Understanding inter-species transmission of influenza viruses is an important research topic. Scientists try to identify and evaluate the functional factors determining the host range of influenza viruses by generating the recombinant viruses through reverse genetics in laboratories, which reveals the viruses' molecular mechanisms of infection and transmission in different species. Therefore, the reverse genetic method is a very important tool for further understanding the biology of influenza viruses and will provide the insight for the prevention and treatment of infections and transmission. However, these recombinant influenza viruses generated in laboratories will become the potential threat to the public health and the environment. In this paper, we discussed the biological safety issues of recombinant influenza viruses and suggested we should set up protocols for risk management on research activities related to recombinant highly pathogenic influenza viruses.
Influenza A Virus, H5N1 Subtype ; genetics ; Laboratories ; Microbiology ; Orthomyxoviridae ; genetics ; Public Health ; Reassortant Viruses ; genetics ; Recombination, Genetic ; Safety

Objective To research the effects of metformin on proliferation and apoptosis in human breast cancer MCF-7 cells and investigate the role of Foxp3 in the effect.Methods The inhibit rate of MCF-7 cells was measured by MTT assay after 48 h metformin treatment(5 mmol/L、10 mmol/L、20 mmol/L).Metformin-induced cell apoptosis was measured by Hoechst 33258 stai-ning and flow cytometry.The expression levels of Foxp3 and caspase-3 mRNA were detected by real-time PCR.Expression of Foxp3 protein was detected by Western blot.Results Metformin markedly inhibited proliferation of MCF-7(P<0.05)compared with control group.And the early apoptotic rates increased to 3.76%,8.96%,and 18.67% with 5 mmol/L、10 mmol/L、20 mmol/L metformin treatment,respectively.Metformin increased caspase-3 mRNA expression levels,and decreased the expression levels of Foxp3 mRNA and protein expression levels(P<0.05,vs.control).Conclusion Metformin could inhibit the proliferation and in-duce apoptosis of MCF-7,and its mechanism maybe related to down-regulated expression of Foxp3.

Skeletal muscle is essential for the glucose disposal in organism, which is one of the major sites of insulin resistance.The over-activity of classical renin-angiotensin system ( RAS) induces the oxidative stress, leads to the impairment of insulin signaling and glucose transport, resulting in insulin resistance of skeleton muscle.The activation of non-classical RAS by inhibiting classical RAS activity, can ameliorate insulin resistance in skeletal muscle tissue.

Hepatitis C virus (HCV), one of the major pathogens of viral hepatitis, causes significant hazards in humans. Interferon treatment in combination with ribavirin is used as the first line clinical treatment for HCV infection. However, good response to this treatment has only been observed in few patients and repeated recurrence has also been reported frequently. Therefore, new antiviral agents and therapies are in urgent demand. Here, we report a newly constructed Escherichia coli RNase P based M1GS ribozyme that can specifically and efficiently target the core gene of HCV. The guide sequence (GS) of this M1IGS was designed according to the sequence of the core coding region of HCV genome. The GS was then covalently linked to the 3' terminus of M1 RNA, the catalytic subunit of RNase P from Escherichia coli. The specification of this sequence-specific ribozyme, M1GS, was then examined using an in vitro cleavage assay. The cytotoxicity and its activity in inhibition of HCV gene expression and viral proliferation were further studied in vivo. Our results show that the reconstructed M1GS ribozyme displayed obvious catalytic activity in cleaving target mRNAs fragment in vitro. Notable reduction in the expression of HCV core protein and a 1 000-fold reduction in viral growth were also observed in cultured HCV infected Huh7.5.1 cells expressing the functional M1GS ribozyme. This study demonstrated a direct evidence for the antiviral activity of the customized M1GS-HCV/C141 ribozyme, and thus provided a promising new strategy for clinical treatment of HCV infection.
Antiviral Agents ; pharmacology ; Escherichia coli ; genetics ; Genetic Engineering ; Hepacivirus ; genetics ; physiology ; RNA, Catalytic ; genetics ; pharmacology ; RNA, Guide ; genetics ; Ribonuclease P ; genetics ; Viral Core Proteins ; genetics

Objective To study the reconstructive treatment of the different types of neurofibroma and neurofibromatosis in different locations. Methods 37 patients with neurobibroma or neurofibromatosis were reviewed, including 8 cases with simple suture after excision, 9 cases with skin grafting in situ, 12 cases with skin flap grafting. The main points of the treating methods for different types of neurofibroma and neurofibromatosis were discussed, and the chief clinical manifestations, therapeutic principle and notices of the operation were then summarized.Results These 37 patients, including 25 cases of neurofibroma, 11 cases of neurofibromatosis typeⅠ and 1 case of neurofibromatosis type Ⅱ, were satisfied with the results after reconstructive operation. Conclusion The methods of reconstructive operation of neurofibroma and neurofibromatosis, such as simple suture after excision, skin grafting in situ, skin flap grafting, the use of skin soft tissue expansion, microskin grafting and so on, must be selected according to the different type and location of the neurofibroma and neurofibromatosis. The reconstructive operation must guarantee the best recovery of the visage and the function on the basis of the most degree of tumor excision.

OBJECTIVE: To provide reference for establishing and improving clinical pharmacists' working mode in women-children special hospital.METHODS: The work carried out by clinical-medical-nursing team and the situation of clinical pharmacists' involvement in clinical treatment were summarized retrospectively.RESULTS & CONCLUSIONS: The medication safety in women-children special hospital has specificity and urgency,and the establishment of clinical-medical-nursing team is an important approach to realize clinical safe and rational drug use.A platform for clinical pharmacists under unified management of organization and system requirement should be set up in hospital to explore a clinical pharmacy working mode with the characteristics of special hospital,carry out clinical pharmacy research and search for quality management and performance management method for clinical pharmacists.

OBJECTIVE To detect AmpC ?-lactamases from isolates of extended spectrum ?-lactamases(ESBLs) positive Escherichia coli and Klebsiella pneumoniae,which were isolated from four hospitals in Hangzhou from 2005 to 2006,and to analyze the antimicrobial activity of clinically commonly used antibiotics in vitro.METHODS Amount of 324 ESBLs positive isolates including E.coli and K.pneumoniae were collected from Zhejiang Provincial People′s Hospital;1st Affiliated Hospital of Zhejiang University;2nd Affiliated Hospital of Zhejiang University and Hangzhou Traditional Chinese Medicine Hospital perspectivly.AmpC ?-lactamase was identified by disc screening testing and three dimensional test,and the genotypes of AmpC ?-lactamase were also determined by multiplex polymerase chain reaction(Multi-PCR).Minimal inhibitory concentrations(MICs) of ten clinically commonly used antibiotics were determined by agar dilution for AmpC ?-lactamases positive isolates,and the data were analyzed by WHONET 5.3.RESULTS AmpC ?-lactamase phenotype test revealed that 76 AmpC ?-lactamase positive isolates(23.5%) were identified among 324 ESBLs positive isolates of E.coli and K.pneumoniae from four hospitals in Huzhou.69.7% Of the AmpC ?-lactamase phenotype positive isolates were positively amplified by Multi-PCR.The value of MIC50 for carbapenem was lower 0.25 ?g/ml.We also found four carbapenem-resistant strains in this study.CONCLUSIONS We found that the incidence rate of AmpC ?-lactamase is high among the ESBLs producing E.coli and K.pneumoniae strains in Hangzhou.Carbapenem antibiotics have higher antimicrobial activity than other tested antibiotics.

Objective By studying the different blood volume blood donors machine adopt platelet product aggregation,platelet content,the machine adopt circulating volume,etc,analysis of different blood volume machine adopt donors on appropriate conditions.Methods The base material of randomly selected 307 blood donors,through blood related index in the study of circulating volume,blood volume,gender,platelets,Hct,Pct count before collection,machine adopt of platelet aggregation and the relationship between the acquisition time;different blood volume,blood platelet count grouping machine adopt the different of platelet aggregation rate,different amount of platelets collect blood volume group,the comparison of blood circulation.The use of statistical software SPSS 17.0,data analysis,analysis including multiple regression analysis,chi-square test,t test,etc.Results 1)The lower count of platelets,Hct,Pct count before collection,the longer the acquisition time,and gender and Pct has nothing to do with the acquisition time the longer the acquisition time,the higher the machine adopt the possibility of platelet aggregation.2)Low blood volume and low platelet count group,machine mining platelet aggregation rate is higher.3)Machine adopt donors of blood volume is higher,the machine adopt the platelet collection amount,the more and the less blood circulation.Conclusion According to the different blood volume blood donors check-up indicators,further carries on the reasonable analysis,optimized machine mining scheme,especially to reasonable arrangement of low blood volume blood donors,so as to improve the quality of platelet collection.

Objective To study the degradation of hepatitis C virus(HCV) genome before and after methylene blue photochemistry(MB-P) treatment of the plasma.Methods MB was added to HCV positive plasma to a final concentration of 1.0?mol/L.The plasma was then exposed to 30000 Lux fluorescence.Plasma samples were then collected at different times of exposure.Real-time PCR was used to quantitatively study the course of the HCV-RNA degradation.The whole genome was screened for integrity by RT-PCR with 8 pairs of specific primers which targeted sequential overlapping sections of HCV genome.Results Results of real-time PCR showed that the copy number of HCV-RNA was continuously decreasing during MB-P treatment.RT-PCR results showed that the reactivity of different sections of the HCV genome to MB-P was significantly different and indicated that the 5' end and 3' end were more stable. Conclusion MB-P treatment could degrade HCV RNA with various sensitivities to different sections of the genome.RNA degradation may play an important role in plasma virus inactivation.Detection of HCV genomic RNA might be clinically useful to monitor the process and efficiency of virus inactivation.

Objective To study the efficacy of treatment of proximal humeral fractures in elderly patients using cloverleaf plate combined with bone cement. Methods 13 aged patients with serious proximal humeral fractures were admitted from 2000.1 to 2001.6. Their X ray revealed displacement and crispation. After anterior reduction through the shoulder joint, the obvious osseous defects were filled with bone cement. Then the fractures were fixed with cloverleaf plates. Sling was applied for the patients postoperatively and the rehabilitation program started 3 weeks after operation. Results All the 13 cases were followed up. The follow up ranged from 5 to 10 months with an average of 8.3 months. The range of motion was satisfactory with a mean abduction over 90 degrees. X ray revealedthat no patient demonstrated osteonecrosis of the humeral head. Union was achieved in all the patients. According to Neers scale, the function of shouder was excellent in 10 cases, and good in 3 cases. Conclusion This methods is recommended for aged patients with osteoporosis, serious osseous defects and difficulty of using internal fixation alone, because it is convenient and reliable and can prevent replacement of joint prosthesis.