Objective To investigate the clinical efficacy of three-dimensional (3D) and two-dimensional (2D) laparoscopic surgeries in the treatment of Todani type Ⅰ choledochal cyst.Methods The retrospective cohort study was conducted.The clinical data of 59 patients with Todani type Ⅰ choledochal cyst who were admitted to the People's Hospital of Hunan Province between January 2013 and January 2016 were collected.Thirty patients undergoing 2D laparoscopic surgery between January 2013 and June 2014 were allocated into the 2D group and 29 patients undergoing 3D laparoscopic surgery between July 2014 and January 2016 were allocated into the 3D group.There were the same Trocar placement and surgical procedure in the 2 groups,and surgical procedure completely followed the treatment principle of Todani type Ⅰ choledochal cyst.Observation indicators included (Ⅰ) surgical situations:conversion to open surgery,operation time,volume of intraoperative blood loss,(2) postoperative situations:postoperative complications,(3) follow-up.Patients were followed up by outpatient examination or telephone interview to detect postoperative recovery up to April 30,2016.Measurement data with skewed distribution were presented as M (range) and analyzed using the Mann-Whitney U test.Count data were compared by Fisher exact probability.Results (1) Surgical situations:patients in the 2 groups underwent laparoscopic choledochal cystectomy + Roux-en-Y hepaticojejunostomy.Two patients in the 2D group received conversion to open surgery and patients in the 3D group received the successful surgery without conversion to open surgery.Rate of conversion to open surgery in the 2D and 3D groups were 6.7％ (2/30) and 0,respectively,with no statistically significant difference (P ＞ 0.05).Operation time in the 2D and 3D groups were 285 minutes (range,240-390 minutes) and 190 minutes (range,140-215 minutes),with a statistically significant difference (U =40.0,P ＜ 0.05).Volume of intraoperative blood loss in the 2D and 3D groups were 50 mL (range,10-300mL) and 45 mL (range,20-250 mL),with no statistically significant difference (U =1 018.5,P ＞ 0.05).(2)Postoperative situations:patients in the 2 groups had good recovery,without occurrence of severe complications in Clavien-Dindo≥ Ⅲ stage.Four and 1 patients in the 2D and 3D groups were complicated with bile leakage (in Ⅱ stage of Clavien-Dindo) and 1 and 1 were complicated with upper gastrointestinal hemorrhage (in]][stage of Clavien-Dindo),respectively,with no statistically significant difference (P ＞ 0.05).Overall incidence of complications in the 2D and 3D groups were 16.7％ (5/30) and 10.3％ (3/29),with no statistically significant difference (P ＞ 0.05).All the patients were cured by conservative treatment.(3) Follow-up:59 patients were followed up for 5-36 months,with good recovery and without occurrence of reflux cholangitis,hepatic and intestinal anastomosis stenosis and reoperation.Conclusions 3D and 2D laparoscopic surgeries are safe and effective for Todani type Ⅰ choledochal cyst.Compared with 2D laparoscopic surgery,3D laparoscopic surgery can reduce the operation time and not increase the complications,and it should be discreetly promoted based on the experiences of surgeons.

The M and NP genes of H5N1 avian influenza virus (A/chicken/Hubei/489/2004) were amplified by RT-PCR from viral RNA,and cloned into pMD 18-T vector respectively.The expression plasmid containing the M gene (pHM6-m) or the NP gene (pHM6-np) was then constructed by inserting the M or NP gene into the pHM6 eukaryote expression vector; the constructed plasmid was then sequenced.32 BALB/c mice (6-week-old) were divided into four groups at random.Three groups of BALB/c mice were inoculated one time the intramuscular route with either 30 μg of plasmid pHM6-m,30 μg of plasmid pHM6-np or the mixture of plasmid pHM6-m (15 μg ) and pHM6-np(15 μg) respectively.A additional group of mice were injected with 100 μ1 PBS as controls.Two weeks later,all mice were challenged with homologous H5N1 avian influenza virus,and observed in the following 12 days.The survival rates of mice in the pHM6-m group,the pHM6-np group and mixed plasmids group were 62.5% ,25.0% and 50.0%,respectively.Results showed that effective protection could be provided by either pHM6-m or pHM6-np,but pHM6-m provided a better protective effect than pHM6-np.

AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

Adult ; Bone Cysts, Aneurysmal ; diagnosis ; surgery ; Bone Neoplasms ; diagnosis ; surgery ; Chondroblastoma ; diagnosis ; surgery ; Female ; Humans ; Male ; Talus ; surgery ; Young Adult

Objective To investigate the effect of hosphocreatine therapy on the plasma cardiotrophin-1(CT-1) and N terminal probrain natriuretic (NT-proBNP) in elderly hypertensive patients with heart failure. Methods A total of 76 hy-pertensive patients with heart failure, aged 65 or over, were randomly divided into treatment group and control group (n=38 for each group). The control group received routine anti-heart failure treatment. The treatment group received conventional therapy plus creatine phosphate sodium for 2 weeks. The plasma levels of CT-1 and NT-proBNP were determined in two groups. The plasma CT-1 level was measured by a double antibody sandwich enzyme-linked immunosorbent assay (ELISA). The plasma level of NT-proBNP was tested by Rui Pu fluorescent dry quantitative analyzer. Results The plasma levels of CT-1 and NT-proBNP were significantly lower after treatment in two groups (P＜0.01). The plasma levels of CT-1 and NT-proBNP were significantly decreased in treatment group than those in control group (P＜0.05). The total effective rate was 89.47%in treatment group, which was significantly higher than that of control group (71.05%, P＜0.05). Symptoms of heart failure improved in one week (21 cases in treatment group/9 cases in control group) and in two weeks (13 cases in treatment group/18 cases in control group). Conclusion The conventional therapy plus creatine phosphate sodium can decrease the plasma CT-1 and NT-proBNP levels in elderly hypertensive patients with heart failure, which improves symptoms of heart failure in a shorter period of time.

Objective The aim of this study is to detect the mRNA and protein expression levels of ERCC1 and XPF genes among different age groups of healthy Chinese Han individuals,and to analyze the correlation between the mRNA and protein expression levels andthe age of individuals in order to find new molecular markers for forensic age estimation.Methods Peripheral blood samples were obtained from 150 unrelated healthy Chinese Han individuals.The plasma was centrifuged from the whole blood by gradient centrifugation,and the totalRNA was extractedwithTrizol fromperipheral blood mononuclear cells(PBMCs).Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to quantitatively analyze the mRNA relative expression levels of ERCC1and XPF in PBMCs.Enzyme linked immunosorbent assay (ELISA) was used to quantitatively analyze the protein expression levels of ERCC1and XPF in plasma.Results There were no significant differences in the mRNA relative expression levels of ERCC1 and XPF in PBMCs between males and females(P＞0.05).Significant differences were found in the mRNA relative expression levels of ERCC1 and XPF between different age groups (P＜0.05).Regression analysis showed thatthe mRNA relative expression levels of ERCC1 and XPF were both negatively correlated with age.The correlation coefficients(r) were-0.578 and-0.844,respectively.When the age was used as independent variable(x) and the mRNA expression relative level as dependent variable (y),the fitting curveswere Y=3.3E-5X2-0.0261X+1.9175 (R2=0.3244,P＜0.01),Y=0.0003X2-0.0459X+2.0439 R2=0.729,P＜0.01),respectively.There were no significant differences inthe protein expression levels of ERCC1 and XPF in plasma between different age groups or genders (P＞0.05).Conclusion The mRNA relative expression levels of ERCC1 and XPF in PBMCsdeclined with the increase of age,however,the protein expression levels in plasma were unrelated to age.ERCC 1 and XPF genes can be used asnew molecular markers for forensic age estimation,so as to providetheoretical basis for establishing the mathematical model of ERCC1/XPF genesin concern ofindividual ages.

Objective To establish the cardiac arrest-cardiopulmonary resuscitation model in rats, and to observe the effect of mild hypothermia on autophagy in hippocampal CA1 neurons after ROSC.Methods A total of 36 Wistar rats were randomly divided into two groups: normal temperature treatment group(NT group) and mild hypothermia treatment group(HT group).To establish the cardiac arrest-cardiopulmonary resuscitation(CA-CPR) model in rats by epicardial electrical stimulation induced ventricular fibrillation, and to sacrifice 3 animals in each group to obtain the brain cortex in 2nd and 4th hours after ROSC in order to observe the expression of p-AMPK by electron microscope and LC3 granules through Western blot.The neurological deficit score(NDS) was assessed in 24、48、72 hours respectively after ROSC.To sacrifice the animals so as to take the cerebrum in 72 hours after ROSC, then calculate the apoptotic index of the hippocampal CA1 neurons, which were dyed through TUNEL method.Results The expression of p-AMPK、Beclin-1 and LC3-Ⅱ/LC3-Ⅰratio in Normothermia group were all lower than the Mild hypothermia group(P<0.05), the neurons plasma of hippocampal CA1 area in the Hypothermia group demonstrated obvious LC3 granules formation, the NDS score of the Normothermia group and the Mild hypothermia group in ROSC24h、ROSC48h、ROSC72h were 320vs205、285vs140、266vs120, respectively.The apoptotic index of the hippocampal CA1 area in the Normothemia group in ROSC72h was higher than the Mild hypothermia group,(P<0.05).Conclusions Mild hypothermia after cardiopulmonary resuscitation promotes autophagy of the hippocampal CA1 area neurons in rats and reduce neuronal apoptosis.

Total DNA of B. amyloliquefaciens and S. aureus were isolated and cut partially with Hind Ⅲ Separately, The genes for barnase and for staphylococcal nucl ease were obtained by PCR and cloned into pGEM7Z-f(+), The sequencing show that the n ucleotide (nt) sequence of the barnase is 99.3% homologous with the previously determimed sequence and the deduced animo acid (an)sequence found to correspond precisely to the previously determined sequence; the nucleotide (nt) sequence of the staphylococcal nuclease is 99.1% homologous with the previously determined sequence and the deduced amino acid (an) sequence 99.3% homologous with the previously determinded sequence.These have built basis of making use of special expression or activation of nuclease to produce the male sterility plant by plant gene engineering and to explore new antiviral strategy.

More than 80 strains producing lipases were screened from oily soil of vegetable oil plant, meat processing factory and dairy factory in Jinan city, Shandong Province, which included bacteria, moulds and yeast. Conditions for lipase production and properties of some enzymes were studied. One drug-resistant mutant strain Y-11 with higher lipase activity was from Trichosporon sp. Y-1. In addition, Optimization of lipase production and characterization of enzymes were carried out. On the basis of above experiments, a characteristic library of stains producing lipases was established.

Objective To identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.