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 pathogenic molecular mechanism of H.pylori through analyzing its effect of on MMP-9 expression. Methods BGC-823 cells were cocultured with ATCC49503(CagA +)and HP 030811 (CagA -) H.pylori strains respectively, then the total cellelar RNA was extracted. The MMP-9 mRNA expression in BGC-823 cells, 9 cases of H.pylori-positive and 9 cases of H.pylori-negative gastric ulcer biopsy tissues was detected by RT-PCR. The MMP-9 protein expression on the paraffin-imbedded tissue sections of 17 cases of H.pylori-positive and 14 cases of H.pylori-negative gastric ulcer tissues was detected with immunohistochemical technique, and was quantified by image quantitative analysis. Results H.pylori induced MMP-9 expression in BGC-823, and the level of MMP-9 expression induced by CagA -postive H.pylori strain was higher than that by CagA -negative one. MMP-9 expression level was higher in the H.pylori-positive gastric ulcer tissues than that in the H.pylori-negative ones. Conclusion H.pylori-induced MMP-9 expression may play an important role in gastric ulcer formation and gastric carcinogenesis.

Objective To Study the method for determination of aluminium in water by spectrophotometry of Be ryllon Ⅲ. Methods Be ryllon Ⅲ reacts on aluminium in oppropriate temperature and acidity in water and produces a red complex compound?The red complex compound can be determined by spectrophotometry. Results The optimum reaction temperature was 85 ℃, the optimum reaction time was 5 min, pH of buffer was 6.1, Beer's law was obeyed in the concentration range of 0-0.02 mg/L for aluminium, rates of recovery were 80.0%-96.0% and RSD was 0.70%. Conclusion This method is accurate, simple, sensitive and repeatable.

Objective:To explore the early diagnostic value of joint detection heart type fatty acid binding protein (h-FABP), troponin (cTnI), myoglobin (Mb) and creatine kinase isoenzyme Mb (CK-Mb) in children's hand, foot and mouth disease (HFMD) combined myocardial injury.Methods: Choice 276 cases of HFMD as observation group, and 40 healthy children as control group. Were determined h-FABP, cTnI, Mb and CK-Mb content in serum at different time, analysis of various index level differences and dynamic change between groups in different period.Results: Among 276 patients with HFMD, 57 cases of diagnosed myocarditis, concurrent rate was 20.65%. Within 0~3 hrs, abnormal rate of h-FABP, cTnI, Mb and CK-Mb in serum were 20.29%,1.81%,14.86% and 2.90%, in 276 cases of children with HFMD. The abnormal rate of h-FABP and Mb was obviously higher than that of cTnI and CK-Mb, the results between the difference was statistically significant (x2=35.132,x2=37.063,P<0.01),h-FABP abnormal rate is higher than CK-Mb, the difference was statistically significant(x2=3.175,P<0.05). the,serum h-FABP cTnI, Mb and CK-Mb concentrations in children of HFDM combined with suspicious viral myocarditis were significantly higher than that of control group, the difference had statistical significance (t=37.625,t=23.172,t=17.261,t=18.724,P<0.01). H-FABP and Mb concentration on HFMD combined myocarditis began to rise after the occurrence of 0~3 h, 4~9 h to peak, CTnI and CK-MB 4~9 h to rise, 10~12 h to peak, has been in a higher level in the 12~72 h.Conclusion: HFMD combined myocarditis had a higher incidence, h-FABP is the most sensitive indicator of early diagnosis, followed by Mb. CTnI and CK-MB are parameters of diagnosis sensitivity for HFMD combined with myocarditis in middle-late period.

Objective:To evaluate the clinical effect of the revaseularization procedure for immature teeth with necrotic pulp and apical periodontitis.Methods:12 immature teeth with necrotic pulp and apical periodontitis in 12 patients(9-13 years old) were treated with revascularization protocol.The patients were followed up for 21-24 months.Results:All teeth were cured.4 types of responses were observed:①Increased thickening of the canal walls and continued root maturation;②Severe calcification of the canal space beneath MTA;③Partial pulp canal obliteration;④Continued root development with an open apex.Conclusion:Revascularization procedure may the formation of the root apex but not in all the cases.Excessive deposition of hard tissue might caused calcification in canal space.

Objective To observe the analgesia effect of combination of continuous femoral nerve block and oral analgesics after total knee arthroplasty and the impact on early rehabilitation of knee.Methods Sixty patients scheduled for unilateral total knee arthroplasty were randomly divided into two groups.One group was the multimodal analgesia group (group M)and the other was patient controlled intravenous analgesia group (PCIA)(group Ⅰ),30 patients in each group.In group M, patients were given celecoxib (200 mg,bid)and oxycodone (10 mg,bid)two days before operation, then received continuous femoral nerve block followed by oral acetaminophen (100 mg,tid)and oxyc-odone (20 mg,bid)three days after operation.In group I,patients took no medicine before operation and only received PCIA after operation.The two groups were compared with postoperative visual ana-logue scale(VAS)scores at 6,12,24,48 h at rest,during active training and continuous passive movement,range of motion of the knee joint during active training and continuous passive movement, the first time of ambulation,length of hospital stay,range of motion of the knee joint at hospital dis-charge and the incidence of complications.Intravenous dezocine 5 mg was given when VAS score was higher than 5,which was recorded.Results Compared with group Ⅰ,the VAS scores at rest,during active and continuous passive movement at postoperative 6,12,24,48 and 72 h in group M were de-creased significantly (P <0.05).The range of motion of the knee joint during active training and con-tinuous passive movement at postoperative 24,48 and 72 h in group M was increased significantly(P<0.05).The first time of ambulation(2.5±0.8)d and length of hospital stay(9.1±2.3)d in group M were shorter significantly than the first time of ambulation (3.3 ±0.7)d and length of hospital stay (10.8±2.0)d in group I (P <0.05 ).The range of motion of the knee joint(95.6 ± 1 7.2)°in group M at hospital discharge was greater significantly than the range of motion of the knee joint (82.5± 1 5.2 )° in group I (P < 0.05 ).The incidence of nausea and vomiting (3.3%),uroschesis (6.7%)in group M was lower significantly than the incidence of nausea and vomiting (30.0%), uroschesis (26.7%)(P < 0.05 ).Additional use dezocine (3.3%)was lower significantly than additional use dezocine (40.0%)(P <0.05).Conclusion Compared to patient controlled intravenous analgesia,combination of continuous femoral nerve block and oral analgesics can effectively alleviate postoperative pain in patients undergoing total knee arthroplasty,which is beneficial to early postop-erative knee function recovery.

Objective:To establish a quantitative method for the determination of lysine hydrochloride in pediatric compound lysine granules. Methods:The quantitative determination was performed on an amino acids analyzer. The cationic resin column(150 mm × 4. 6 mm,7 μm)was used. The mobile phase was citrate buffer solution (pH 3. 45) and citrate buffer solution (pH 10. 85) with gradient elution at a flow rate of 0. 45 ml·min-1 . The detection wavelengths were set at 570 nm and 440 nm. Results:The linear range of lysine hydrochloride was 2. 692-21. 536 μg·ml-1(r=0. 999 5), and the average sample recovery was 98. 85% with RSD of 0. 63% (n=9). Conclusion:The method is sensitive, simple and reproducible. It can be used for the quality control of pediatric compound lysine gran-ules.

Objective To investigate the distribution and antibiotic resistance patterns of uropathogens in children with complicated urinary tract infection to guide the empirical antimicrobial treatment.Methods Retrospective analysis of isolated bacteria from 141 children hospitalized with complicated urinary tract infection was performed from January 2011 to January 2013.Results Gram-negative bacillus were the main pathogens [56.6％(81/143)],and Escherichia coli was the leading cause (41.3％,59/143).Gram-positive bacillus accounted for 38.5％ (55/143),including Enterococcus faecium (17.5％,25/143) and Enterococcus faecalis (14.7％,21/143).Fungi caused 4.9％ (7/143) of the infections.The resistance of Escherichia coli to amoxicillin was the highest [92.0％ (23/25)] and resistance to imipenem (4.3％,2/47),amikacin (6.8％,4/59) and nitrofurantoin (6.8％,4/59) was lower.It was observed resistance of Enterococcus faecium to linezolid,quinupristin,vancomycin and nitrofurantoin was significantly lower than other common antibiotics.The resistance of Enterococcus faecalis to quinupristin,tetracycline,erythromycin,rifampicin was significantly higher than other common antibiotics.Concluusions Escherichia coli,Enterococcus faecium and Enterococcus faecalis are the common uropathogens.These pathogens have a high antibiotic resistance to many common antibiotics.The selection of empirical treatment of children with complicated urinary tract infection should be adjusted by antimicrobial susceptibility test.

Objective:To analyze the osmotic pressure of citicoline sodium injection from different manufacturers to provide experi-mental basis for improving the national standard of the product. Methods:The cryoscopic method was used to detect the osmotic pres-sure of 479 batches of citicoline sodium injection from 45 pharmaceutical factories. Results:The osmotic pressure of citicoline sodium injection was within the range of 359-692 mOsmol · kg-1 , that of citicoline sodium for injection was within the range of 401-1408 mOsmol·kg-1 , and that of citicoline sodium chloride injection was within the range of 278-299 mOsmol·kg-1 . Conclusion:The os-motic pressure of citicoline sodium injection and citicoline sodium for injection from different manufacturers is obviously different. It is necessary to detect osmotic pressure in the quality standard in order to evaluate the quality of preparation process.

Objective To assess the characteristics of different doses of cisplatin-induced acute kidney injury,further to understand mitochondrial dysfunction and its role in acute kidney injury (AKI).Methods Male C57BL/6J mice were first randomly divided into two groups:control group (n =6) and AKI group (n =12).Then,AKI group was subsequently divided into other two groups according to different dose of cisplatin (10 mg/kg or 20 mg/kg).AKI group received intraperitoneal injection of cisplatin.All mice were sacrificed after 72 h of injection.Renal biochemical function,renal pathological changes,renal injury markers,kidney mitochondrial function and structural changes were observed.Results (1) After 72 hours of injection,the AKI group performed significant kidney injury changes compared to control group,thereinto 20 mg/kg group was more serious than 10 mg/kg group.With the cisplatin dose increasing,renal function markers such as serum creatinine,urine protein gradually increased.(2)Kidney biopsy showed tubular structural damage,the formation of protein casts,kidney injury molecule-1 (KIM-1) gradually increased(P ＜ 0.05).(3)Electron microscopy found tubular mitochondrial structural damage,mtDNA copy number decreased,the level of peroxisome proliferatoractivated receptor-gamma coactivator-1alpha (PGC-1α),ATP synthase β decreased(P ＜ 0.05),and Western blotting manifested cytochrome C was released from mitochondria to the cytoplasm.These data all exhibited significant difference between different groups(P ＜ 0.05).Conclusions Cisplatin induces acute kidney injury in dose-dependent manner.Mitochondrial dysfunction participates in kidney injury,and is also related to the kidney pathological damage.