BACKGROUND:Tetramethylpyrazine (TMP) can inhibit the expression of vascular endothelial growth factor (VEGF), but it is uncertain that TMP inhibit the growth and proliferation of HL-60 leukemic cells induced by VEGF.OBJECTIVE:To observe the effect of TMP on the proliferation of HL-60 leukemic cells induced by VEGF.DESIGN:Repetitive measurement and observation.SETTING:School of Medicine, Wuhan University of Science and Technology.MATERIALS:The experiment was carried out in the Molecular Biology Laboratory Center, School of Medicine, Wuhan University of Science and Technology from March to June in 2007. Human leukemic cell line HL-60 cells were purchased from Shanghai Institute of Cell Biology. TMP hydrochloride injection was produced by Wuxi Seventh Pharmaceutical Products Limited (Lot number:011014), protamine sulfate injection was produced by Shanghai First Biochemical Pharmaceuticals (Batch number:010302), and immunohistochemistry kit was purchased from Boster company.METHODS:①Human leukemic cell line HL-60 cells at log phase were used for the experiments. Cells were treated with 100 μg/L VEGF, and then TMP at final concentrations of 1.5, 15, 150 mg/L was added into culture medium. While the cells in medium without TMP were taken as blank control group, and the cells in medium with 20 mg/L protamine as positive control group. Meanwhile cells without treatment of VEGF were served as VEGF control group. After cells were incubated for 48 hours, the growth inhibiting rate of HL-60 cells was detected by MTT assay.②After HL-60 cells were treated with TMP at the final concentrations of 1.5, 15, 150 mg/L for 24 hours, the protein expression of VEGF in HL-60 cells was examined by SP immunohistochemistry.MAIN OUTCOME MEASURES:①Growth inhibiting rate of HL-60 cells.②Protein expression of VEGF.RESULTS:①Growth inhibiting rate of HL-60 cells:After HL-60 cells induced by VEGF were treated with 15 and 150 mg/L TMP, the absorbance value was significantly lower than that in VEGF control group (P < 0.05).②Protein expression of VEGF:After HL-60 cells were treated with TMP for 24 hours, the protein expression of VEGF was down-regulated with increasing TMP concentration in a dependent manner. Significant differences were observed in the protein expression of VEGF between cells treated by TMP and the controls (P < 0.01).CONCLUSION:TMP can inhibit the proliferation of HL-60 cells stimulated by VEGF and down-regulate the expression of VEGF.

Objective To observe the morphological changes of hairs in two patients with loose anagen hair syndrome.Methods Light microscopy,scanning electron microscopy and transmission electron microscopy were used to observe the morphology of hair shaft and follicles from two patients,including a 3-year-old girl child and her mother,with loose anagen hair syndrome.Results Light microscopy revealed that hair bulb was deformed,hair shaft was distorted,damaged,and even disrupted,and hairs tapered in diameter at their distal end.Scanning electron microscopy showed deformed or distorted hair shaft and wave-like edge of hair cuticles.Transmission electron microscopy revealed that pathological changes were mainly localized in the inner root sheath with vacuolization in both inner and outer root sheath cells.Intercellular adhesion was weak with a decrease or disappearance of desmosomes.Conclusion The pathological changes of hairs are mainly localized in the inner root sheath in patients with loose anagen hair syndrome.

Objective To screen differentially expressed microRNAs at different stages of hair cycle in a murine model.Methods This study included 30 inbred female C57BL/6 mice (age,6-8 weeks; body weight,15-18 grams).Hair growth cycle was induced in the back skin of C57BL/6 mice by application of wax/rosin followed by depilation under anesthesia witl 1％ chloral hydrate.Three mice were sacrificed by cervical dislocation on day 0,8 and 20 after the induction,and skin tissue was achieved from the same depilated areas parallel to the spine.Total RNAs were extracted from the murine skin and subjected to microarray analysis of microRNA expression.Results Compared with telogen skin,the murine anagen skin showed a higher expression of miR-690,obselote-49 and miR-1308,but a lower expression of miR-291a-5p and miR-212.The expressions of miR-690,obselote-49 and miR-31 were significantly up-regulated,while those of miR-127-3p and miR-212 were downregulated in the catagen skin in comparison with the telogen skin.Conclusion Seven microRNAs are identified in this study to be differentially expressed in murine skin between different stages of hair cycle,which may provide a direction for future research.

Objective To investigate the effect of Ginsensode Rgl on the expression of Neurogranin (Ng) and behavioral alteration in cortex and hippocampus of rats with chronic stress model.Methods A total of 36 adult male SD rats were randomly divided into control group (CON),model group (CUS) and treatment group (CUS-G).The chronic stress model was established by chronic unpredictable stress.The Morris water maze was used to study the learning and memory ability.The content of Ng in cortex,hippocampus was detected by RT-PCR and Western blot.Results The water maze test showed that after chronic stress,animal learning and memory ability decreased significantly,while the treatment group rats escape latency was significantly reduced (P＜0.05);after 6 weeks of stress,the cortex and hippocampus Ng mRNA levelschronic stress rats were markedly lower than that of model rats respectively (P＜0.05,P＜0.01,P＜0.05).The cerebral cortex and hippocampus Ng mRNA levels in treatment group were significantly increased compared with that of model group respectively (P＜0.01,P＜ 0.05,P＜0.05);The cerebral cortex and hippocampus Ng levels of chronic stress rat were significantly decreased when compared with that of the model rats respectively (P＜0.05,P＜0.01,P＜0.05),The cerebral cortex and hippocampus Ng content were significantly increased in treatment group compared with the model group respectively (P＜0.01,P＜0.05).Conclusions Chronic stress can change the behaviors of nice in recognization and memory The contents of Ng and the supplement of Ginsensode Rg1 have positive adjustment.

Objective To explore the effect of prevention and treatment of external application of Algoplaque for controlling phlebitis caused by peripheral indewelling needle in vein for patients. Methods This research was divided into two parts,prevention and treatment. As for prevention research,patients were randomly divided into the experimental and the control groups,each group included 30 patients. In the experimental group,we applied directly external application of Algoplaque at the upper of needle puncture site of the vein and nearby the eye. In the control group,we applied the film directly to fix the indwelling needle. As for the treatment research, it was carried out in patients with occurred phlebitis, who were randomly divided into two groups,the experimental group included 30 cases of patients and the control group included 28 cases of patients. Observation time was one to five days. Results The incidence of phlebitis in the experimental group of prevention research was 23%, in the control group it was 90%. The incidence of phlebitis in the experimental group was significantly lower than that of the control group. The effective rate in the experimental group of treatment research was 96.7% and it was 67.9% in the control group. The difference was very significant. Conclusions External application of Algoplaque can effectively control phlebitis caused by peripheral indewelling needle in vein.

Objective To investigate the mutation of type Ⅱ human hair basic keratin (hHb) gene in a family with monilethrix.Methods Scanning electron microscopy was used to observe the structure of hair shafts.With informed consent,blood samples were drawn from affected and unaffected membets in this family,as well as from 50 healthy controls.Genomic DNA was isolated from these samples.The exon 1 and exon 7 of hHb1,hHb3 and hHb6 were amplified by polymerase chain reaction (PCR).All the PCR products were sequenced directly using ABI3730 automated sequencer.DNA sequence alignment was carried out with BLAST software.Results A typical beaded appearance was observed in affected hairs by using scanning electron microscopy.There were obvious longitudinal ridges and sulcuses in hair node.and hair cuticles were irregularly shaped.Most cortex and medullary substance were absent in affected hairs of a patient.After sequence alignment,a G1289A point mutation in exon 7 of hHb6 gene,which led to a substitution of arginine for glutamide at codon 430,was detected in affected members of this family,but not in unaffected family members or 50 unrelated human controls.No mutation was observed in exon 1 or exon 7 of hHb1 and hHb3 gene or exon 1 of hHb6 gene.Conclusion The missense mutation of R430Q is a novel mutation.which may be associated with the pathogenesis of monilethrix in this pedigree.

Grounded theory was used to summarize and analyze influencing factors and their mechanism on availability of essential medicine.Four factors which influenced the availability of essential medicine were singled out:defective top design in the essential medicine system,interactions among its policies,deviations in the policy implementation by government agencies,and deficiency of supporting policies for the system.The availability of essential medicine in rural areas was influenced by a variety of factors.The ideas and methods of the grounded theory prove helpful for this study.In the future studies,both qualitative and quantitative study should be made to perfect this model formed by the grounded theory,to identify roadblocks and underlying causes in order to provide evidence for improving availability of essential medicine in rural areas.

Objective To study the different types of medication guidance for community doctors in the context of essential medicine system,and analyze its influencing factors.Methods SPSS 1 9.0 was called into play for statistical analysis of data gathered,and the medication guidance behavior of the doctors were graded on the two dimensions of personal willingness and practical action.With important quadrant method,the medication guidance behavior of such doctors was classified,while the multiple linear regression was used to study the influencing factors.Results Most of the community doctors of the fourteen counties fall into the area of “high willingness and less action”,with only a few falling into the area of the “high willingness and more action”. Major influencing factors of their willingness on medication guide were found to include impacts on job autonomy,attention to essential medicine system, and the forms to improve income.Major factors of the doctors'actions on medication guide include awareness of essential medicines,awareness of essential medicines catalog,and training outcomes. Conclusion The community doctors have high personal willingness for medication guidance related to essential medicines,but only a number of them carried it in their work.Therefore,it is necessary to take actions to strengthen awareness of the doctors for medication guidance and encourage them to carry it out.

Objective To identify the impact factors for community doctors with the essential medicine system in place,in support of mobilizing them and successful implementation of the essential medicine system in the community.Methods Based on the theory of ERG hierarchy of needs,the pathway analysis method was called into play to identify the three factors affecting community doctors,namely personal survival,interpersonal relations and self-development.Results Major factor of which is the interpersonal factor,with a score of 0.549,while the latter two factors do not directly affect the survival and development.The two however work on each other,and work on the interpersonal factor as well,with the impact coefficients of 0.320 and 0.402 respectively.Conclusion With the essential medicine system in place,the largest impact factor for community doctors'enthusiasm is the interpersonal one,involving the doctor-patient communications among others.This serving as the breakthrough point may improve the enthusiasm of the community doctors,paving the way for the system in the communities.

Objective To study the inhibitory action of docetaxel (DOC) on the proliferation of HeLa and SiHa cells. Methods Cell morphological changes were observed with inverted phase contrast microscope. MTT was adopted to test and calculate the cell inhibition ratio. Flow cytometry was used to detect cell cycle. Results DOC had an obvious concentration-dependent inhibitory effect on the proliferation of both HeLa and SiHa cells. The inhibition ratio of DOC on SiHa was significantly higher than that on HeLa (P<0.05). DOC blocked HeLa at G2/M phase. Under the effect of DOC, the cell cycle of SiHa was not changed much. Conclusion DOC has an obvious inhibitory action on both HeLa and SiHa cells, which shows a promising prospect of DOC in clinical treatment of cervical cancer.