Objective To prepare recombinant human papillomavirus 58 L1 protein virus-like particles(HPV58 LI VLPs)in Pichia pastoris and immunize mice with the purified product to evaluate the immunogenicity of HPV58 L1 VLPs.Methods The HPV58 L1 multi-copy expression plasmid was constructed and transformed into Pichia pastoris KM71.The highly expressed Pichia pastoris engineering strain was screened and cultured in 5 L fermenter.The expressed HPV58 L1 VLPs were isolated and purified,which were then used to immunize 80 female BALB/c mice at different doses by intraperitoneal injection.After four weeks,the immune effect was evaluated by median effective dose(ED_(50)).Results Restriction analysis results showed that HPV58 L1 expression vectors were constructed correctly.Western blot,amino acid sequencing and transmission electron microscopy(TEM) results showed that HPV58 L1 protein was correctly expressed in Pichia pastoris and could self-assemble into VLPs at a diameter of about 50 nm.After purification by ion exchange and size exclusion chromatography,the HPV58 L1 VLPs reached a purity above 95%,and the ED_(50) in mouse model was 0.009 μg.Conclusion Recombinant HPV58 L1 VLPs with good immunogenicity were successfully prepared by using Pichia pastoris.

Objective To investigate the effect of two kinds of drainage operation including incision and drainage of abdominal operation and percutaneous puncture and drainage operation under ultrasound guidance on the short-term curative effect, mortality and complications risk in patients with single giant bacterial liver abscess. Methods Eighty patients with single giant bacterial liver abscess were selected, and all patients were treated with conventional therapy. Among them, 40 patients underwent incision and drainage of abdominal operation (control group), and 40 patients underwent percutaneous puncture and drainage operation under ultrasound guidance (observation group). The short-term curative effect, operation time, time of returning to normal body temperature, time of returning to normal white blood cell, hospitalization time, mortality and incidence of postoperative complication were compared between 2 groups. Results There were no statistical differences in total effective rate and mortality between control group and observation group: 92.5%(37/40) vs. 97.5%(39/40) and 5.0%(2/40) vs. 7.5%(3/40), P>0.05. The operation time, time of returning to normal body temperature, time of returning to normal white blood cell and hospitalization time in observation group were significantly lower than those in control group:(44.52 ± 5.68) min vs. (98.59 ± 12.70) min, (3.25 ± 0.74) d vs. (4.86 ± 1.32) d, (5.20 ± 1.35) d vs. (7.42 ± 1.79) d, (16.94 ± 3.61) d vs. (25.52 ± 5.28) d. The incidence of postoperative complication was significantly lower than that in control group:10.0%(4/40) vs. 27.5%(11/40), and there were statistical differences (P<0.05). Conclusions Two kinds of drainage operation in the treatment in patients with single giant bacterial liver abscess possesses the same clinical efficacy for short-and long-term curative effect; but percutaneous puncture and drainage operation under ultrasound guidance application can efficiently alleviate the clinical symptoms and signs, accelerate the recovery process of disease and be helpful to reduce the risk of postoperative complications.

Objective To study the value of video EEG combined with MRI in the diagnosis of intractable epilepsy.Methods40 patients with intractable epilepsy treated in our hospital from 2013 to 2015 were selected as the subjects.In 40 patients, 26 patients with hippocampal sclerosis, 2 cases of patients with temporal lobe atrophy with cerebral dysplasia, 4 cases of patients with cerebral gray matter and macrogyria, 3 cases of vascular malformation, 1 cases of patients with intracerebral cyst, 2 cases of patients with glioma, 2 cases of patients with traumatic epilepsy.Retrospective of The clinical data of the selected patients were analyzed and the manifestations of epileptic foci were reflected by MRI, The use of video EEG was used to detect the status and source of EEG during epileptic seizure and epilepsy Analysis, combined with MRI scan and video EEG results, control surgery pathological changes, analysis of the results, draw conclusions.ResultsThere were 21 cases of the accurate location of interictal EEG.The accuracy was 52.5%, and the accuracy of the location of epileptogenic foci was about 40, and the accuracy was about 100%.The accuracy of MRI in the diagnosis of epileptic foci and related lesions was 35 cases, the accuracy rate was 87.5%, and the number of lesions was qualitative and accurate in the case of 30 cases, with an accuracy rate of 75%.ConclusionVideo EEG combined with MRI examination for drug refractory epilepsy diagnosis has important clinical value, can make a more accurate diagnosis of patients with epileptogenic foci, provide information for surgical treatment of intractable epilepsy patients, with further clinical promotion and application significance.

Objective To explore the influence of combination anaesthesia application of midazolam and propofol on stress reactions in radical resection of colon cancer,and to explore novel clinical compatibility of anaesthesia drugs.Methods 40 patients received radical resection of colon cancer were selected and divided into control group and combination group randomly and evenly.Control group received general anesthesia by propofol,while combination group received additional midazolam when anesthesia induction was taken.Glycemia,CRP,complement C3 and cortisol were observed and compared at T1 (before anesthesia induction),T2 (4h after beginning of surgery) and T3 (24h after surgery).Results Compared with T1,glu increased significantly at T2 and T3 (all P ＜ 0.05) ; Compared with combination group,glycemia increased more significant in control group (P ＜ 0.05) ; CRP and cortisol increased significantly in all patients compared with T1 (P ＜ 0.05),more significant in control group (P ＜ 0.05) ; a significant decrease after increase trend was identified for complement C3 (P ＜ 0.05),more significant decrease was found in control group at T2 (P ＜ 0.05).Conclusion Combination anesthesia of propofol and midazolam could decrease the strength of stress reaction in radical resection of colon cancer.

Objective To compare the clinical efficacy of tension-free hernioplasty with three operative methods, and analyze the reasons of complications. Methods All Chinese articles dealing with the tension-free hernioplasty included in CNKI from 1997 to 2004 were collected, and divided into three groups according to different operative methods, the clinical efficacy of which was compared. Results The frequecy of foreign matter feel in the Rutknow group was significantly higher than that in the other two groups (P

Objective To evaluate the value of multiplanar reconstruction (MPR) of spiral CT in diagnosing and staging of cervical cancer .Methods SCT scan with a 3～5 mm collimation thickness and MPR in 1.5～2.5 mm reconstruction interval were performed in 20 patients with cervical cancer , confirmed by biopsy pathology. Results The sagittal imaging could show entirely the growing character of a carcinomal mass;the coronal and transverse imagings showed clearly parametrial invasion and pelvic lymph node metastases. Conclusion MPR images in the pelvic could show entirely the character of growing and invasion of a carcinomal mass. It is an important value in dignosing and staging of cervical cancer.

[ ABSTRACT] AIM:To establish a perforated whole-cell patch-clamp technique withβ-escin to record L-type cal-cium current (ICa,L) in osteoblasts.METHODS:ROS 17/2.8 is a rat osteoblast-like osteosarcoma cell line.β-escin was applied to the pipette solution to permeabilize the cell membrane and the perforated patch recording mode was obtained. RESULTS:β-escin at concentration of 50μmol/L easily permeabilized the cell membrane and obtained a perforated patch recording mode in 2~7 min.This technique prevented ICa,L rundown and preserved cytoplasmic signaling pathways.CON-CLUSION:β-escin may be used as an alternative ionophore for perforated patch-clamp studies in osteoblasts and results in minimal rundown that could facilitate recordings of ICa,L in osteoblasts.

The combination of high-speed countercurrent chromatography and rapid bioactive assay is a desirable method in the screening of bioactive components from CMM and natural products.This review focused on the application status of high-speed countercurrent chromatography to the rapid screening of bioactive components from CMM and natural products based on our previous studies and the recent progress over it.

Objective This study intends to evaluate different acoustic windows that can be used to obtain the midsagittal plane of fetal brain during abdominal ultrasound scanning and the application of acoustic windows in practice.Methods The midsagittal planes were obtained respectively through anterior fontanel,posterior fontanel,sagittal suture and forehead seam of 12 aborted fetuses,and feasible acoustic windows were used as standard acoustic windows to obtain midsagittal planes of second trimester screening fetuses,which were divided into two groups according to the position of fetal brain,cephalic presentation group (367 cases) and non-cephalic presentation group (384 cases).The success rates were compared between groups.Results Of the second trimester fetuses,497 midsagittal planes of 751 cases were successfully obtained.The success rate of the cephalic presentation fetuses (32.7％,120/367) was statistically lower than that of the non-cephalic presentation fetuses (98.2％,377/384,P ＜ 0.01).Of the cephalic presentation fetuses,the combined acoustic windows acquisition success rate of the forehead,anterior fontanelle,sagittal suture and posterior fontanel acoustic window was 32.7％ (32.7％,120/367),significantly higher than the single acquisition success rate of the forehead,anterior fontanelle,sagittal suture and posterior fontanel acoustic window (17.2％,25.9％,1.6％ and 7.6％,all P ＜ 0.01).Of the non-cephalic presentation fetuses,the combined acoustic windows acquisition success rate of the forehead,sagittal suture and posterior fontanel acoustic window was 98.2％ (98.2％,377/384),significantly higher than the single acquisition success rate of the forehead,anterior fontanelle,sagittal suture and posterior fontanel acoustic window (31.8％,61.2％,64.8％ and 62.0％,all P ＜ 0.01).Conclusions The midsagittal plane of fetal brain can be obtained through forehead seam,anterior fontanel,posterior fontanel and sagittal suture acoustic windows in transabdominal screening.The combination of four acoustic windows can increase the success rate of obtaining midsagittal planes.

Objective To investigate the influence of combined irbesartan/hydrochlomthiazide on morning blood pressure surge(MBPS)and left ventricular mass(LVM) in essential hypertension.Methods After 4-8 weeks' run-in period,if blood pressure wagn't up to the standard,the 120 patients were given irbesartan/hydrochlorothiazide once daily from 150 mg/12.5 mg for 12 months to 300 mg/25mg for another 12 months.MBPS was conducted by arnbuhry blood pressure monitoring(ABPM)and LVM was determined by echocardiography at the end of placebo baseline and 12 months later.Results (1)After treatment the patients with MBPS(+)decreased and patients with MBPS(-) increased.(2)After treatment the extents of MBPS were reduced in MBPS(+)patients[systolic blood pressure(SBP)difference(16.1±1.8)mm Hg(1mm Hg=0.133 kPa)vs(29.4±2.8)mm Hg,diastolic blood pressure(DBP)difference(10.2±2.3)mm Hg vs(21.2±2.2)mm Hg,P<0.01]with little change in MBPS(-)patients[SBP difference(11.2±2.4)mm Hg vs(10.1±1.2)mm Hg,DBP difference(5.9±1.9)mm Hg vs(6.8±3.2)mm Hg]compared with before treatment.(3)LVM indexes of all patients were significantly reduced.Conclusion Irbesartan/hydrochlorothiazide can effectively attenuate the extent of MBPS, and has advantages on reversing left ventricular hypertrophy.