Aim To establish the standards for evaluation of microcirculation in a rat hind limb ischemia model.Methods Rats were subjected to unilateral femoral artery ligation.After 4 weeks,the regional blood flow in skeletal muscles was measured by fluorescent microsphere assay.Formation of collateral vessels was determined by microangiography.Capillary density was examined by immunostaining for CD34.Results In ischemic limbs,regional blood flow decreased significantly,some collateral vessels formed with typical "corkscrew" appearance,and histological analysis showed lower capillary density.Conclusion Quantitative assessment of microcirculation in a hind limb ischemia model is established.

Objective To investigate the inhibition mechanism of rohdea roth on hyphae formation by Candida albicans.Methods MTT assay was used to detect the minimum inhibitory concentration(MIC)and minimum fungicidal concentration(MFC)of C.albicans.The inhibitory effect of fungicidal adherence was detected by MTT assay.Inverted fluorescence microscope was used to observe effect on hyphae formation.The influence on Efg1 and Hwp1gene expression were detected by RT-PCR method.ResultsMIC and MFC of C.albicans were 16 mg/mL and 32 mg/mL, respectively.The inhibitory effects of rohdea roth on C.albicans adherence and hyphae formation were significantly inhibited,and the concentration was dose-dependent.After the concentration of 16 mg/mL acted on C.albicans for 6 h, hyphae disappeared completely.The results of RT-PCR showed that the gene expression of Efg1 and Hwp1 could be inhibited by rohdea roth.Compared with the control group,the expression of Efg1 and Hwp1in the experimental groupwere reduced by 84.18% and 59.57%(P<0.01).Conclusion The inhibitory effect of rohdea roth on the adherence and hyphae formation of C.albicans is mainly through inhibiting the expression of Efg1 and Hwp1genes.

Objective To explore effect of immune function of children with EB virus infectious mononu-cleosis (IM). Methods From Sep. 2011 to Jun. 2013, a total of 62 cases of children with IM were analyzed retrospectively. The proportion of T-lymphocyte subsets ,B-lymphocytesand and NK cells in EBV-DNA group with positive and negative were compared. Results The numbers of IM EBV-DNA-negative cases were 18 (29.03%), the numbers of IM EBV-DNA-positive cases were 44(70.97%), the averages of EBV-DNA in positive group is 6.1 × 103/mL. CD3+,CD8+cell ratios in IM children are significantly higher than control group (P<0.01), CD4+, CD19+cell ratios are significantly lower than control group (P<0.01), NK cells are not significantly different (P>0.05). In the IM children, CD4+, CD19+cells in DNA-positive group are significantly less than DNA-negative group (P<0.01), CD8+cells are higher than DNA-negative group (P<0.05), NK cells are not significantly different (P>0.05). Conclusions Positive blood EBV-DNA is closely related to changes in T-lymphocyte and B-lymphocyte, but isn′t obviously related in NK cells. So effect of EB virus in IM child have certain clinical values to judge the severity and prognosis.

ObjectiveTo design a new operation approach:extreme lateral transforaminal lumbar interbody fusion (E-TLIF) via digital technology and to discuss its feasibility in treatment of degenerative lumbar diseases.MethodsCT scan image data of lumbar vertebra were obtained from volunteer (a healthy male of 26 years old,67 kg in weight and 172 cm in height).Mimics was used to read and reconstruct the data into 3D images.We observe the anatomical bone structures of intervertebral foramen and nerve roots through CT scan and 3D reconstruction in Mimics to analysis the feature of this area.Then E-TLIF operation,pedicle screw insertion and cage placement were simulated by computer via digital technology in Mimics.The feasibility of this new approach was proved through operating on lumbar of formalin doused body with E-TLIF approach.ResultsAn incision of 9 cm lateral form the central line and 45° from the sagittal plane is operated to fully expose the foraminal area.Remove superior facet while save inferior facet is feasible with this approach in E-TLIF.There is enough room for fully revealing and excision of intervertebral disc,and cage placement through enlarged foraminal area.Digital technology is a more economic,efficient way to design a new operation approach.The results are in accord with the formalin doused body test.ConclusionMimics is an effectively,reliable and economic method to design an operation approach.And the result suggested that E-TLIF is a safe,less minimal invasive and more efficient operation approach in treatment of degenerative lumbar diseases.

Objective To establish a rapid,sensitive and specific assay based on real-time PCR combined with reverse transcription for detecting and identifying viable Listeria monocytogenes.Methods The hlyA gene of Listeria monocytogenes was chosen as target,and then the primers and TaqMan probe were designed.Both ends of probe were modified with two different fluorescence groups.The PCR reaction was optimized systematically.The mRNA of Listeria monocytogenes was extracted,and then reverse transcription was performed through random primer.The cDNA Was detected by real-time PCR.Then the specificity,sensitivity and reproducibility of real-time PCR were estimated.In final,real-time PCR was applied to detect 20 mocked double-blind samplea.Results Viable Listeria monocytogenes were detected by real-time PCR accurately and quickly,and meanwhile,none of other bacteria and non-viable Listeria monocytogenes could be identified.The sensitivity was 10 CFU/ml in pure culture and 103CFU/ml for mocked samples respectively.The coefficient of variation of intra-assay and inter-assay Was less than 5%.When this assay was applied directly to identify 20 mocked double-blind samples,10 of these were positive to viable Listeria monocytogenes,5 were negative to non-viable Listeria monocytogenes,and 5 were negative to other pathogens.Conclusion It is demonstrated that real-time PCR is a reliable,accurate and feasible assay for viable Listeria monocytogenes.The establishment of this assay provided complete data for analysis and diagnosis in the field of food safety and epidemiologic survey.

BACKGROUND:Tantalum rod implant technology is a new method of early osteonecrosis treatment. Current research on stress distribution before and after tantalum rod implant in different sizes of femoral head necrosis area is few. OBJECTIVE:To analyze the stress distribution before and after tantalum rod implantation in different sizes of necrotic femoral head area using three-dimensional finite element method. METHODS:Three-dimensional finite element models of normal femoral head and necrotic femoral head of 15, 20 and 30 mm diameterwere constructed. Eight measuring points were chosen on two tiers of each necrotic model to detect the stress distribution and its alteration before and after tantalum rod implantation. RESULTS AND CONCLUSION:(1) Stress concentration werefound on every necrotic femoral head, most pronounced on the one with 30 mm lesion. (2) Tantalum implant appeared to reduce the stress concentration generaly. Comparison of the peak points of these models indicated most significant benefit in 15 mm lesion, next in 30 mm lesion, last in 20 mm lesion. (3) Results indicate that larger lesion entails more concentrated stress distribution and more likely to colapse. Tantalum rod implantation can delay the development of necrosis of the femoral head, andismost effective in smal lesion.

Objective To study the risk factors of prolonged postoperative recovery after the total cavopulmonary connection(TCPC) in the current era.Methods Data on all patients admitted to the cardiac intensive care unit (CICU) after a TCPC between January 2013 and March 2014 were retrospectively analyzed.We excluded all patients who died and required TCPC takedown.The study cohort was further divided into a prolonged recovery group that included patients with 75％ ile for duration of mechanical ventilation or pleural drainage,and a standard recovery group which included all other patients.A multivariable logistic regression model was used to compare demographic,anatomic,and physiological variables between the prolonged and standard recovery groups.Then,the cohort was separated into a high volume resuscitation group and a low volume resuscitation based on the 75％ ile for volume resuscitation(ml/kg) administered on the first three days after the TCPC.Results Totally 118 TCPC operations were performed.Of the study population (n =118),the median age was 3.8 years (3.1 to 4.8 years) and median weight was 14.8 kg(13.3 to 17.1 kg).The most common diagnosis was double outlet of right ventricle (n =47,39.8％).The extracardiac conduit fenestrated TCPC was the most common surgery(n =79,66.9％).Within the study population,43 (39.8％) patients met criteria for prolonged recovery.Univariate risk factors for prolonged recovery included higher preoperative mPAP(P =0.022),atrioventricular valve regurgitation (P =0.000),longer total bypass time (P =0.044),higher postoperative central venous pressure (P =0.000),AST (P =0.001),ALT (P =0.010),NT-proBNP (P =0.000),SaO2 (P =0.012),I n-otropic score (P =0.001),higher incidence of arrhythmia (P =0.000),low cardiac output syndrome (P =0.000),need for peritoneal dialysis (P =0.000),and requirement for greater volume resuscitation during the 72 postoperative hours(75％ for the entire group,P =0.000).In a multivariable Logistic model,need for greater volume resuscitation (OR 10.860,95 ％ CI 2.681,43.987) and the higher postoperative central venous pressure (OR 1.446,95 ％ C I 1.113,1.879) were the only two independent risk factors for prolonged outcome after the TCPC.Conclusion The need for high volume expansion and higher central venous pressure were the risk factors of mediate prolonged recovery.

Objective:To investigate the relationship between the distance from the upper and lower resection margin and the gastric cancer patients with R0 resection and no distant metastasis.Methods:Retrospective analysis of 281 patients with gastric cancer in our hospital,the relationship between the age,sex,tumor size,tumor size,vascular invasion,lymph node metastasis,TNM stage,type of gastric cancer,tumor location and operation mode was analyzed.To compare the survival time of patients with different upper and lower margins.Results:The increase of the distance from upper resection margin was significantly related to the tumor size＞5 cm,TNM stage,type of gastric cancer,tumor location,and the difference was statistically significant(P＜0.05);The distance of lower resection margin was significantly correlated with tumor size＞5cm,vascular invasion,lymph node metastasis and TNM stage,type of gastric cancer,and the difference was statistically significant (P＜0.05);Themedian survival timein patientsof upper resection marginr□3cm with 48 months of the 5-year follow-up period was significantly higher than that in patients of the resection margin＞3cmwith 46 months (P＜0.001).Themedian survival timein patientsof lower resection margin□3cm with 45 months of the 5-year follow-up period was significantly higher than that in patients of the resection margin＞3cm with 44 months (P＜0.001).Conclusion:Gastric cancer postoperative upper and lower resection margin was significantly related with tumor size,TNM staging factors,and the median survival time of upper and lower resection margin＞5 cm was significantly lower than that of the resection margin3 cm.

Background and purpose:Hepatocellular carcinoma suppressor gene-1(HCCS1)is a potential hepatocellular carcinoma supressor gene,and it also plays an important role in sorting of some cytoplasmic proteins.Its carcinoma suppressor function may be related to its sorting function.So it is important to identify the minimum region of functional sequence in HCCS1 that related to the transportation.Methods:The expression vectors containing various lengths of HCCS1 gene were constructed and transfected into HeLa cells mediated by liposomes.The localizations of different HCCS1 fragments and the colocalizations of M6PR with various truncated HCCS1 were determined by laser scanning confocal microscopy,respectively.Results:10 different expression vectors containing various lengths of HCCS1 gene were successfully constructed,it had been shown that the polar localization of HCCS1 protein and the co-localizations with M6PR disappeared when HCCS1 gene was truncated to 1 120 bp from the 3'end by laser scanning confocal microscopy.Conclusions:We identified the minimum localization of the HCCS1 functional sequence that related to the transportation.

Background and purpose:Wuxing soup is popular for it's anti-cancer effect in folk medicine and deserved to be further studied by modern scientific methods.This research aimed to explore its anti-cancer effect and to study the influence on the immunity of melanoma in mice. Methods :Inhibition of different ingredients and concentrations of Wuxing soup on the growth of the mouse melanoma B16 cells was detected by MTT in vitro.Animal experiment was performed to determine its anti-cancer effect in vivo.C57/BL6 mice were randomly divided into three groups after vaccination of B16 cell,and then given intragastrically with different soups for 25 days.All mice were killed on day 26 after inoculation.The weight of tumors were recorded.The anti-tumor immune function was measured by T lymphocyte transforming assay and NK killing assay.The different effect of different ingredients was also observed. Results :Our result showed that different ingredients soup exerted different inhibition on B16 cell growth and Wuxing soup was the strongest one of all and in dose-dependent manner in vitro.The animal experiment indicated that different ingredients soup has different inhibition on melanoma,the soup-treated mouse display improved T lymphocyte transforming assay and NK killing ability.Among the different soups,Wuxing soup showed the strongest anti-cancer effect and immune enhancement. Conclusions :Wuxing soup is an effective anti-cancer agent in melanoma mice and can enhance the immunity of the mice with melanoma.