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 investigate the composition and diversity of acaroid mite community in the house in Xuancheng area in Anhui province.Methods The dust samples from 4 sites selected in each house were collected in June-July,2006,acaroid mites were isolated,counted and identified.Results 3 386 acaroid mites were obtained,belong to 6 families,12 genera and 14 species.The species richness index Rmargalef for the four habitats ranged from 0.73 to 1.75,while the species diversity index(Shannon-Wiener index) ranged from 1.36 to 1.87 and Pielou index ranged from 0.70 to 0.92.Conclusion The results of the present paper show that the habitat condition is the main factor that can directly influence the community structure and diversity of acaroid mites in the houses and the human activity is also an influencing factor to be reckoned with.

Objective To observe the efficacy and safety of intravitreal injection of conbercept for wet age-related macular degeneration.Methods Retrospective clinical study.Sixty eyes of 60 cases diagnosed as wet age-related macular degeneration (AMD) definitely were intravitreally injected conbercept 0.5 mg from March 2014 to March 2016.Follow-up time was 6 months to 12 months,averaged (8.5 ± 2.4) months.Visual acuity (ETDRS charts letter),central macular thickness (CMT),leakage of choroidal neovasculaxization (CNV) and area of CNV and complications before and after the treatment were analyzed.Results Conbercept injection therapeutic times were 1-5,the average therapeutic times were 2.80 ± 0.89.At 1 month,2 months,3 months and 6 months after treatment and the last follow up,the mean letter of ETDRS charts increased (14.76 ± 5.89) letters,(19.88 ± 7.13) letters,(24.75 ± 6.74) letters,(23.94 ±6.15) letters,(22.89 ± 8.53) letters compared with pre-operation,respectively (all P ＜0.05).CMT decreased (70.19 ± 60.56) μm,(82.07 ± 57.97) μm,(95.40 ± 87.92) μm,(97.57 ± 46.68) μm,(107.46 ± 56.82) μm compared with pre-operation,respectively (all P ＜ 0.05).Before treatment,the area of CNV leakage was (12.32 ± 5.67) mm2,and at postoperative 3 months,6months the area were statistically significant different (all P ＜ 0.05).Four cases had subconjunctival flake bleeding,intraocular pressure increased slightly in 3 cases and they all recovered at 1 week after treatment.There was no serious ocular and system complication.Conclusion Intravitreal injection of conbercept for wet AMD can improve the visual acuity,decrease CMT and inhibit the CNV regression without serious complication.

OBJECTIVE:To optimize the extraction technology of polysaccharides from Cipangopaludina chinesis by enzyme method. METHODS:The papain enzymolysis extraction was performed. On the basis of single factors tests,taking the yield of polysaccharides as the index,four factors including enzyme concentration,pH,enzymolysis temperature and enzymolysis time in the extraction process were optimized by the orthogonal test,and verification was carried out. RESULTS:The optimal technology by enzyme method was as follows as enzyme concentration of 1.0%,pH of 7.0,enzymolysis temperature of 55 ℃ and enzymoly-sis time of 3.0 h. Under this technology,the average yield of polysaccharides was 4.98%(RSD=2.51%,n=3)and the content of polysaccharides was 73.54%(RSD=1.36%,n=3). CONCLUSIONS:The optimal extraction technology of polysaccharides from C. chinesis was simple,reasonable and feasible.

AIM:To investigate the mechanism of apoptosis during the process of adult rat mesenchymal stem cells(MSCs)differentiating into neuron-like cells in vitro.METHODS:The MSCs were isolated primarily from adult rats bone marrow by density gradient and then expanded in medium as undifferentiated cells for 3-5 generations.The MSCs were divided into three groups at random.The control group was induced with ?-mercaptoethanol(?-ME).Meanwhile,the U0126 group was given ?-ME and U0126(10 ?mol/L)added at the beginning of induction.The PMA groups were treated with ?-ME and different concentrations of PMA since pre-induction.The effects of U0126 and PMA on the activity of neuron-like cells were observed by MTT.The effects of U0126 and PMA on the expression of neuron specific marker nestin and expression of apoptosis-related protein caspase,Bcl-2,Bax in neuron-like cells were detected by using immunocytochemistry method.TUNEL technique was used to detect apoptosis index.RESULTS:Compared to control group,neuron viability,nestin and Bcl-2 increased and neuron apoptosis decreased in U0126 group(P

OBJECTIVE: To investigate the effect of trichostatin A (TSA),a specific inhibitor of histone acetyltransferase,on apoptosis of human hepotoma cell line HepG2,the expression of fragile histidine triad (FHIT) and apoptosis-related proteins in order to study apoptosis mechanisms.METHODS: Human hepatoma cell lines HepG2 were cultured and treated with different concentrations of TSA(125,250,500,1 000,2 000 nmol?L-1) for 24 and 48 hours.Human hepatoma cell lines HepG2 survival and apoptosis were determined by MTT assay with absorbance vale and inhibitory rate as index.Apoptotic percentage of HepG2 treated with 250 and 1 000 nmol?L-1 TSA were determined using TUNEL assay.The expressions of FHIT and caspase-3,bax,bcl-2 were analyzed by immunocytochemistry with solvent as control.RESULTS: As compared with control group,absorbance vale of human hepatoma cell lines HepG2 were decreased after treated with different concentration of TSA in dose-dependent and time-dependent manners.Positive cell rate in TUNEL was increased (P0.05).CONCLUSION: TSA may inhibit proliferation and promote apoptosis of hepatoma HepG2 cell by up-regulating the protein expression of FHIT,caspase-3 and bax.

Objective To investigate the problems existing in hospital transfusion records,analyze unreasonable factors,and improve the quality of blood transfusion.Methods Investigation of our hospital from January 2016 to January 2017 with blood transfusion medical records,according to hospitalization number,900 cases were selected,using random number table.Investigating blood transfusion records on each record integrity,according toGuidelines for Surgical and Traumatic Blood Transfusion and Internal medicine transfusion guide in clinical blood transfusion technical specifications.The common blood components such as red blood cells,plasma and cryoprecipitate infusion and rationality evaluation.Results There were 583 surgical departments and 557 non-surgical departments.There was a significant difference between the surgical department and the non-surgical department.The reasonable rate of non-operation department was higher than that of the operation department (x2 =7.723,P=0.021).The rational rate of the department was 93.8％,while the operation department was only 88.0％;900 blood transfusion records of four kinds of blood components of the rationality of the difference was statistically significant (x2=214.767,P＜0.001).Non-surgical department of erythrocyte,plasma,cold precipitate reasonable rate than the surgical department;900 medical records in 55 records failed,mainly after the assessment of incomplete blood transfusion,no recorded blood transfusion reaction.There were significant differences in the failure rate between the surgical department and the non-surgical department (x2 =4.613,P=0.032).Conclusion Some physicians transfusion indications,for evaluation before and after blood transfusion blood insufficient attention to curative effect evaluation,blood transfusion is not reasonable,and in the operation department,do not take the blood transfusion medical record writing,hospitals should strengthen the blood transfusion blood transfusion continuously improve the scientific and normative management.

Objective To investigate the effect of zerumbone on the proliferation and apoptosis of human pancreatic cancer cell line PANC1 and its possible mechanism.Methods Zerumbone of various concentrations (3.75, 7.5, 15, 30, 60 μg/ml) were used to treat PANC1, and cells without treatment were used as control.CCK-8 assay was used to detect the inhibitory rate of cell proliferation.Cell apoptosis analysis was determined by using Hoechst 33342 staining and flow cytometry.Western blotting was performed to evaluate the phosphorylation Statl ( p-STAT1 ), and Bax and Bcl2 protein expression.Results Zerumbone caused a time- and dose-dependent reduction of cell viability in PANC1 cells.After 48h treatment of Zerumbone of 15 μg/ml, cells inhibitory rate was increased to (72.8 ± 2.72 )%, and classic apoptosis morphology was observed, with apoptosis rate was 14.2%.At the same time, p-STAT1, and Bax protein expression was significantly increased (0.654 ±0.048 vs 0.074 ±0.011, 0.577 ±0.044 vs 0.218 ±0.027,P＜0.05);Bcl-2 protein expression was significantly decreased (0.162 ± 0.029 vs 0.459 ± 0.034, P＜0.05).Conclusions Zerumbone may inhibit the proliferation of PANC1 cells and inducing cell apoptosis,which may be related to the up-regulation of STAT1's activity and Bcl-2/Bax ratio.

Objective To investigate the protective effect of rosiglitazone on acute necrotizing pancreatitis (ANP) associated lung injury in rats.Methods Seventy-five male Wistar rats were randomly divided into sham operation group (SO group),acute necrotizing pancreatitis group (ANP group) and rosiglitazone pretreatment group (ROSI group).ANP model was induced by retrograde infusion of 5％ sodium taurocholate into the biliopancreatic duct.Thirty minutes after ANP induction,ANP groups were injected with 10％ DMSO (0.2 ml/100 g) through femoral vein,and ROSI group were injected with ROSI dissolved with 10％ DMSO (6 mg/kg) through femoral vein,while SO group was injected with normal saline,and 30 minutes later was injected with same amount of 10％ DMSO.Rats were sacrificed at 3 h,6 h and 12 h after the operation.Serum amylase and lung wet/dry weight ratio (W/D) were measured,lung tissues were harvested for pathologic examinations.STAT1 protein and phosphorylation-STAT1 protein (p-STAT1) expression were detected by Western blot.Results The serum levels of amylase,lung W/D,pathologic score of lung tissues in ANP group were increased with time,and reached the peak at 12 h,which were (5017 ± 203)U/L,3.12 ±1.30,(3.33 ±0.18) score,and these were significantly higher than those in SO group (P ＜ 0.05 or 0.01),the expression of STAT1 protein was not statistically significant,but the expression of p-STAT1 reached the peak at 3 h (5.23 ± 0.03),then it gradually decreased,but it was still significantly higher than that in SO group (0.16 ± 0.04,p ＜ 0.01).The serum levels of amylase,lung W/D,pathologic score of lung tissues in ROSI group at 12 h were (1912 ± 164) U/L,1.83 ± 1.26,(2.78 ± 0.16),which were significantly lower than those in ANP group (P ＜ 0.05).The expression of STAT1 protein was not statistically significant,and the expressions of p-STAT1 at 3 h,6 h,12 h was 0.41 ±0.04,0.22±0.05,0.15 ±0.03,which were significantly lower than those in ANP group (P ＜ 0.05).Conclusions Rosiglitazone has the protective effect on ANP associated lung injury by inhibition of phosphorylation-STAT1 protein expression in the early phase.

Objective To evaluate the effects of propofol anesthesia on hippocampal protein kinase A (PKA)-cAMP response element binding protein (CREB) signaling pathway in neonatal rats.Methods One hundred and seventy-five male Sprague-Dawley rats,aged 7 days,weighing 8-15 g,were randomly divided into 5 groups (n =35 each) using a random number table:control group (C group) and propofol 25,50,100 and 200 mg/kg groups (P~ groups).Groups P1 and P2 received intraperitoneal propofol 25 and 50 mg/kg,respectively.Groups P3 and P4 received intraperitoneal propofol 100 and 200 mg/kg,respectively,and after righting reflex completely recovered,an increment of propofol 50 mg/kg was given until the total amount was finished.Five animals in each group were chosen and arterial blood samples were obtained immediately after the animals were fully awake for blood gas analysis.Five animals in each group were chosen at 2 h after fully awake and the age of 9 weeks,the rats were sacrificed and their brains were removed for microscopic examination of the ultrastructure of hippocampal neurons and for determination of PKA mRNA,CREB mRNA,PKA protein and pCREB protein in hippocampus (using RT-PCR and Western blot analysis).Results There was no significant difference in the indexes of blood gas analysis anong the five groups (P ＞ 0.05).Nuclear swelling and fragmentation,chromatin condensation,apoptotic bodies,decreased number of synapses and widened synaptic space were observed in P2,P3 and P4 groups.Compared with group C,the expression of PKA mRNA,CREB mRNA,PKA protein and pCREB protein was significantly down-regulated at 2 h after fully awake and the age of 9 weeks in P1,P2,P3 and P4 groups (P ＜ 0.05).Conclusion The mechanism by which propofol anesthesia induces neurotoxicity in neonatal rats may be related to inhibition of the activity of PKA-CREB signaling pathway.