OBJECTIVE:To determine serum concentrations of western medicines in patients treated with "traditional Chinese antiepileptic medicine" alone and to evaluate the curative effects.METHODS:A total of 60 epileptic patients who visited our hospital between Feb.1997 and June 2006 were subjected to plasma drug level monitoring and during which the patients were treated with "traditional Chinese antiepileptic drugs" alone.Plasma concentrations of 4 kinds of western medicin-es were determined by FPIA.RESULTS:Of the 60 cases,valproic acid,carbamazepine,phenytoin and phenobarbitone were detected in 18,40,41,and 47 cases/times,respectively.On average,more than two kinds of western medicines were detected in every patient,and the blood concentrations were mostly beyond effective plasma drug concentration.The total curative effects were unsatisfactory.CONCLUSION:The fact that western medicine ingredients detected in these traditional Chinese antiepileptic medicines is inconformity with medication principle of epilepsy.Traditional Chinese antiepileptic medicines should be used with caution in the clinic in the treatment of epileptic patients.

Objective To explore the expression of methyl CpG binding protein 2 (MeCP2) in gastric cancer tissues and its role in multi-drug resistance (MDR) in gastric cancer cells.Methods The expression of MeCP2 in 90 gastric cancer tissues and the adjacent normal tissues was detected by immunohistochemistry,and the relationship between MeCP2 expression level and clinicopathological parameters was analyzed.The expression level of MeCP2 in gastric cancer cell line SGC7901,MDR cell variants SGC7901/doxorubicin hydrochloride(ADR) and SGC7901/vincristine(VCR)was determined by Western blot.After the expression of MeCP2 was silenced by short interference RNA (siRNA),half inhibitory concentration (IC50) of 5-fluorouraeil and cisplatin in gastric cancer cell was tested by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide(MTT) assay.The t test or chi square test was performed for statistical analysis.Results The positive rate of MeCP2 expression in gastric cancer tissues was 72.2％ (65/90),which was significantly lower than that in adjacent normal tissues (93.3％,84/ 90),and the difference was statistically significant (x2 =14.068,P＜0.01).MeCP2 expression was not correlated with age,tumor maximum diameter and lymph node metastasis (all P＞0.05),however which was related to gender,clinical TNM stages and distant metastasis (x2=4.680,4.186 and 4.327;aH P＜ 0.05).The gray scale ratios of MeCP2/β-actin in SGC7901/ADR and SGC7901/VCR were 0.593 7 ± 0.030 5 and 0.651 2 ± 0.018 6,which were lower than that of parental SGC7901 cells (1),and the differences were statistically significant (t =23.080,17.360;both P ＜ 0.01).After the expression of MeCP2 was silenced by siRNA,the IC50 of 5-fluorouracil and cisplatin in SGC7901 transfected with MeCP2 specific siRNA were (11.540 0±0.469 3) μg/mL and (2.273 0±0.265 4) μg/mL,which were higher than the IC50 of 5-fluorouracil and cisplatin in SGC7901 transfected with non-related oligonucleotide sequence ((8.663 0±0.160 1) μg/mL and (0.884 0 ±0.038 6) μg/mL),respectively.The differences were statistically significant (t =15.380 and 8.153;both P ＜ 0.01).Conclusions The expression of MeCP2 in gastric cancer tissues significantly decreased,which was correlated with clinical stages,distant metastasis.MeCP2 can inhibit the MDR of gastric cancer cell,which indicated the dysregulated expression of MeCP2 might participate in the genesis and development of gastric cancer.

Ginsenosides,which belongs to triterpenoid saponins of plant terpenoids,are the main active components of the valuable medicinal herbs ginseng and American ginseng.Recent studies show that ginsenosides have a variety of beneficial effects,including anti-inflamatory,antioxidant,and anticancer effects.Ginsenosides are synthesized by complicated modification of triterpenoid skeleton after cyclization of 2,3-oxidosqualene through triterpene saponin synthesis pathway in which they share common early steps with phytosterols synthesis.It outlines recent advances for the biosynthesis of ginsenosides,the cloning and characterization of genes encoding key enzymes in the pathway and the basal framework of ginsenosides biosynthesis pathway.The prospects of secondary metabolism engineering in the biosynthesis of plant natural products and its application in ginsenosides biosynthesis are also discussed.

Objective To identify the activity of HCV IRES translation differences and identify the relationship between HCV IRES translation activity and ROS in different concentrations of ferric ammonium citrate ( FAC) in-duction.Methods 1 ) Expression plasmid pCI-Rluc-HCV IRES-Fluc was confirmed by endonuclease digestion as well as luciferase transient expression in Huh-7 cell;2) Controlled by dual-luciferase reporter assay, the differ-ent translation activity of HCV internal ribosomal entry site ( IRES ) was examined in a concentration of 50 μmol/L and 300μmol/L of FAC induction;ROS fluorescent staining method was used to detect the activity of ROS in Huh-7 cells, Western blot method was used to detect the protein expression changes of Nrf2 in Huh-7 cells;3) On the basis of the above experiments, 100 μmol/L DPI was added in 300 μmol/L FAC experimental group, to analyse the changes of HCV replication and ROS production after joining DPI.Results The generation of ROS and the activity of luciferase in the model group were significantly higher than that in the control group ( P<0.05 ) .FAC can enhance the expression of HCV IRES and increase the production of ROS , then causing Nrf2 expression in Huh-7 cell.However,after adding ROS inhibitor DPI, the above functions in Huh-7 cell were weakened.Conclusions The increase of HCV IRES expression induced by FAC is related to excessive ROS pro-duction induced by FAC in Huh-7 cells.

Objective To investigate the role of spo0A gene in growth and sporulation of Clostridium difficile clinical isolates. Methods ClosTron gene knock-out system was used to knock out the spo0A gene of C. difficile strain C25. Bacterial growth curve was plotted by measuring D600 with spectrophotometer in different phases of bacterial growth. Malachite green staining technique was used to count the number of vegetative cells and spores under optical microscope. The sporulation rate was calculated. Results The spo0A mutant and its C25 parental strain showed similar patterns of growth. However, after knock-out of spo0A gene, an asporogenous phenotype was built, while the parental strain could produce spores as usual.Conclusions The spo0A gene plays a key role in sporulation but not growth of C. difficile strain.

In the bulge region of the hair follicle, a densely and concentrically packed cell mass is encircled by the arrector pili muscle (APM), which offers a specilized microenvironment (niche) for housing heterogeneous adult stem cells. However, the detailed histological architecture and the cellular composition of the bulge region warrants intensive study and may have implications for the regulation of hair follicle growth regulation. This study was designed to define the gene-expression profiles of putative stem cells and lineage-specific precursors in the mid-portions of plucked hair follicles prepared according to the presence of detectable autofluorescence. The structure was also characterized by using a consecutive sectioning technique. The bulge region of the hair follicle with autofluorescence was precisely excised by employing a micro-dissection procedure. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to identify the gene expression profiles specific for epithelial, melanocyte and stromal stem cells in the bulge region of the hair follicle visualized by autofluorescence. The morphology and its age-dependent changes of bulge region of the hair follicles with autofluorescence segment were also examined in 9 scalp skin specimens collected from patients aged 30 weeks to 75 years, by serial sectioning and immuno-staining. Gene expression profile analysis revealed that there were cells with mRNA transcripts of Dct(Hi)Tyrase(Lo)-Tyrp1(Lo)MC1R(Lo)MITF(Lo)/K15(Hi)/NPNT(Hi) in the bulge region of the hair follicle with autofluorescence segments, which differed from the patterns in hair bulbs. Small cell-protrusions that sprouted from the outer root sheath (ORS) were clearly observed at the APM inserting level in serial sections of hair follicles by immunohistological staining, which were characteristically replete with K15+/K19+expressing cells. Likewise, the muscle bundles of APM positive for smooth muscle actin intimately encircled these cell-protrusions, and the occurrence frequency of the cell-protrusions was increased in fetal scalp skin compared with adult scalp skin. This study provided the evidence that the cell-protrusions occurring at the ORS relative to the APM insertion are more likely to be characteristic of the visible niches that are filled with abundant stem cells. The occurrence frequency of these cell-protrusions was significantly increased in fetal scalp skin samples (128%) as compared with the scalp skins of younger (49.4%) and older (25.4%) adults (P<0.01), but difference in the frequency between the two adult groups were not significant. These results indicated that these cell-protrusions function as a niche house for the myriad stem cells and/or precursors to meet the needs of the development of hair follicles in an embryo. The micro-dissection used in this study was simple and reliable in excising the bulge region of the hair follicle with autofluorescence segments dependent on their autofluorescence is of value for the study of stem cell culture.

Objective: Investigate the clinicopathological features for secretory carcinoma of breast (SCB). Methods: The clinical data of 3 SCB cases were collected, immunohistochemical staining was performed by the streptavidin-peroxidase (SP) method to test the expression of the antibodies: ER, PR, HER-2, Ki-67, S100, CK5/6, p63, SMA, calponin, GCDFP-15, and EGFR. Fluorescence in situ hybridization (FISH) was used to detect the ETV6-NTRK3 gene fusion. Results: ER was focal weakly positive in case 1 and case 2 (about 5%) , and negative in case 3. PR was focal weakly positive in case 1 (about 5%) and completely negative in case 2 and case 3. Three cases showed that HER-2, SMA, calponin, GCDFP-15 were negative, while S100, CK5/6, EGFR were diffuse strongly positive. The proliferation index was nearly 15% in case 1 and case 2, and 10% in case 3. p63 was negative in mostly tumor cells of case 1, and focal positive expression in the nucleus and cytoplasm. In case 2, p63 was completely negative. However, p63 was observed positive in the cytoplasm as well as some secretory material in case 3. ETV6-NTRK3 gene fusion detection by FISH was positive in all cases. Conclusions SCB is a rare low grade triple-negative breast cancer with the unique pathomorphologic features, while its recurrent t (12; 15) (p13; q25) translocation resulting in ETV6 -NTRK3 gene fusion. It has the indolent clinical behavior and good prognosis.

OBJECTIVETo study the role of mast cells in the differential diagnosis of cellular leiomyoma and endometrial stromal sarcoma of uterus and its mechanism.

METHODSUsing SP immunohistochemical technique, the expression of proliferating cell nuclear antigen (PCNA) and mast cells in 25 cellular leiomyoma (CL) and 26 endometrial stromal sarcoma (ESS) of uterus were examined. The expression of estrogen receptor (ER) and CD44v3 in cellular leiomyoma was also studied.

RESULTSThe expression of PCNA was not significantly different from CL or ESS (P > 0.05), while mast cell count was statistically different between them (P < 0.01). Using a value of less than 7 mast cells per high power field was useful for the diagnosis of ESS, yielding 100% sensitivity and 92.0% specificity. There was a positive correlation between the mast cell count and CD44v3 in CL (r(s) = 0.589, P < 0.01), though no correlation was observed between mast cell count and PCNA or ER.

CONCLUSIONNumber of mast cells is valuable for the discrimination of CL from ESS in the uterus. The mechanism and the role of higher quantity of mast cells in CL need further study.

Adult ; Aged ; Female ; Humans ; Hyaluronan Receptors ; analysis ; Leiomyoma ; chemistry ; pathology ; Mast Cells ; pathology ; Middle Aged ; Proliferating Cell Nuclear Antigen ; analysis ; Receptors, Estrogen ; analysis ; Sarcoma, Endometrial Stromal ; chemistry ; pathology ; Uterine Neoplasms ; chemistry ; pathology

The mechanism of injury on the human glomerular endothelial cells (ciGENC) induced by preeclampsia serum was investigated. Concentration of maternal serum sFlt-1 protein was detected by ELISA. Fluorescently-labeled bovine serum albumin infiltrating through lower chamber of Transwell was measured by multifunction microplate reader. Morphologic change of ciGENC was observed under inverted phase contrast microscope. The concentration of sflt-1 in preeclampsia groups was significantly increased as compared with control group (P<0.01). Permeability in preeclampsia groups was significantly increased as compared with control group (P<0.01). By contrast with severe preeclampsia group, the permeability of ciGENC monolayer in mild preeclampsia group was decreased significantly (P<0.05). Intervention of exogenous VEGF significantly decreased permeability of ciGENC in preeclampsia groups. It was concluded that sFlt-1 increased ciGENC permeability by damaging integrity of endothelial barrier function.

This study examined the effect of over-expression of sFlt-1 by trophoblasts on the barrier function of glomerular endothelial cells and the role of VEGF in this process in order to explore the pathogenesis of glomerular disease in preeclampsia. SFlt-1 expression in the human trophoblasts (TEV-1 cells) was enhanced by transfecting sFlt-1 plasmid DNA into TEV-1 cells. The monolayer barrier function of glomerular endothelial cells (ciGEnCs) was determined by measuring the fluorescence intensity of bovine serum albumin (BSA) that crossed the monolayer of glomerular endothelial cells. The results showed that the over-expression of sFlt-1 by TEV-1 cells led to the barrier dysfunction of ciGEnCs, and the exogenous VEGF could alleviate the ciGEnCs dysfunction resulting from the over-expression of sFlt-1 to a certain extent. It was concluded that the dysregulation of sFlt-1 and VEGF in preeclamptic pregnancy may contribute to the barrier dysfunction of glomerular endothelial cells, and VEGF may play an important role in maintaining the barrier function of glomerular endothelial cells, but it may not be the sole factor.