Objective To optimize the formulations of venlafaxine hydrochloride ( VH ) emulsions by using central composite design-response surface methodology. Methods The effect of amounts of Arlacel P135, VH, PEG-400, and NaCl on the emulsion viscosity, centrifugation breakage, and mean diameter was systemically investigated, respectively. The desirable formulation that combining these three response variables was constructed. Linear equations and a second-order polynomial equation were fitted to the data, and the outcome equation was used to predict the responses in the optimal region. Results There was a quantitative relationship between 4 factors and 3 evaluation indexes and evaluation the “desirability” . The optimal formulation of the VH emulsion were as follows:taking 0. 48 g of Arlacel P135, 0. 40 g of VH, 0. 26 g of PEG-400, and 0. 025 g of NaCl. The experimental values of the response variables were highly closed to the predict values. Conclusion The model presents good prediction and can be used to optimize the preparation of VH emulsion, which obtaining stable W/O emulsion.

BACKGROUND:Activation of Notch signaling plays a critical role in stem cel differentiation, and this effect seems to be cel-type dependent. Little is reported on the role of activation of Notch1 signaling in the differentiation of c-Kit+ bone marrow mesenchymal stem cels. OBJECTIVE:To analyze the influence of activation of Notch1 signaling on the differentiation of c-Kit+ bone marrow mesenchymal stem cels. METHODS:The Notch1 intracelular domain (N1-ICD) was obtained from the cDNA library by PCR and cloned intoBamHI/AgeI digested adenoviral GV314 plasmid to construct N1-ICD overexpressing shuttle plasmid, and the positive clones were verified by sequencing. N1-ICD shuttle plasmid and helper plasmids pBHGloxΔE1,3 Cre were used to co-transfect HEK293T cels to obtain N1-ICD overexpressing adenoviral particles (N1-ICD-Ad). The c-Kit+ subpopulation were isolated from bone marrow mesenchymal stem cels of the Sprague-Dawley rat femurviamagnetic activated cel sorting. After transfection of the c-Kit+ BMSCs with N1-ICD-Ad adenovirus, we assessed the activation of Notch1 signaling and differentiation in c-Kit+ bone marrow mesenchymal stem cels by quantitative RT-PCR and immunofluorescent staining. RESULTS AND CONCLUSION:N1-ICD coding sequence was successfuly generated from the cDNA library, and then was cloned into the linearized adenoviral vectors GV314. The resistant clones were verified by sequencing. With the assistance of packaging plasmids, recombinant N1-ICD-Ad adenovirus plasmids were successful packaged in HEK293T cels, and its title was 2×1012 PFU/L. c-Kit+ bone marrow mesenchymal stem cels with the purity of 91.6% were successfuly isolated from the bone marrow mesenchymal stem cels of the Sprague-Dawley rat femur. Compared with the blank and negative controls, N1-ICD-Ad infection in the c-Kit+ bone marrow mesenchymal stem cels led to substantial accumulation of N1-ICD in the cytoplasm and nuclei, significantly unregulated expressions of Hes1 (a downstream gene of Notch) and cardiomyocyte differentiation genes Nkx2.5 and cTnT, significantly increased the expression of von Wilebrand factor, an endothelial cel differentiation gene, and mildly increased the expression of smooth muscle22α, a smooth muscle cel differentiation gene. These experimental results indicate that the activation of Notch1 signaling contributes to multi-lineages differentiation of c-Kit+ bone marrow mesenchymal stem cels, and the construction of N1-ICD overexpressing adenoviral vector makes the foundation for further research on the role of Notch1 signaling in stem cel biology.

Objective To investigate the clinical effect of percutaneous transluminal angiography in diabetic infrapopliteal arterial disease patients and the influence of post-procedural intraluminal small dose urokinase infusion on infrapopliteal arterial blood flow.Methods From January 2011 to September 2013,37 limbs (16 left and 21 right) in 28 diabetic patients inflicted with infrapopliteal critical limb ischemia underwent endovascular recannalization at our institution and were retrospectively analyzed.Stenotic or occlusive lesions were demonstrated in 74 infrapopliteal vessels,including 30 anterior tibial arteries (ATA),22 posterior tibial arteries (PTA),and 22 peroneal arteries (PA).In 30 limbs,tandem lesions in iliac-femoral arteries were also diagnosed.Antegrade ipsilateral femoral access,retrograde contralateral femoral or brachial arterial access had all been adopted as well as both angioplasty and stenting.Case specific decisions were made based on pre-procedural computed tomographic angiogram (CTA).Ankle-brachial index (ABI) was recorded before and after each procedure.Urokinase was continuously infused through arterial sheath catheter into vessels of target limb from a microinfusion pump at 200 000 to 300 000 units per 24 hour for 48 hours after procedure.Angiogram was performed before and after thrombolysis therapy aiming to ascertain the number of frames of images obtained during the period of time it took blood flow to carry contrast medium from the level of tibial plateau to ankle,which was recorded as index frame count (IFC).Patients were followed up for at least 3 months.ABI and ultrasound or CTA were performed on each follow-up visit to validate patency.Quantitative data such as ABI value and IFC were analyzed using paired samples t-test.Results Thirty two limbs were radiographically recanalized by angioplasty or stenting.Technical success rate was 86.4％ (32/37).Average ABI of all limbs increased significantly from 0.70±0.31 to 0.90± 0.21 (t=10.734,P＜0.05).Of the 32 limbs recanalized,IFC decreased significantly from 6.3 ± 1.6 before thrombolysis to 4.7± 1.4 after thrombolysis (t=12.136,P＜0.05).Six rest pain patients reported significantly alleviated symptoms.Fourteen limbs presented with feet ulcers or gangrene.Of these patients after endovascular treatment,1 underwent ankle level amputation,3 underwent toe amputation and 3 patients who did not seek further treatment reported spontaneous autoamputation and wound healing.The remaining 9 patients reported wound healing within 1 to 3 months.Secondary angioplasty was needed for symptom recurrence in 3 limbs of 3 patients 3 to 24 months after first procedure.Conclusions Endovascular treatment of diabetic infrapopliteal arterial diseases exhibited significant short term effect and was safe to perform.Small dose urokinase infusion after recanalization procedure was safe and effective in helping to improve infragenicular blood flow.

Objective To investigate the nursing of traumatic carotid-cavernous fistula(TCCF)by intervention.Methods 18 patients with TCCF by intervention were given nursing measures including psychological preparation,eye care,disease observation and complications care.Results All patients gained successful embolization,and the symptoms of proptosis and vascular murmur in patients were alleviated.One case received spring embolism because of balloon rupture.The symptoms of vascular spasm in two patients were alleviated with timely treatment.Hyperperfusion syndrome was relieved after lowering blood pressure in two patients.Conclusions The intervention embolization is an effective treatment of TCCF.Good nursing could improve the efficacy and reduce the incidence of complications.

Objective To study the correlation between expression level of als3 gene and the in vivo biofilm formation of Candida albicans in mice.Methods The real-time polymerase chain reaction (PCR) assay was used to detect als3 gene expressions of the clinical Candida albicans isolates from February 2016 to August 2016 in Tianjing No.1 Central Hospital.According to the expression levels of als3 gene, Candida albicans isolates were divided into high and low-expression groups.Thirty C57 mice were randomly assigned to high-expression group (n=15), low-expression group (n=5) and blank group (n=5).Animal model of Candida albicans biofilm was established based on venous catheter and intraperitoneal injection of Candida albicans.Catheters were removed after two weeks;inverted microscope was used for the observation of Candida albicans biofilm formation and transmission electron microscope was used for the observation of its ultrastructure.After irrigating the catheter, the growth of Candida albicans was observed;real-time PCR was used to detect the expression levels of als3 gene 12, 24, and 48 h after the catheter being removed.In this study, t test was used for measurement data and chi-square test was used for rate comparisons.Results In high-expression group, 11 strains (11/15) formed biofilms.In als3 low-expression group, only one strain (1/10) formed biofilm.The difference between these two group was statistically significant (x2=9.64,P<0.05).In als3 high-expression group, two mice died and 8 strains (8/13) formed biofilms, while in low-expression group, there were only 2 strains (2/10) formed biofilms.The difference between these two group was statistically significant (x2=4.02,P<0.05).Thickened Candida albicans membranes and increased mitochondria in high-expression group were observed under transmission electron microscope.In als3 high-expression group, 9 of 13 catheter cultures were positive.However, in als3 low-expression group, 5 of 10 catheter cultures were positive.The difference between these two group was not statistically significant (x2=0.99, P>0.05).In the als3 high-expression group, the expression of als3 gene declined gradually during the biofilm formation.In the als3 low-expression group, the change of als3 gene expression was not obvious.The expressions of als3 gene over time between two groups were significantly different (t=8.7, 10.3 and 9.2, respectively, all P<0.05).Conclusion The high expression of als3 gene in Candida albicans facilitates the formation of biofilm in vivo.

Objective:To observe the efficacy and safety of individual dose of intravitreal conbercept (IVC) in the treatment of retinopathy of prematurity (ROP) before type 1 threshold.Methods:A retrospective case study. From January to July, 2019, 23 cases (46 eyes) of children with type 1 pre-threshold ROP were included in the study. Among them, 14 cases (28 eyes) were male and 9 cases (18 eyes) were female. The mean gestational age at birth was 28.06±1.73 weeks. The average birth weight was 1.14±0.19 kg. The mean corrected gestational age was 34.38±1.41 weeks at the time of first intravitreal injection of IVC. The axial length (AL) of children was measured by A-mode ultrasound before IVC for the first time. According to the calculation of AL, the corresponding injection dose range was 14.23-16.19, 16.20-17.57, 17.58-18.63 mm and the injection dose of IVC was 0.015, 0.020, 0.025 ml (including IVC was 0.15, 0.20, 0.25 mg, respectively). The first IVC dose was 0.015 ml. On the first day before IVC and on the first and seventh days after IVC, 2 ml of arterial blood was taken from children, serum vascular endothelial growth factor (VEGF) concentration was detected. The follow-up time after treatment was ≥1 year. After one year of follow-up, the effective rate and recurrence rate of IVC for the first time were tested by χ2 tests. The short-term changes of injection times, injection intervals, retinal vascularization time and serum VEGF concentration in children were tested by t test. Results:Retinal neovascularization subsided and vascular buckling decreased in all eyes. Iris neovascularization subsided, 1-3 weeks after IVC for the first time. Within one year after the first IVC, 16 eyes underwent IVC twice with or without new blood vessels at the junction of the vascular area. The average corrected gestational age was 40.56±3.81 weeks. The injection dose of IVC was 0.015 ml and 0.020 ml for 2 eyes and 14 eyes, respectively.The mean interval from IVC for the first time was 40.89±8.99 days. Of the 16 eyes who underwent IVC twice, 8 eyes showed neovascularization again in the retinal area with or without blood vessels. The average corrected gestational age was 43.00±1.41 weeks. The injection dose of IVC was 0.020 ml and 0.025 ml for 3 eyes and 5 eyes, respectively. The mean interval of the second IVC was 28.60±6.07 days. The mean interval from the first IVC was 69.20±12.40 days. At the end of follow-up, all eyes were treated effectively (100%, 46/46). The mean time of retinal vascularization was 46.31±3.42 weeks. The average number of injections was 1.52±0.76. On the first day before IVC and on the first and seventh days after IVC, the average serum VEGF concentrations were 111.21±148.71, 25.60±27.71 and 42.99±38.01 pg/ml, respectively. Serum VEGF concentration was significantly lower than that before IVC on the 1st and 7th day after IVC ( Z=-4.054, -2.779; P<0.05). Serum VEGF concentration was higher 7 days after IVC than 1 day after IVC, and the difference was statistically significant ( Z=-2.505, P<0.05). All eyes were not treated by laser photocoagulation or vitrectomy. No eye complications such as lens opacification, endophthalmitis and retinal detachment related to drugs or treatment methods were found in all patients. Conclusion:Intravitreal injection of individualized dose of IVC is effective in the treatment of type 1 pre-threshold ROP. Seven days after treatment, serum VEGF concentration of patients’serum decreases.

Objective To investigate the diagnosis and interventional therapeutic technology for the obstruction of hepatic vein(HV)or inferior vena cava(IVC)after liver transplantation.Methods In the 831 patients who received orthotopic liver transplantation(OLT)and 26 patients who received living donor liver transplantation(LDLT),11 cases were confirmed with HV or IVC obstruction by venography and received interventional treatment from 2 to 111 days after liver transplantation.Of the 11 patients,five had the obstruction of HV anastomosis,five had the obstruction of IVC anastomosis,and one had the obstruction of HV and IVC anastomosis.In the eleven patients,five patients underwent OLT,four patients underwent LDLT,and two pediatric patients underwent reduced-size OLT.Before interventional treatment,9 patients received CT enhanced scans,2 received MR enhanced scans.Follow-up evaluations included liver or renal function tests,clinical symptom,and monitoring of HV or IVC flow.Pressure gradients before and after interventional treatment were compared by using a paired t test.The imaging data and interventional therapeutic technology of 11 cases were retrospectively analyzed.Results In all 11 patients,CT or MRI could clearly show congested areas of the liver,and the location and degree of HV or IVC obstruction.Of the 11 patients,four with HV obstruction and five with IVC obstruction were treated with stent placement,one with HV obstruction was treated with percutaneous transluminal angioplasty(PTA),one with HV and IVC obstruction was treated with HV PTA and IVC stent placement.Interventional technical success was achieved in all patients.The venous pressure gradient across obstruction was significantly reduced from(16.5±4.1)mm Hg(1 mm Hg=0.133 kPa)before the procedure to(2.9±1.7)mm Hg after the rocedure(t= 11.5,P＜0.01).Clinical improvement was noted in 10 patients except one pediatric patient who died of multiple-organs failure at the 9 th day after the treatment During the follow-up period of 9 to 672 days,two patients with PTA treatment had recurrent HV stenosis within one month after treatment,no patient with stent placement developed venous restenosis.No major complications occurred during the procedures.Conclusions Stent placement is safe and effective for HV or IVC obstruction after liver transplantation.CT or MRI before treatment is of important value for the diagnosis of congested areas of the liver,and the observation of HV or IVC obstruction.

To investigate the anti-vasculature effects and the anti-glioma effects of attenuated Salmonella typhimurium vaccine strain expressing VEGFR2 (flk-1) gene, plasmid pcDNA3.1-flk1 was constructed and electro-transfected into live attenuated Salmonella typhimurium strain SL7207. Mouse models of intracranial G1261 glioblastoma were treated with an orally administered attenuated Salmonella typhimurium expressing flk-1 gene. The survival period was recorded and vessel density was observed by immunofluorescence. CTLs activity was measured by MTT assay. Our results showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could significantly inhibit glioblastoma growth, reduce vessel density, prolong the survival period and improve the survival rate in these mice. The flk-1 specific CTLs activity was increased obviously after the vaccination. Our study showed that attenuated Salmonella typhimurium vaccine strain expressing flk-1 gene could break peripheral immune tolerance a in glioma gainst this self-antigen and kill endothelial cells by the orally administered vaccine and can be used for both prophylactic and therapeutic purposes.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Brain Neoplasms ; blood supply ; therapy ; Female ; Glioma ; blood supply ; therapy ; Mice ; Mice, Inbred C57BL ; Neovascularization, Pathologic ; Salmonella Vaccines ; immunology ; pharmacology ; Salmonella typhimurium ; immunology ; Transformation, Genetic ; Vaccines, Attenuated ; genetics ; immunology ; pharmacology ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology

Objective To compare the efficacy of percutaneous and endoscopic treatment for the biliary stricture(BS) after liver transplantation (LT).Methods The result of percutaneous transhepatic cholangiography (PTC) and drainage ( PTC group) and endoscopic retrograde cholangiopancreatography (ERCP group) for the BS in 132 post-LT patients were analyzed retrospectively.Ninety-nine patients received PTC treatment,and 59 patients received ERCP treatment,26 patients converted to PTC treatment because of the poor efficacy or failure of the ERCP treatment.The operation success rate,complication rate,cure rate and remission rate of the two groups were compared with X2 test.Results The BS types of PTC and ERCP group were different significantly( P ＜ 0.01 ),with more non-anostomotic stricture in PTC group and more anostomotic stricture in ERCP group.The operation success rate of PTC group was higher than of ERCP group( 100％ vs 97％ ) (P ＜0.01 ),and the complication rate of PTC group was lower than of ERCP group.The overall cure and remission rate of PTC and ERCP group were not different significantly(32.3％ vs 45.8％,94.9％ vs 88.1％ ) (P ＞0.05).The cure and remission rate of PTC and ERCP treatment for each subtype of BS were not different significantly ( P ＞ 0.05 ).Conclusions The efficacy of PTC treatment for the post-LT BS is equivalent to that of ERCP treatment.PTC can be considered the first-line option for the post-LT BS.

Objective To investigate the clinical effects of therapeutic hypothermia on the mortality and neurological function recovery in patients with acute large hemispheric infarction (LHI).Methods Seventy-three acute LHI patients (within 24 hours of symptom onset) admitted to our hospital from February 2015 to July 2016 were randomized to the hypothermia group (37 patients)and the control group (36 patients).Patients in the hypothermia group were given standard medical treatment and the surface cooling with a target temperature between 32-35 ℃ lasting for 5-7 days.Patients in the control group were given standard medical treatment and maintained a target temperature of normothermia.All patients were observed the primary end points including mortality and the modified Rankin Scale (mRS) score at 3 months.Results At three months,nine patients of survivors in the hypothermia group had good outcome (mRS score 0-3),whereas only five patients of survivors in the control group,though there was no significant difference (9/17 vs 5/19,x2 =2.676,P＞ 0.05)between the two groups.Twenty patients (54.1％)died in the hypothermia group,while 17 patients(47.2％) in the control group(x2 =0.341,P ＞0.05).Conclusion Therapeutic hypothermia is safe and feasible to patients with acute LHI,but may neither reduce mortality nor improve the neurological outcome in survivors.