population and the total three minority populations(χ~2 rates in Jinuo minority were 8.650 and 5.034 with P values as 0.013 and 0.025).However,the χ~2 rates in the whole population were 13.047 and 9.416 with P values as 0.001 and 0.002,respectively.The distribution of CC genotype and C allele gene in HBsAg~+ group was increasing.Data from non-condition logistic regression analysis and adjusting for confounding factors,the HBsAg~+ group had a significantly increase of HBsAg~- group under the C allele Recessive Model(P=0.000;OR=2.964;95% CI:1.609-5.460).Conclusion The genotypes and allele frequencies distribution of CⅡTA G-944C were different in the three ethnic populations.Polymorphism of this gene was closely associated with HBsAg carrier.The CC genotype patients were more easily to become HBsAg carrier.