The chemical consitituents from cytotoxic fraction of the Callicarpa nudiflora extract were isolated and purified by a combination of HP-20 macroporous resin, silica gel and Sephadex LH-20 column chromatographies. The structures were elucidated on the basis of the spectroscopic data and comparison of their spectroscopic data with reported data. The cytotoxicity was evaluated by the MTT assay. The 50% and 70% EtOH elutions of EtOH-extract showed significant cytotoxic activities, leading to the isolation of twelve compounds, which were identified as luteoloside(1), lutedin-4'-O-β-D-glucoside(2), 6-hydroxyluteolin-7-O-β-glucoside(3), lutedin-7-O-neohesperidoside(4), rhoifolin (5), luteolin-7, 4'-di-O-glucoside (6), forsythoside B (7), acteoside (8), alyssonoside (9), catalpol(10), nudifloside(11), and leonuride(12). Compounds 3-6, 10 and 12 were isolated from this genus for the first time, and compound 9 was isolated from this plant for the first time. The cytotoxicity assay demonstrated that flavonoids 1-6, in various concentrations, showed monolithic proliferation inhibitory activities against Hela, A549 and MCF-7 cell lines. Compounds 3, 5 and iridoid glycoside 11 possessed higher cytotoxicacivities. In short, flavonoids are the main components of cytotoxic extract from C. nudiflora, while phenylethanoid glycosides are the predominant ingredient but inactive to cancer cell lines. In addition, the minor iridoid glycoside expressed weak cytotoxic activity.
Callicarpa ; chemistry ; Cell Proliferation ; drug effects ; Cytotoxins ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; MCF-7 Cells ; Molecular Structure

Objective Imatinib mesylate (IM) is the most active agent in treating chronic myeloid leukemia (CML). 5-Aza-2-deoxycytidine (DAC) is a cytosine analogue that inhibits DNA methylation and the activity in myeloid leukemia. Therefore,we investigated combining these two drugs in human leukemia cell line K562. Methods The effects of IM and DAC was examined in K562 cells including cell viability using MTT method,cell cycle phase and cell death using flow cytometric (FCM),and the expression of bcr-abl mRNA by RT-PCR. Results Both DAC and IM resulted in time and concentration-dependent induction of cell death. DAC and IM in combination produced a greater inhibition of growth against K562 cells (F =43.947,165.580,321.193,296.101,P<0.05). The main effect and interaction between two drugs was statistically significant (F = 202.759,168.457,417.538,P <0.001) after 24 h,48 h,72 h and a greater reduction in expression of bcr-abl mRNA than either agent alone. The difference was statistically significant (F =71.981,P <0.05). The number of G1 phase cells were increased significantly when induced by single agent. 48 h incubation with IM 0.2 μmol/L alone or combined with DAC 4 μmol/L showed 6.7 %,8.4 % pre-apoptosis cells,respectively. After incubation for 48 h with DAC 4 μmol/L,the expression of mRNA were decreased by 14 %,IM 0.2 μmol/L showed 40 % reduction,and combination group were significantly depressed for the mRNA expression by 60 %. Conclusion The combination of DAC and IM showed synergistic effects on cell death in K562 cells. These data suggested that DAC used in combination with IM has clinical potential in the treatment of chronic myeloid leukemia.

Objective To study the normal values of blood pressure in healthy full-term infants in the first 7 days of life,and to determine the critical hypertension and hypotension in newborn infants.Methods Omni-Trak~(TM)NVS monitor with DINAMAP was used to measure the blood pressure.Systolic,diastolic and mean blood pressure were measured every day by DLNAMAP in a cohort of 50 healthy full-term infants in the first 7 days of life.Results 1.Systolic,diastolic and mean blood pressure of healthy full-term infants were significantly influenced with the age of birth.2.Stepwise linear multiple regressions was used to examine the multiple correlations among days of life,birth weight and gestational age.3.Optimal regression equations were built respectively.SBP=32.4+0.7X_1+(3.7X_2+)(0.6)X_3(mm Hg).DBP=13.1+ 0.6X_1+2.9X_2+0.6X_3(mm Hg).MAP=17.3+0.4X_1+2.4X_2+0.8X_3(mm Hg).(3.MAP=)(DBP+)(0.45)(SBP-DBP)(mm Hg).4.The critical hypertension and hypotension in newborn infants were obtained.Conclusions (Du)ring the first 7 days of life,there is a progressive rise in blood pressure of healthy full-term infants.There are linear reliance correlations between systolic,diastolic,mean blood pressure and day of life,birth weight gestational age.The critical hypertension and hypotension in newborn infants is important for clinic diagnosis.

As a novel transdermal drug delivery technology of minimally invasive, safe and efficient, microneedles have received increasing attention. The microchannels formation by microneedles onto the skin is a prerequisite and key for microneedles to deliver drugs. However, there is still a lack of systematic evaluation in skin microchannels. This review summarized influencing factors and evaluation methods in microchannels formation and healing by microneedles, including geometric parameters, materials for preparation, drugs, penetration parameters, differences among the skin of subjects, and presence or absence of occlusion. This review provides reference for other scholars to further study the effectiveness and security of microneedle applications.

Objective To judge method performance of routine biochemistry parameters on Roche Modular PPI testing system by using westgard's method evaluation decision chart.Methods We assessed imprecision(CV)from internal quality control and inaccuracy(bias)from external quality control evaluation.Combined estimates of imprecision and inaccuracy by plotting imprecision as the x-coordinate and inaccuracy as the y-coordinate to locate an expected operating point of every item on the chart.By comparing this operating point with allowable total errors(TEa),we can decide whether the performance is acceptable or not.Results In the 27 different parameters tested,imprecision and bias of calcium were 0.08 mmol/L and 0.06 mmol/L respectively,its performance was marginal.The imprecision of creatinine,urea,glucose, sodium,chloride and phosphorus were 3.20%,2.13%,1.52%,0.89 mmol/L,1.10% and 1.55%,the bias were 4.79%,0.96%,4.63%,0.80 mmol/L,1.74% and 4.13% respectively,their performance was good.M1 other 20 items were of excellence performance.Conclusions Routine biochemistry parameters on Roche Modular PPI testing system possessed good precision and accuracy,and their performance were acceptable.To judge method performance of biochemistry testing system by using westgard' s method evaluation decision chart was easy to do and suited for clinical laboratory.

The effect of 1-deoxynojirimycin(DNJ)on the proliferation of rat mesangial cells was observed and its mechanism was explored.The results showed that DNJ significantly inhibited the proliferation of rat mesangial cells induced by high glucose in time-and dose-dependent manners.DNJ significantly decreased expressions of?-smooth muscle action(?-SMA),integrin?1 mRNA and protein and focal adhesion kinase (FAK)protein stimulated by high glucose in rat mesangial cells(P

Objective To observe the changes of brain heme oxygenase-1/carbon monoxide (HO-1/CO)in neonatal rats with hypoxic-ischemic brain damage(HIBD)injury and investigate the role of HO-1/CO in the recovery of HIBD. Methods Eighteen 7-day-old Wistar rats were randomly divided into sham-operated group,HIBDgroup and HIBDwith zinc protoporphyrin(ZnPP)treatment group (n=6).In the latter two groups,HIBD model was established by unilateral carotid ligation followed by timed exposure to 8%oxygen. Real-time fluorescent quantitative PCR Was performed to determine the expression of HO-1 mRNA and thiobarbituric acid(TBA)method was used to assay malondialdehyde (MDA)contentinthe braintissue of the rats.The cell apoptosis in the brain aRer HIBD was analyzed using flow cytometry,and the blood CO concentration was detected by the absorbance at 420mn and 432 nm.Results Compared to the sham-operated group.HO-1 mRNA expression and blood CO concentration were significantly increased in HIBD group and ZnPP group (P＜0.05).The rats with ZnPP group had significantly lower HO-1 mRNA expression and blood CO concentration than those in HIBD group(P＜0.05).HIBD resultedin significantly increased MDA content and cell apoptosis rate in the rat brain as compared to those in the sham-operated group(p＜0.05),and ZnPP treatment further increased the MDA content and cell apoptosis(P＜0.05). Conclusions Increased brain HO-1 mRNA expression and blood CO concentration in neonatal rats with HIBD are probably associated with the spontaneous recovery of neural tissue injury.

Objective: To evaluate the antidiabetic effect of Callicarpa nudiflora extract in streptozotocin-induced diabetic rats. Methods: The chemical constituents in Callicarpa nudiflora extract were identified by UPLC-Q-TOF-MS. Callicarpa nudiflora extract (0.15 and 0.3 g/kg) was orally administered to streptozotocin-induced diabetic rats for 42 d. The effects of Callicarpa nudiflora extract on body weight, blood glucose, serum insulin, total cholesterol, triglyceride, LDL-C and HDL-C were investigated, and its effect on liver and pancreatic pathology was assessed by histopathological analysis. Moreover, the expression levels of adenosine 5'-monophosphate-activated protein kinase (AMPK), glucose transporter type 4 (GLUT4), phospho-AMPK/AMPK, and p-acetyl-coA carboxylase (P-ACC)/ACC in the skeletal muscles and liver were determined by reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemistry. Results: A total of 34 compounds, including 8 iridoids, 14 phenylpropanoids, and 12 flavonoids, were identified from Callicarpa nudiflora extract. Callicarpa nudiflora extract significantly reduced blood glucose and significantly restored all other biochemical parameters to near normal levels, including serum insulin, total cholesterol, triglyceride, LDL-C, and HDL-C. Callicarpa nudiflora extract improved insulin resistance and reversed the damage in the liver and pancreas caused by diabetes. Furthermore, Callicarpa nudiflora extract increased the expression levels of phospho-AMPK, GLUT4, P-ACC, and insulin receptor substrate-1 and decreased the expression level of PPAR毭 in diabetic rats.Conclusions: Callicarpa nudiflora extract improved oral glucose tolerance, lipid metabolism, insulin resistance, and reversed the diabetes-related damage in the liver and pancreas by activating the AMPK-ACC pathway.

OBJECTIVETo investigate the possible association of IVS5-5G>A polymorphism, positioned in the upstream region of exon 5 of PINK1 gene with the risk for sporadic late onset Parkinson disease (LOPD) in Chinese.

METHODSIntronic regulatory sequence analysis was performed using the web-based in-silico analysis. The authors performed an association study using a case-control series (comprising 382 LOPD patients and 336 controls, Chinese of Han ancestry). Genotyping was performed by PCR-based denaturing high performance liquid chromatography (DHPLC) combined with sequencing analyses. Allele and genotype frequencies were compared by the Chi-square test.

RESULTSIn-silico analysis showed that the intronic IVS5-5G>A polymorphism was located within acceptor site of exon 5 and may be the functional single polymorphism (SNP) in the regulatory region with impact on the splicing of PINK1 gene. Those result yielded statistical significant evidence for the association of PINK1 IVS5-5G>A polymorphism with risk for typical PD in Chinese Han population (OR=1.95, 95%CI: 1.29-2.94, P=0.0012). Homozygote of A allele may have increased risk for LOPD (OR=2.45, 95%CI: 1.27-4.72, P=0.009).

CONCLUSIONThe authors provide the first evidence that the common genetic variation PINK1 IVS5-5G>A may contribute to the risk of LOPD in Chinese population.

Age Distribution ; Age of Onset ; Aged ; Aged, 80 and over ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Exons ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Parkinson Disease ; genetics ; Polymorphism, Genetic ; Protein Kinases ; genetics ; Sex Distribution

We have constituted a mouse model for fetal blood transplantation (FBT) to cross over major histocompatibility complex (MHC) without causing serious GVHD. It seems that full matching at the MHC appears not necessary for FBT, while the nucleated cell dose is critical. Two fetal blood units were combined from different donors to increase the stem/progenitor cell dose so as to explore the possibility of MHC-mismatched allogeneic transplantation. 26 out of 40 mice in mixed FBT group survived in the observation period of 60 days after transplantation without obvious GVHD. Double chimerism was demonstrated by PCR and flow cytometric analysis; and skin transplantation test proved the induction of donor specific immune tolerance. Our data suggest that two MHC-mismatched allogeneic donor fetal blood units could simultaneously engraft and reconstitute immune and hematopoietic system in a mouse model. The result may be beneficial for the expansion of cord blood application and enables more patients to share the advantages of cord blood transplantation.
Animals ; DNA ; biosynthesis ; Female ; Fetal Blood ; immunology ; transplantation ; Graft vs Host Disease ; immunology ; mortality ; H-2 Antigens ; immunology ; Hematopoiesis ; immunology ; Hematopoietic Stem Cell Transplantation ; methods ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Survival Rate ; Transplantation Chimera ; genetics ; immunology ; Transplantation Tolerance ; immunology