ABSTRACT:Objective To construct V-set and immunoglobulin domain containing 4 (Vsig4)nanobodies (Nbs) as specific macrophage probes so as to use them as molecular probes of macrophagocytes.Methods A nanobody phage library was generated by using peripheral blood lymphocytes isolated from an alpaca immunized with recombinant Vsig4 protein.After three rounds of selection against recombinant Vsig4.The Nbs were subjected to sequencing and genome alignment to obtain VHH sequence.Nbs were isolated and tested for Vsig4 specificity in an ELISA using recombinant Vsig4.The affinity capacity of Nbs was verified by the cell line stably expressing Vsig4. Results A nanobody phage library with an estimated 7.27 × 107 clones with 70% insertion was successfully constructed.Totally 1 3 6 Vsig4-positive clones were sequenced and aligned according to different CDR3 sequences. In summary,1 5 Vsig4 nanobodies were obtained and grouped into 3 different CDR3 epitopes.The affinity of representing nanobody and Vsig4 was analyzed via ELISA;Nb1 1 9 showed the highest affinity against both recombinant and native Vsig4.Conclusion We successfully constructed and screened Vsig4 specific nanobody number 1 1 9 with high affinity and specificity.It can help with macrophage detection and in vivo monitoring.

Immunologic classification of acute lymphocytic leukemia (ALL) was carried out by using 11 - 13 kinds of monoclonal antibodies against human leucocyte differentiation antigens in 19 cases. The results showed that of these 19 cases, there were T-ALL in 4, B-ALL in 4, C-ALL in 7, AUL in 2 and hybrid type of ALL (H-ALL) in 2. The 5 types mentioned above could also be classified, according to the immunologic criteria described by Foon in 1986, into 3 major groups: T-ALL, non-T-ALL and H-ALL. The Foon's classification method is useful for judgement of malignant cell source and differentiated stage, and for further understanding the nature of ALL cells.

To solve separating problem of natural soda, fifty-seven strains screened from soil, floul water and activated mud were of flocculating activity. Two strains of bacteria, which were screened from above mentioned strains have higher activity and better steady than the whole culture liquid of bacteria was observed that its flocculating use to natural soda was strong and the mean flocculating rate of two strains were 79.80% and 87.% respectively.

Parkinson's disease (PD) is a degenerative disease of the central nervous system due to the loss or death of dopaminergic neurons in the substantia nigra. Clinically, levodopa is the most effective and commonly used drug for PD treatment. However, long-term levodopa therapy is prone to motor complications and other side effects caused by excessive peripheral dopamine production, which has become an urgent problem to be solved in PD treatment. Dopamine receptor (DR) agonists are similar to dopamine. They can directly stimulate postsynaptic dopamine receptors, produce the same effect as dopamine, delay the application of levodopa as much as possible, and reduce complications caused by long-term use of levodopa. Therefore, screening effective dopamine receptor agonists has become a key issue in the study and treatment of PD. In order to establish a rapid, stable and reliable method for dopamine receptor agonist screening, this study used the human dopamine receptor 2 (DRD2) gene fused with a circular permuted EGFP (cpEGFP) to construct a recombinant gene, packaged with lentiviral vector, and the vector replaced the parted inner transmembrane domain of the third intracellular loop (ICL3) of genetically-encoded GPCR-activation based (GRAB) sensors. The fluorescence of GPCR-fused cpEGFP is regulated by conformational changes mediated by the interaction of dopamine receptor agonists with GPCRs without altering GPCR activity. The HEK293T cells were infected with viral vector, screened by puromycin to select highly expressed cells. Dopamine receptor agonists (including dopamine, bromocriptine mesylate, cabergoline, pramipexole) were used as positive drugs to explore the best screening and detection conditions, establishing a stable model to evaluate the dopamine receptor agonist. The results showed that the optimal filter for the dopamine receptor agonist in this study was the cell seeding count of 7×10⁴, and the effective concentration of the positive drug was 1-100 µmol·L^-1. In addition, pretreated with 10 µmol·L^-1 dopamine receptor antagonists (including chlorprothixol hydrochloride, domperidone, and sulpiride), the positive fluorescence signal of overexpressed DRD2-cpEGFP HEK293T cells could not be detected when exposed to 10 µmol·L^-1 dopamine receptor agonists, which proved that dopamine receptor antagonists could block the activity of dopamine receptor agonists, so they cannot activate dopamine receptor allosteric, indicating that the model has good specificity and can also be used for the screening and detection of new dopamine receptor antagonists. In summary, the study constructs a stable dopamine sensor detection system, which can effectively screen potential dopamine receptor agonists. The operation procedures are simple and rapid. And it can be used for a large-scale screening providing a fundamental methodology for drug development and PD treatment targeted on DRD2.

OBJECTIVETo investigate the changes of vasoconstriction and vasodilatation under different temperature conditions and the protective effects of Vitamin E (Vit E) against endothelial injury induced by hypothermia.

METHODSThe tail arterial rings were prepared for isometric tension recording using multi wire myograph system. The effect of temperature on relaxation and construction was evaluated. Incubate the arterial rings with different concentration of Vit E when they were exposed to hypothermia, then acetylcholine (ACh)-induced endothelium-dependent relaxation was investigated to evaluate the activity of endothelial.

RESULTS(1) The hypothermia could enhanced the dose-dependent construction induced by PE in mice tail artery. (2) Exposure to hypothermia also resulted in increase of sodium nitroprusside (SNP)-induced re-After incubation with Vit E, the vascular relaxation responses to ACh increased in an endothelium-dependent manner, when compared with the hypothermia-treated group.

CONCLUSIONThe vascular function of constriction was attenuated by hypothermia, while the relaxation was increased. Vit E could prevent the hypothermia-induced decrease in vascular endothelial cells.

Animals ; Arteries ; drug effects ; physiology ; Cold Temperature ; Hypothermia ; In Vitro Techniques ; Male ; Mice ; Prazosin ; pharmacology ; Solanaceous Alkaloids ; pharmacology ; Vasoconstriction ; drug effects ; Vasodilation ; drug effects ; Vasodilator Agents ; pharmacology ; Vitamin E ; pharmacology

OBJECTIVETo investigate the advantages and disadvantages of dual energy with dual-source computed tomography (CT) in cerebral angiography by comparing those of neuro-digital subtraction angiography (Neuro-DSA) .

METHODSTotally 300 patients with normal CT angiography (CTA) images underwent dual energy CT angiography (DE-CTA groups, n=200) and Neuro-DSA scan (Neuro-DSA groups, n=100) using dual source CT. The comparison of two scan approaches was based on image quality, radiation does, post-processing methods, and duration of scanning and subtraction.

RESULTSThe image quality of intracranial blood vessels in DE-CTA group showed no significant difference with that in Neuro-DSA group (P>0.05) , while the number of Grade I images was significantly higher than those of other three grades (P<0.01) . However, the effect of the skull base bone removal with Neuro-DSA is superior to DE-CTA (P<0.01) . The duration of scanning was significantly longer in DE-CTA group than in Neuro-DSA group (P<0.01) . However, the radiation dose, time of entire examination, and time of subtraction were significantly lower/shorter in DE-CTA group than in Neuro-DSA group (P<0.01) . The radiation dose of DE-CTA group is 26.3% lower than Neuro-DSA group.

CONCLUSIONSThe image quality of intracranial blood vessels with head DE-CTA is same as Neuro-DSA, while head DE-CTA has lower radiation doses and higher inspection efficiency than Neuro-DSA. However, the effect of the skull base bone removal needs to be improved.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Angiography, Digital Subtraction ; methods ; Cerebral Angiography ; methods ; Female ; Humans ; Image Processing, Computer-Assisted ; Male ; Middle Aged ; Radiation Dosage ; Sensitivity and Specificity ; Tomography, X-Ray Computed ; methods ; Young Adult

To established an effective GC-MS /MS method for the contents determination of the residual DEHP in injection equipment, and investigate the effect of the pretreatment on the measurement. To simulate the clinical conditions of use, under the condition of 37 degrees C balance extraction, extract liquor by chloroform extraction, then the extract followed by analysis of GC-MS /MS. The method was simple, rapid, sensitive and accurate. The limits of quantitation (LOQ, S/N = 5) of cyclohexanone was 0.075 μg/mL, The spiked average recoveries ranged from 92% to 98%. The relative standard deviations (RSDs) of the method ranged from 1.01% to 1.61%, The method was simple, fast, sensitive and accurate, and may serve as a mass control method for residual DEHP in injection equipment.
Cyclohexanones ; chemistry ; Diethylhexyl Phthalate ; chemistry ; Equipment Contamination ; Gas Chromatography-Mass Spectrometry ; Injections ; instrumentation ; Limit of Detection ; Plasticizers ; chemistry

Objective To analyze the diagnosis and prognosis value of abdominal CT scans in patients with acute pancreatitis complications.Methods 151 cases with acute pancreatitis were selected.The relationship between abdominal CT performances and the common complications and death in patients was analyzed.The prognosis of patients with different Balthazar CT severity index (CTSI) grade was compared.Results The complication rate of patients with CT signs of fatty liver, pleural effusion, liver gap effusion, adrenal gland involvement (AGI), penirenal space involvement (PSI) and gastric bare area involvement (GBAI) was significantly higher than that of patients with negative CT findings above (P<0.05).With CTSI grading increasing, the patient''s fasting time, heating time, hospital stay, recovery time of blood amylase were extended, and the incidence of pseudo cyst, transit surgery, organ failure and death was gradually increasing (P<0.05).Conclusion Severe fatty liver, AGI, GBAI, PSI and liver gap effusion are risk factors for acute complications and death in patients with pancreatitis.

Objective To conclude the preliminary results of aortic valve repair with autopericardium.Methods After routine median sternotomy the pericardium was isolated and immersed into the 0.6% glutaraldehyde immersion for 10 minutes, then rinsed off before use.Conventional extracorporeal circulation was established and diseased aortic valves were removed.The curves lengths between three commissures measured with the specific sizers.The corresponding size of the pericardial patchs was used to cut the new aortic leaflets.The continuous Johnson Prolene 4-0 sutures were used to fixed the new aortic valve leaflets to the anulus.Another new 4-0 prolene suture were used to fix the new three commissures outside the aortic wall with mat.The new mat stripes were used to fix the sinus in condition with the aortic sinus dilation or bileaflets malformation.Results Since 2015 October total 27 patients received aortic valve repair with autologous pericardium.The age was in the range of 27 to 60 years old including 18 female and 9 male.There were 7 cases diagnosed as aortic valve stenosis and 20 cases aortic insufficiency.And there were 3 cases bileaflets aortic valve deformity.The mean bypass time is(104±25)mins.Postoperative esophageal ultrasound showed good coaptation of the aortic valve without significant regurgitation.The mean transvalve gradient was(8.5±3.2)mmHg(1 mmHg=0.133 kPa).In the early stage one case diagnosed with aortic valve stenosis and regurgitation received monocusp repaired with autologous pericardial leave.But there was still moderate aortic regurgitation and had to receive the aortic valve replacement.All patients recovered well and were treated with Aspirin for 6 months.The followed up lasted for 3 to 13 months.Mean aortic occlusion time was(104±25) minutes.The patients were followed up for 3-13 months, the cardiac function and aortic valve function were stable, and no gradient increased.Conclusion It is relatively simple to perform the aortic valve repair with individual autologous pericardial sizer technique and the primary clinic results are satisfactory.It is worthy of further study and long-term follow-up.

Objective To compare the mid-term results of video-assisted pulmonary veins isolation and box-lesion for lone atrial fibrillation.Methods Between June 2010 to November 2012,80 lone atrial fibrillation(LAF) patients underwent minimally invasive surgical ablation in Beijing Anzhen Hospital,and all of them received left atrial appendage excision and Marshall ligament break in the surgery.Among them,37 cases were performed two epicardial ablation lines in order to created box-lesion based on PVI(14 paroxysmal AF and 23 persistent AF).Follow-up was finished after discharge.Results Mean follow-up was(18.8 ± 7.4) months,and 2 patients were died with non-cardiac disease.The success rate is 78.8％ (Paroxysmal AF 87.9％;Persistent AF 72.3％).In the paroxysmal AF patients,the success rate for PVI and Box lesion treatment was 84.6％ (11/13) and 92.9％ (13/14),P =0.45;in the persistent AF patients,the success rate for PVI and Box-lesion was 58.3％ (14/24) and 87.5％ (14/16),P =0.04.Conclusion Addition of epicedial ablation lines might increase the cure rate for lone AF therapy,especially for persistent AF.