OBJECTIVE:To investigate the cognition,evaluation and demands of medical staff to the clinical pharmacy work in the secondary and tertiary medical and health institutions in Xi’an city,and provide reference for further promoting the develop-ment of the local clinical pharmacy work. METHODS:20 secondary and tertiary medical and health institutions in Xi’an city were randomly selected to conduct a random sampling questionnaire for physicians,nurses (senior nurses),pharmacists (non-clinical pharmacist)and other medical technicians. And the results were statistically analyzed. RESULTS:Totally 1 020 questionnaires were sent out,851 were effectively received with effective recovery of 83.4%. 45.9% respondents knew clinical pharmacy,and“Col-league”was the main channel;34.3% thought clinical pharmacy“only maintained normal operation. 74.7% surveyed medical staff,92.6% surveyed pharmacists and 70.6% surveyed physicians thought clinical pharmacists“should”take round with the doc-tors and nurses;44.3% respondents showed“dissatisfaction”and“general satisfaction”with the clinical pharmacy work in their hospitals. 48.5% respondents would take the initiative to consult the clinical pharmacist for medication;45.5% respondents partial-ly accepted the drug information provided by clinical pharmacists. There were significant differences in the investigation results in aspects of respondents’understanding level and channel for clinical pharmacy,cognition for development situation of clinical phar-macy,evaluation for clinical pharmacists participating round,demand for consulting the medication,acceptance for drug informa-tion provided by clinical pharmacists,and other items(P<0.05). CONCLUSIONS:The cognition and effect of clinical pharmacy work on medical staff need to be further strengthened,clinical pharmacists should also have solid clinical knowledge and the knowl-edge of medicine to meet the demand of medical staff in different positions,the acceptance of medical staff to the pharmaceutical care needs to be further improved. Clinical pharmacists should earnestly fulfill their job functions through various efforts to promote rational drug use.

We analyzed the epidemiological investigation results of the first case of human infected with avian influenza (H5N6) ,and summarized epidemiological characteristics of this case to provide reference to H 5N6 case screening and investiga‐tion in future works .We retrospectively described and analyzed the investigation progress ,specimen detection results of epi‐demic spot and intimate contact people ,and summarize epidemiological characteristics of this case .Result showed that the first case of human infected with H5N6 in Yunnan was a single case ,and did not showed human‐to‐human transmission .This case had contacts with wild bird and had birds’ market activity before diagnoses ;some environment specimen of birds market near patient’s home has detection results of H5N6 positive .This is a cases of local infection and did not show human‐to‐human transmission .From the investigation ,we could deduced that the transmission route is from birds to human or from birds to en‐vironment and then to human .

Objective:To prepare nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR and to examine its cytotoxicity against EGFR positive tumor cells.Methods:By using molecular cloning strategy,prokaryotic expression construct of pET28a-BI7D12-PE38KDEL was generated which consisted of nanobody 7D12 targeting EGFR in the form of a divalent fused with PE38KDEL,a truncated form of pseudomonas exotoxin A via a flexible peptide(G4S)4,and then transformed into E.coli BL21(DE3).Protein expression was induced by adding IPTG,purified by Ni-affinity column chromatography,and verified by Western blot.The binding capacity of the resulted immunotoxin to EGFR-positive cells A549,HT29,MCF-7 and EGFR-negative cells CEM,Jurkat were determined by flow cytometry assay,and its cytotoxicity against the target cells was examined.Briefly,tumor cells were treated with different dosage of the immunotoxin,and the killing efficacy of BI7D12-PE38KDEL on these cells were assessed by WST-1 assay after 72 hours.Results:The SDS-PAGE and Western blot results showed the recombinant immunotoxin BI7D12-PE38KDEL was successfully prepared,and majority of them was expressed in soluble form.BI7D12-PE38KDEL could selectively bind to EGFR-positive cells of A549,HT29,and MCF-7.More importantly,the immunotoxin exhibited much more significant killing effect on these EGFR positive cells compared to the negative control group of CEM and Jurkat cells(P<0.01).Conclusion:In the current study,the nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR was successfully prepared and exhibited a superior inhibition effect for the growth of EGFR-positive cells.

AIM:To investigate the therapeutic and preventive effects of paeoniflorin ( PF) on APP/PS1 mice, and to explore the possible mechanism. METHODS:Fifteen male 5-month-old APP/PS1 non-dominant mice were chosen as normal control group, 15 male 5-month-old APP/PS1 double transgenic mice were used as model group, and 15 male 5-month-old APP/PS1 double transgenic mice treated with 5 mg/kg PF by intraperitoneal injection were allocated in administation group. The learning and memory ability of the mice in each group was detected by Morris water maze. The apoptosis was assessed by TUNEL fluorescence staining. The protein expression of PI3K, Akt, p-PI3K, p-Akt, caspase-3, caspase-9, Bcl-2 and Bax in cerebral cortex and hippocampus was detected by Western Blot. The protein expression levels and distribution of caspase-3 and caspase-9 were detected by immunohistochemistry. RESULTS:(1) Compared with nor-mal control group, the learning and memory ability declined in APP/PS1 model group. Compared with APP/PS1 model group, PF obviously improve the ability of learning and memory in mice. (2) Compared with normal control group, the ap-optosis of nerve cells in APP/PS1 model group significantly increased and distributed in wider areas, while that in PF group was reduced (P<0.05). (3) Compared with APP/PS1 model group, PF could significantly lower pro-apoptotic factors, caspase-3, caspase-9 and Bax (P<0.05), and increase the expression of anti-apoptotic factors, p-PI3K, p-Akt and Bcl-2 (P<0.05). CONCLUSION:PF can up-regulate the expression of Bcl-2 and down-regulate the expression levels of caspase-9, caspase-3 and Bax via the activation of PI3K/Akt pathway, thereby inhibiting the nerve cell apoptosis and pro-tecting the nerve cells, so as to treat neurodegenerative diseases.

Objective：This study was designed to investigate BCRP gene expression in normal lung and in non-small cell lung cancer tissue. Methods：RNA was extracted immediately from the fresh normal lung tissue and viable tumor tissue harvested from surgically resected specimens of non-small cell lung cancer patients. cDNA of BCRP gene was prepared by RT-PCR and then amplified by PCR. The cDNA products from those specimens were transferred to blotting membrane through electrophoresis and transferring technique. Southern blot hybridization was eventually performed to detect the expression of BCRP gene. Results：RNAs were extracted from 8 tumor tissues and 12 pairs of tumor tissue and normal lung tissue were harvested from the same lung. Four patients’RNA samples with poor quality due to degrading were discarded. cDNA products of BCRP gene were obtained by RT-PCR and amplified by PCR in the remain 16 patients’RNA samples. Through Southern blot hybridization, BCRP gene was found to have slight expression in various amounts in all normal lung tissue(10/10) and only in half of tumor tissue samples (8/16). Conclusion：BCRP gene is slightly expressed in a variable amount in all normal lung tissue and only in half of tumor tissue for this group of patients with non-small cell lung cancer. Therefore chemotherapy with anthracycline may possibly induce the overexpression of BCRP gene and at last to develop multidrug resistance.

Background and objectiveMaintenance of genomic integrity is essential to ensure normal organismal development and to prevent diseases such as cancer. PR-Set7 (also known as Set8) is a cell cycle regulated enzyme that catalyses monomethylation of histone 4 at Lys20 (H4K20me1) to promote chromosome condensation and prevent DNA damage. Recent studies show that CRL4CDT2-mediated ubiquitylation of PR-Set7 leads to its degradation during S phase and atfer DNA damage. hTis might occur to ensure appropriate changes in chromosome structure during the cell cycle or to preserve genome integrity atfer DNA damage.Methods We developed a new model of lung tumor development in mice harboring a conditionally expressed allele of Cul4A. We have therefore used a mouse model to demonstrate for the ifrst time that Cul4A is oncogenicin vivo. With this model, staining of PR-Set7 in the preneoplastic and tumor lesions in AdenoCre-induced mouse lungs was performed. Meanwhile we identiifed higher protein level changes of γ-tubulin and pericentrin by IHC.Results hTe level of PR-Set7 down-regulated in the preneoplastic and adenocarcinomous lesions following over-expression of Cul4A. We also identiifed higher levels of the proteins pericentrin and γ-tubulin in Cul4A mouse lungs induced by AdenoCre.Conclusion PR-Set7 is a direct target of Cul4A for degradation and involved in the formation of lung tumors in the conditional Cul4A transgenic mouse model.

OBJECTIVETo investigate the rate of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphomas (NHL) and its pathogenesis and pathologic significance.

METHODSPCR analysis of monoclonal, polyclonal and dual rearrangements of IgH and TCR gamma, TCR beta genes was carried out in 125 cases of NHL to evaluate the rate of dual rearrangements, immunohistochemistry was performed for a Ki67 protein expression in 117 cases and the proliferation index was calculated. The relationship between antigen receptor gene rearrangements and proliferation index was analyzed.

RESULTSCombination of the two pairs of IgH gene primers with the multiplex PCR for TCR gamma and TCR beta gene revealed dual rearrangements in 8% (8/96) of B-NHL, 17% (5/29) of T-NHL. In B cell NHL, IgH gene monoclonal, dural and polyclonal rearrangements were identified in 65, 8 and 15 cases respectively, while in T-cell NHL, they were in 15, 5 and 9 cases, respectively. There was no significant difference between proliferation index and monoclonal, dual, polyclonal rearrangements in both B-NHL and T-NHL by One-way test.

CONCLUSIONDual rearrangements in NHL are not rare and have no relationship with proliferation index.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carrier Proteins ; genetics ; Cell Proliferation ; Child ; Female ; Gene Rearrangement ; Genes, T-Cell Receptor gamma ; genetics ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Lymphoma, B-Cell ; genetics ; metabolism ; pathology ; Lymphoma, Non-Hodgkin ; genetics ; metabolism ; pathology ; Lymphoma, T-Cell ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Polymerase Chain Reaction ; Receptors, Antigen ; genetics ; Young Adult

Objective:The prokaryotic expression vector of human anti-Siglec-9 Fab fragment antibody was constructed and purified,while was identified. Methods:The variable and conserved regions of heavy chain and light chain were obtained by polymerase chain reaction respectively(PCR),which was combined by overlap extension PCR and was digested with restriction enzyme,and then it was transformed into Escherichia coli BL21 and was purified by His-trap Lambda Fab column and AKTA system. SDS-PAGE,ELISA and Western blot were used for the identification of human anti-Siglec-9 Fab fragment antibody. The effect of human anti-Siglec-9 Fab fragment antibody on regulating the mRNA expression of TNF-α,IL-1,IL-6,IL-8 was detected by real-time PCR. Results:Successfully obtained the chains of heavy and light, while constructed an activation human anti-Siglec-9 Fab fragment antibody which could specifically bind to Siglec-9 protein. The human anti-Siglec-9 Fab fragment antibody could specifically bind to Siglec-9 was confirmed by SDS-PAGE,ELISA and Western blot. The human anti-Siglec-9 Fab fragment antibody inhibited the mRNA expression of TNF-α,IL-1, IL-6,IL-8. Conclusion:Successful prokaryotic expression,purification,character analysis,and suppressed the mRNA expression of in-flammatory cytokines with the human anti-Siglec-9 Fab fragment antibody and lay the biology foundation for the further study.

Objective:To explore influence of amlodipine besylate tablet combined metoprolol on serum inflammatory factor levels and cardiac function in aged patients with essential hypertension(EH)complicated chronic heart failure(CHF).Methods:A total of 98 aged EH+CHF patients treated in our hospital from Jan 2014 to Oct 2015 were se-lected.Patients were randomly and equally divided into metoprolol group(received metoprolol based on routine treatment)and combined treatment group(received amlodipine besylate tablet based on metoprolol group),both groups were treated for six months.Serum levels of interleukin(IL)-1,IL-6 and tumor necrosis factor(TNF)-α, left ventricular end-diastolic dimension(LVEDd),left ventricular end-systolic dimension(LVESd)and left ventric-ular ejection fraction(LVEF)before and six months after treatment,and therapeutic effect were measured and compared between two groups.Results:Compared with before treatment,after six-month treatment,there were significant reductions in serum levels of IL-1,IL-6,TNF-α,LVEDd and LVESd,and significant rise in LVEF in two groups,P=0.001 all;compared with metoprolol group after six-month treatment,there were significant reduc-tions in serum levels of IL-1[(16.21 ± 1.45)ng/L vs.(12.34 ± 1.32)ng/L],IL-6[(46.21 ± 5.32)ng/L vs. (36.42 ± 4.51)ng/L],TNF-α[(23.12 ± 2.14)ng/L vs.(19.25 ± 2.41)ng/L],LVEDd[(51.68 ± 4.74)mm vs. (42.31 ± 4.25)mm]and LVESd[(46.27 ± 4.78)mm vs.(39.06 ± 4.35)mm],and significant rise in LVEF [(43.26 ± 4.37)% vs.(52.95 ± 5.18)%]in combined treatment group,P=0.001 all.Total effective rate of com-bined treatment group was significantly higher than that of metoprolol group(93.88% vs.79.59%),P= 0.037. There was no significant difference in incidence rate of adverse reactions between two groups,P=0.505. Conclu-sion:Amlodipine besylate combined metoprolol possesses significant therapeutic effect on aged EH + CHF patients, which may reduce serum inflammatory factor levels,relieve ventricular remodeling and improve cardiac function.

OBJECTIVETo compare therapeutic effects of red-hot needle therapy and filiform needle therapy on nodules of breast of the type of stagnation of liver-qi and phlegm coagulation.

METHODSSix hundred cases were randomly divided into a treatment group and a control group, 300 cases in each group. The treatment group were treated with red-hot needle pricking at the proliferative parts and Ashi points as main, and the control group with filiform needle therapy.

RESULTSIn the treatment group, 240 cases were cured, 58 cases improved and 2 cases were not cured with an effective rate of 99.3%; and in the control group, 113 cases were cured, 165 cases improved and 22 were not cured with an effective rate of 92.7%, with a significant difference between the two groups (P = 0.001).

CONCLUSIONThe therapeutic effect of red-hot needle therapy is better than that of filiform needle therapy on nodules of breast of the type of stagnation of liver-qi and phlegm coagulation.

Acupuncture Therapy ; methods ; Adult ; Female ; Fibrocystic Breast Disease ; etiology ; therapy ; Humans ; Middle Aged