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 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.

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.

ObjectiveTo determine the levels of 8-iso-prostaglandin F2α (PGF2α) in sera and lesions as well as antioxidant capacity in sera of psoriatic patients,and to assess their correlations with disease severity.MethodsSerum and skin tissue samples were collected from 15 healthy controlsand 50 patients with psoriasis vulgaris.Spectrophotometry was performed to determine the levels of total antioxidant capacity (T-AOC) and the activities of antioxidant enzymes such as superoxide dismutase (SOD),catalase (CAT) and glutathione peroxidase (GSH-Px) in serum samples.Enzyme-linked immunosorbent assay (ELISA) and immunohistochemical SP method were carried out to detect the expression level of 8-iso-PGF2α in the serum and tissue specimens respectively.ResultsThe psoriatic patients showed a significant decrease in the serum level of TAOC((12.78 ± 7.75) U/ml vs. (23.17 ± 8.81) U/ml,P＜ 0.01) as well as the activities of SOD((28.91 ±9.35) U/ml vs.(51.36 ± 7.92) U/ml,P＜ 0.01) and GSH-Px ((180.64 ± 47.70) U vs.(244.20 ± 66.68) U,P ＜ 0.01 ) compared with the healthy controls.The serum T-AOC level and SOD activity were lower in patients with severe psoriasis than those with mild or moderate psoriasis ((9.06 ± 5.30) U/ml vs. (15.27 ± 8.18) U/ml,(21.63 ± 5.28) U/ml vs. (33.76 ± 8.28) U/ml,both P＜ 0.01 ),while there was no significant difference in the activity of GSH-Px between patients with severe and mild or moderate psoriasis.The serum CAT activity was significantly higher in patients with mild or moderate psoriasis than in the healthy controls and patients with severe psoriasis ( (36.92 ± 11.31 ) U/ml vs.( 28.55 ± 8.51 ) U/ml and (24.15 ± 9.36 ) U/ml,P ＜ 0.05 and 0.01 ).Increased serum and lesional 8-iso-PGF2α levels were observed in psoriatic patients compared with the healthy controls ( (88.77 ± 25.27) ng/L vs.(38.34 ± 8.94) ng/L,0.0186 ± 0.0082 vs.0.0027 ± 0.0014,both P ＜ 0.01),as well as in patients with severe psoriasis compared with those with mild or moderate psoriasis(( 114.24 ±13.93) ng/L vs.(71.78 ± 14.35) ng/L,0.0279 ± 0.0027 vs.0.0125 ± 0.0030,both P＜ 0.01 ).The psoriasis area and severity index(PASI) score was negatively correlated with T-AOC level,SOD and CAT activities(r =-0.384,-0.573 and -0.444,all P ＜ 0.01 ),positively correlated with serum and lesional 8-iso-PGF2α levels (r =0.710,0.783,both P ＜ 0.01 ),and uncorrelated with GSH-Px activity.None of the parameters was correlated with the course of disease.ConclusionThe serum and lesional levels of 8-iso-PGF2α may be a more sensitive marker for oxidative damage and disease severity.

Objective To investigate the carboprost ammonia butyl alcohol three plus motherwort injection on prevention of high risk maternal postpartum hemorrhage effect .Methods 120 cases of cesarean section operation and potential bleeding symptom of women were selected as the research object ,and randomly divided into two groups , the observation group of 60 cases using carboprost ammonia butyl alcohol three joint of leonurus heterophyllus injec -tion for prevention and treatment ,the control group of 60 cases used oxytocin for prevention and treatment ,prevention and treatment effects were compared between the two groups .Results After treatment,the observation group 2h,24h postpartum hemorrhage were (422.41 ±213.49)mL,(35.29 ±16.44)mL,were significantly less than those in the control group (589.64 ±345.21)mL,(69.31 ±29.47)mL(t=6.732,8.915,all P<0.05);the observation group 1 cases without production after bleeding ,control group 5 cases of postpartum hemorrhage ,statistically significant differ-ences between the two groups (χ2 =4.973,P<0.05).Conclusion Carboprost ammonia butyl alcohol three plus motherwort injection on prevention of high risk maternal postpartum hemorrhage has a significant effect ,which can re-duce the cesarean section uterine bleeding and reduce the amount of bleeding ,promote uterine involution ,conducive to maternal rehabilitation .

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.

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.