OBJECTIVE: To explore the management pattern that conducive to the development of the Chinese Herbal Pieces Department.METHODS: The pros and cons of the comprehensive pharmacy management pattern for Chinese Herbal Pieces Department were analyzed.RESULTS: The introduction of the comprehensive pharmacy management pattern in the Chinese Herbal Pieces Department resulted in the reduction of the human,material and financial resources;however,it also led to declining of the quality of the cut crude drug,reduction in drug consumption,lowering of pharmacists' sense of responsibility,presence of more risk factors of dispensing errors,lowering of social concern,weakening of the position of Chinese medicine and marginalization of status of Chinese medicine.CONCLUSION: The inclusion of Chinese Herbal Pieces Department into comprehensive pharmacy management should be carried out with prudence or it might as well to maintain the relative independence of the Chinese Herbal Pieces Department so as to protect and develop the Chinese medicine.

Lack of teachers' participating enthusiasm is the main problem that the institutions of teaching and faculty development face during the promoting process of faculty development projects. In this study, service marketing triangle theory was applied to the project promotion of the institutions of teaching and faculty development, the service marketing triangle theory for the project promotion of the center for teaching and faculty development was established. Through the three aspects of outside marketing, internal marketing and interaction marketing, the promotion strategies of faculty development projects were proposed. The application of these strategies in promoting the faculty development projects will enhance the teachers' initiative and enthusiasm to participate in service projects, so as to push faculty development.

Objective To study the mechanism that glial cell line-derived neurotrophic factor (GDNF)promotes human glio-ma cells proliferation.Methods We divided glioma samples into two groups,including low-grade glioma group and high-grade glio-ma group,while cerebral contusion patients were treated as the control group,12 cases in each group.C6 glioma cell lines were di-vided into three groups,such as GDNF group,BSA(bovine serum albumin)group and control group.CCK-8 (cell counting kit-8) was used to detect the cell proliferation,while Western blot was used to detect the expression of AKT,p-AKT,β-catenin and p-β-catenin in each group.Results Comparing with the control group,the expression levels of AKT,p-AKT,β-catenin and p-β-catenin in glioma group had a significantly increased (P <0.05).Meanwhile,the high-grade gliomas group also had a significant increase in those more than low-grade gliomas group (P <0.05).CCK-8 test showed that the cell proliferation in GDNF group was significant-ly higher than the control group (P <0.05),and the expression levels of p-AKT,β-catenin and p-β-catenin proteins all had a signifi-cant increase (P <0.05).However,the expression level of AKT had no obvious difference.Conclusion GDNF might promote the proliferation of glioma cells by up-regulating the expression of p-AKT,β-catenin and p-β-catenin.

Objective To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. Methods The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral con-tusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to ob-serve the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. Results The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P<0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P<0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expres-sion on 7 d (P<0.05). Conclusion The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.

Objective To analyze nucleophosmin (NPM1) gene mutations in exon 12 in patients with acute myeloid leukemia (AML) and evaluate the clinical appliance of three methods which are frequently used for detecting gene mutation. Methods Genomic DNA from bone marrow of 54 AML patients was detected by PCR for NPM1 exon 12 and screened by PCR-capillary electrophoresis, denature high-performance liquid chromatography (DHPLC) and direct sequencing separately. FLT3-ITD (FMS-like tyrosine kinease internal tandem duplication) was detected by agarose gel electrophoresis and PCR-capillary electrophoresis. Results Seven AML sample harbored NPM1 gene mutations. Five of them were the most common mutation, known as type A (an insertion of a TCTG tetranucleotide at position 960 bp). One of them was type D (an insertion of a CCTG tetranuclectide at position 960 bp). The new variant was a deletion of a TGGCAGTG sequence at 958 bp and insertion of a GCCCGCGGTTTA sequence instead. The detection ratio of the three methods was all 100% and capillary electrophoresis was more rapid, reliable and easier than the other two methods. Moreover it could detect FLT3-ITD simultaneously. The resolving power of DHPLC was affected by many factors. The direct sequencing method was tedious and the heterozygous sequence might be misread. Conclusions There is a new mutation at position 958 bp with a 12-nucleotide insertion and substitution. PCR-capillary electrophoresis is convenient to screen NPM1 mutations of AML in clinical practice.

ObjectiveTo understand the characteristics of mutations in BCR-ABL1 kinase domain mutation,these chronic myeloid leukemia (CML) and Ph positive acute lymphoblastic leukemia (ALL)patients who got imatinib treatment had poor effect.MethodsTotally 177 CML patients and 33 Ph( + )ALL patients were selected at Beijing Dao-Pei Hospital from Sep.2007 to Dec.2010.All of them were Chinese patients.Totally 243 bone marrow or peripheral blood specimens were collected from the patients,who had early effect,then resistance emergenced,or for more than 3 months of poor efficacy.Extracted total RNA from the specimens' nuclear cells,reversed transcription to cDNA.Amplified the whole span of BCRABL1 fusion kinase gene by nest PCR (from 242 to 493 amino acid coding sequence),used the type AB3130XL gene sequencing instrument determinate the gene sequence of ABL1 kinase region and then used the Variant Reporter V1.0 software to analyze the results of gene mutations.ResultsThirty-two kinds of different mutations were detected of ABL1 gene mutations,accounting for 34.2％ (83/243 cases).Among them,the T315I was 12％ (10/83),mutation rate was the highest,followed by Y253H was 11％ (9/83),G250E was 7％ (6/83),E255K was 7％ (6/83),M351T was 6％ (5/83),E459K was 5％ (4/83) ;Q252H,D276G,F317L,E355G,F359V,H396R were all 4％ (3/83).Three cases of insertion mutations were found,including 2 cases of 357-358insk,1 case of V304RfsX17.Seven patients had found existence two or more point mutations.The multiple drug resistance mutations might exist in the same leukemia clone.The same individual was not only contain common resistance mutations,but also rare point mutations,insertion mutations.The mutations might be lead to loss of kinase activity.ConclusionsUnder the imatinib drugs pressure,the ABL1 gene mutation in leukemia cells appears randomly,and results in different resistant clones.Different resistant clones can coexist in the same patients in vivo; resistant clones not only contain point mutations,but also contain inserted deletion mutations.

Objective To develop a multiplex sequence-specific PCR assay for simultaneous screening of 5 types of JAK2 mutations and investigate its clinical application value. Methods Multiplex sequence-specific PCR assay for simultaneous screening of JAK2 V617F, K539L (include 2 types of gene mutations), N542-E543del and E543-D544del mutations were developed. 115 patients with myeloproliferative diseases (MPD) including 61 polycythemia vera (PV) cases, 43 essential thrombocythemia (ET) cases and 11 primary myelofibrosis (MF) cases were analyzed. Results The assay can screen the 5 types of JAK2 mutations efficiently. The detection sensitivity is 1% for JAK2 V617F mutation and 0.1% for the other mutations. JAK2 V617F mutation and JAK2 exon12 mutation were detected in 56 and 3 of the 61 PV samples, respectively. 27 of the 43 ET samples and 6 of the 11 MF samples were JAK2 V617F positive, but no JAK2 exon12 mutation was found in both groups. The 3 cases carrying JAK2 exon12 mutation had the clinical feature of erythrocytosis and erythropoietin-independent erythroid colony formation but without apparent leukocytosis, thrombocytosis and splenectasis. Conclusion The assay can simultaneously screen 5 types of JAK2 mutations with high sensitivity and thus lead to an increased detection rate.

Objective To analyze the etiological factor and genetic feature of a familial hemophagocytic lymphohistiocytosis patient with PRF1 mutation (FHL2) with human herpesvirus 7 (HHV7)infection and its family constellation. Methods Clinical characteristics, laboratory examinations of a FHL2 case with HHV7 infection were reported. HHV1-HHV8 virus DNA was screened by PCR; NK cell function was analyzed by flow cytometry; PRF1 gene mutations were analyzed by PCR and direct sequencing, structure of mutant PRF1 proteins were analyzed using ExPasy and I-TASSER server and genetics pedigree were analyzed. Results The patient's HHV7 viral was detected positive with DNA copy number of 350/106 peripheral nucleated cells. Flow cytometry analysis showed decrease both in proportion of perforin positive NK cells and perforin protein expression. Genetic testing showed PRF1 biallelic heterozygote mutations (c. 503G ＞ A/p. S168N and c. 1177T ＞ C/p. C393R) and pedigree analysis showed they were inherited. The patient was then treated with antivirus therapy, dexamethasone and VP16 therapy, but only achieved partial response. The patient was then followed by human leukocyte antigen 10/10 allele identical nonconsanguinity allogeneic hematopoietic stem cell transplantations (allo-HSCT) and soon the successful implantation of donor hematopoietic cells and persistent recovery was achieved. The patient was now surviving without recurrence for 9 months after allo-HSCT. Conclusions FHL is prone to be misdiagnosed as lymphoma. Genetic analysis of related gene mutation and herpes simplex virus detection will help in early and accurate diagnosis. Allo-HSCT is a fundamental treatment of FHL.

BACKGROUND:Epidural analgesia has been considered a gold standard for postoperative analgesia in the lower limbs. Its outcomes are accurate and adverse reactions are few, so it can be used in the clinic. However, this method has adverse reactions such as hypotension and urine retention. Low molecular weight heparin should be used after operation, which can increase the possibility of epidural hematoma, and limits its application to epidural analgesia in the clinic. At present, few studies concerned ultrasound guided continuous fascia iliaca compartment block technology.
OBJECTIVE:To evaluate the efficacy of postoperative pain relief and the joint rehabilitation between a continuous fascia iliaca compartment block and a continuous epidural analgesia for patients undergoing total hip arthroplasty.
METHODS:A total of 60 patients undergoing a selective total hip arthroplasty were assigned to continuous fascia iliaca compartment block group and continuous epidural analgesia group (n=30). Al patients in both groups received a pre-fluence before general anesthesia. Continuous fascia iliaca compartment block group were injected with 0.25%ropivacaine 30 mL via iliac fascia gap. Continuous epidural analgesia group received 0.20%ropivacaine 10 mL via epidural catheter, indwel ing catheter. When the analgesic effect was identified, anesthesia intubation was carried out. After operation, medicine was given via iliac fascia and epidural analgesia pump in both groups respectively. Postoperative analgesia in single dose was not given. If pain could not be endured, analgesia would be rescued (parecoxib 20-40 mg/time) according to pain degree. Visual analogue scale scores, supplemental analgesia of parecoxib, complication of anesthesia, Harris hip joint scores, day of first walk, and duration of hospital stay were recorded.
RESULTS AND CONCLUSION:No significant difference in visual analogue scale scores, supplemental analgesia, Harris hip joint scores and duration of hospital stay was detected. Day of first walk was earlier in the continuous fascia iliaca compartment block group than in the continuous epidural analgesia group. The complications were apparently lower in the continuous fascia iliaca compartment block group than in the continuous epidural analgesia group. These data indicated that after total hip arthroplasty, two kinds of analgesia methods could provide satisfactory postoperative outcomes. Hip joint was perfectly recovered. However, the complications of continuous fascia iliaca compartment block were less, and helpful to patients’ early off-bed activities, and could be considered as a good choice for analgesia after total hip arthroplasty.

OBJECTIVETo investigate the mechanism of high transcription of the glial cell-line derived neurotrophic factor (gdnf) gene induced by hyperacetylation of histone H3 lysine 9 (H3K9) at its promoter region II in rat C6 glioma cells.

METHODSThe acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and the binding capacity of Egr-1 to its binding site in gdnf promoter were examined by ChIP-PCR in C6 astroglioma cells and normal rat astrocytes, and its changes were investigated in C6 astroglioma cells after treatment with histone acetyltransferase inhibitor curcumin or deacetylase inhibitor trichostatin A.

RESULTSCompared normal astrocytes, C6 astroglioma cells showed significantly increased acetylation level of H3K9 at Egr-1 binding site in gdnf gene promoter region II and Egr-1 binding capacity (P<0.01). Curcumin treatment significantly reduced H3K9 acetylation level at Egr-1 binding site and decreased both the binding of Egr-1 to promoter region II and gdnf mRNA levels in C6 astroglioma cells (P<0.05). Conversely, increased H3K9 acetylation at the Egr-1 binding site induced by trichostatin A significantly increased the binding of Egr-1 to promoter region II and gdnf mRNA expression levels (P<0.05).

CONCLUSIONH3K9 hyperacetylation induces increased Egr-1 binding to gdnf gene promoter II, which might be the reason for the high transcription level of gdnf gene in rat C6 glioma cells.

Acetylation ; Animals ; Astrocytes ; metabolism ; Binding Sites ; Cell Line, Tumor ; Early Growth Response Protein 1 ; metabolism ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; Glioma ; metabolism ; Histones ; chemistry ; Promoter Regions, Genetic ; Protein Processing, Post-Translational ; RNA, Messenger ; Rats ; Transcription, Genetic