Microfluidics-based digital PCR depends on microfluidic chip to split PCR reaction mixture into many tiny equal-volume units.Quantitative assessment of target DNA template can be obtained by counting the number of fluorescence-positive units after thermocycling.Microfluidics-based digital PCR exhibits many advantages including absolute quantification, high sensitivity and accuracy, and shows great promise in a variety of applications, such as infectious diseases diagnose, early cancer detection and prenatal diagnose. There are already several microfludics-based digital PCR products produced from sereval companies.It is believed that as the technology improves, microfluidics-based digital PCR will find broader applications and become the next-generation tool for genetic tests.

Objective To study the impact of HBV/C gene G87 mutation on HLA-I expression of host cells. Methods HBV wild-type genome plasmid was reconstructed by site-directed mutagenesis technique and subcloning technique into expression vector of G87 mutant (EBO-G87) and expression vector of wild type (EBO-WT), which were transfected into HepG2 cells via liposome technique,respectively.The transfected cells were stained with murine mAb anti-HLA-ABC conjugated directly to FITC, and HLA-I expression on their membranes was analyzed by flow cytometry. Results The constructed vectors of EBO-WT and EBO-G87 were identified by restriction enzyme digestion and nucleotide sequencing. The mean fluorescence intensity of HLA-I expression on transfected cells with EBO empty vector was at low level(2.3). It was elevated remarkably to 18.8 by EBO-WT, while that of EBO-G87 was increased to 10.5. Conclusion Two strains of HBV may up-regulate the expression of HLA-I, and G87 mutation of C gene may affect the expression level of HLA-I on host cells.

Objective To evaluate the changes parameters of polysomnographic ( PSG) with different subtypes of primary Parkinson's disease ( PD ) and analyze the clinical characteristics of sleep disorders in PD patients. Methods Ninety patients with primary PD [ tremor-predominant PD group ( n = 40 ) , postural instability gait disorder ( PIGD )-predominant PD group ( n = 50 ) ] and 50 healthy controls were detected by full night PSG monitoring. And the results were compared. Results Compared with tremor-predominant PD group, the total sleep time, bed time, sleep wake-up times, sleep efficiency and rapid eye movement ( REM ) sleep time of PIGD-predominant PD group were significantly lower (all P<0. 05), however, the sleep latency, S1 (min), S1 (%), S2 (min), S2 (%), S3+S4(min), S3+S4 (%) had no statistical significance (all P>0. 05). Compared with the controls, the total sleep time and bed time of tremor-predominant PD group were significantly lower (all P<0. 05), however, the sleep wake-up times, sleep efficiency, REM sleep time, sleep latency, S1 ( min ) , S1 (%) , S2 (min), S2 (%), S3 + S4 (min), S3 + S4 (%) had no statistical significance (all P>0. 05). Combined with video surveillance video observation,the abnormality of REM sleep behavior disorder ( RBD) was 70% with PIGD-predominant PD group, in which the abnormality of tremor-predominant PD group was 2. 5%. Conclusions The PIGD-predominant PD patients are more likely to appear sleep disorders and RBD than tremor-predominant PD patients. It may be related to clinical heterogeneity of different PD subtypes.

OBJECTIVE To investigate the pathogenic causes of community acquired pneumonia(CAP) in the elderly and to guide experience antibiotic therapy.METHODS An etiological study was performed on 204 elder patients with CAP in the Cadres Medical Department of Beijing Shijitan Hospital from May 2005 to Apr 2008.The count scores were by CURB-65 scoring system.RESULTS Pathogens were identified in 204 patients: there were Mycoplasma pneumoniae(42,20.6%),Streptococcus pneumoniae(30,14.7%),Haemophilus influenzae(24,11.8%),Chlamydia pneumoniae(17,8.3%),Klebsiella pneumoniae(14,6.9%),Escherichia coli(7,3.4%),Pseudomonas aeruginosa(5,2.5%),Staphylococcus aureus(4,2.0%),Legionella pneumophila(2,1.0%),and Acinetobacter baumannii(1,0.5%).The score,by CURB-65 scoring system,of mixed infected patients was higher than single infections.CONCLUSIONS Atypical pathogens have important role in elderly CAP,the most common pathogenare Gram-negative bacilli,S.aureusand fungi.Mixed infection couldn′t be ignored.

Objective:To investigate the expression of IGF-IR and PKC and their clinical significance in laryngeal squamous cell carcinoma(LSCC).Method:The expression of IGF-IR and PKC protein were detected by immunohistochemistry and Western blot in 60 cases of LSCC, 25 cases of adjacent non-tumorous laryngeal epithelium and 10 cases of normal laryngeal epithelium.Result:The positive rate of IGF-IR and PKC expression in LSCC were significantly higher than that in normal laryngeal epithelium and adjacent non-tumorous laryngeal epithelium(P<0.05). IGF-IR positive expression were increased more in cases with stage Ⅲ-Ⅳ, cervical lymph node metastasis and well differentiation than that in cases with stageⅠ-Ⅱ, non-lymph node metastasis and poor/mediate differentiation, respectively(P<0.05).The positive rate of PKC in LSCC were correlated with clinical stage and cervical lymph node metastasis.There was a correlation between the expression of IGF-IR and PKC in LSCC.Conclusion:The overexpression of IGF-IR and PKC in the laryngeal carcinoma may play an important role in the carcinogenesis and development of LSCC. It is suggested that detecting the expression of IGF-IR and PKC can be used for early diagnosis and treatment of LSCC.

Objective To observe the effect of S-band micro-wave long-term intermittent irradiation on endocrine function in rats.Methods A total of 192 rats (male and female) were randomly divided into the sham-irradiation (normal control) groups and the irradiation groups.The irradiation groups were exposed with micro-wave at 3 dosages of 4,10 and 20 mW/cm2 for 6 min twice a week for 12 weeks,while no administration was given to control group.The endocrine parameters in blood serum were examined by radioimmunoassay at 4,8,12 week during irradiation and 4 week post-irradiation.Results After the irradiation of S-band microwave,parts of the endocrine parameters changed.T3 in famale rats decreased at first and then increased,especially in 10 mW/cm2 group at 8 and 12 week,20 mW/cm2 group at 4 and 12week(t =-2.586,-2.642,-5.075,-4.365,P <0.05).FT3 in famale rats had the similar trend asT3,significantly lower in 4 and 10 mW/cm2 groups than that in the control group at 4 week (t = 2.275,2.510,P <0.05),then increased,especially in three irradiation groups at 12 week (t =-2.636,-2.851,-5.240,P < 0.05).TSH decreased at 4 week,especially in 10 mW/cm2 group (t = 2.300,P < 0.05) ; and then increased in the irradiation groups at 20 mW/cm2 at 8 and 12 week (t =-2.838,-3.651,P <0.05).COR and ACTH in male rats showed changes in volatility,in which the 4,10 and 20 mW/cm2 groups at 8 week increased significantly (t =-2.772,-2.234,-2.505,P < 0.05),while 20 mW/cm2 group at 12 week decreased significantly (t=3.067,P < 0.05).E2 in female rats was slightly lower in irradiation groups at 4 week than the control group,then increased,especially in 10 mW/cm2 group at 8 week,three irradiation groups at 12 week (t =-2.322,-3.179,-2.655,-4.716,P < 0.05),and returned to the normal at 4 week post-irradiation,significantly lower in 4 mW/cm2 group than that in the control group (t = 2.250,P < 0.05).T in male rats increased first and then decreased,especially in 10 mW/cm2 group at 8 week(t =-2.435,P < 0.05).After exposure the above indexes restored to some extent.Conclusions The long-term intermittent irradiation of S-band microwave can cause adverse effects on the endocrine function of rats.

Objective To evaluate the value of plasma soluble urokinase plasminogen activator receptor (suPAR) and serum pmcalcitonin (PCT) to investigate their assessment of disease severity and prognosis in patients with sepsis.Methods The levels of plasma suPAR and serum PCT were monitored in 77 patients with sepsis.The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) score were recorded.According to the disease severity and their prognosis,the value of plasma suPAR,serum PCT,APACHE Ⅱ and SOFA score on predict the disease severity and prognosis of septic patients were compared.Results The levels of plasma suPAR in septic patients [(7.9 ±6.5) ng/mL] were lower than severe sepsis patients [(8.4 ±4.5) ng/mL] and septic shock patients [(13.9 ± 8.0) ng/mL],allP ＜ 0.05.The levels of serum PCT in septic patients (6.3 ± 3.5) ng/mLwere lower than severe sepsis patients [(23.7 ± 3.9) ng/mL] and septic shock patients [(25.7 ±4.3) ng/mL],allP ＜0.05.But there was no significant difference in the levels of serum PCT between the severe sepsis group and the septic shock group.Receiver operator characteristic curve (ROC)of the level of plasma suPAR could distinguish survivors from non-survivors in septic patients,maximal area under curve (AUC) of plasma suPAR was 0.803.The best cut-off value of plasma suPAR to distinguish survivors from non-survivors was 9.905 ng/mL.And the AUC of serum PCT was 0.61 (P ＞ 0.05) ; the AUCofAPACHEⅡ score was 0.832 (P＜0.05); the AUC of SOFA score was 0.767 (P＜0.05).Conclusion Monitoring of the levels of plasma suPAR and the APACHE Ⅱ score can help to assess the severity and the prognosis of sepsis in the early stage.

This paper discussed that how to cultivate high-quality medical talents by the use of basic education,medical moral education and psychologic education etc.

Objective To develop a connector between anesthetic machine and oxygen provider for continual oxygen supply to anesthetic machine during field operation.Method According to the principle of three-way block and one-way qas wave,the new connector with two-routes but one-way for the anesthetic machine was made.Result This setting enable the anesthetic machine to connect two oxygen providers at one time.Conclusion With low cost and being easy to install and dissemble,it can be connected to all kinds of anesthetic machine,thus ensuring the persistence work of the anesthetic machine when changing oxygen provider.

OBJECTIVE:To provide reference for improving the application ability of professional English for students major-ing in pharmacy in higher vocational education. METHODS:The career-oriented practical pharmaceutical English course and teach-ing system were constructed through the way of formulation of curriculum standards,establishment of teachers’team,development of school-based teaching material,application of CBI theme teaching mode and variety teaching methods,and it was carried out among students majoring in pharmacy in 2011-2014 grades in our school. Besides,experimental and survey research were used for the evaluation of Proctical Pharmaceutical English. RESULTS:There was no significant difference in the academic achievement and questionnaire evaluation between 2 class before teaching(P>0.05);however,academic achievement and questionnaire evaluation were higher than 2 class after teaching,experimemal class higher than control class,the difference was statistically significant(P<0.05). CONCLUSIONS:The career-oriented Practical Pharmaceutical English course and teaching system is feasible and effective;compared with traditional teaching,it can inspire students' interest in English learning,improve the ability to use English in the work and motivate their comprehensive qualities.