Objective: To express glycosyl-phosphatidylinositol (GPI) modified Met- RANTES fusion protein on Chinese hamster ovary (CHO) cells and to develop a novel immunosuppressant GPI anchored form of Met-RANTES. Methods: The eukaryotic expression vector PEF/GPI-Met-RANTES were constructed and transfected into CHO cells by electroporation. The transfectants were selected with methotrexate (MTX). Expression of the recombinant protein was assessed by flow cytometric analysis, cell immunofluorescence staining and immunogold electron microscopy. Results: The chimeric molecules of GPI anchored form of Met-RANTES including the whole reading frame were constructed, and the sequence was identical to the designed sequence. GPI anchored form of Met-RANTES was stably expressed on CHO- DHFR- cells. Conclusion: A large amount of GPI modified Met-RANTES fusion protein was expressed on CHO cells. GPI anchored form of Met-RANTES may be used as novel immunosuppressant for suppressing reaction in graft rejection.

The phenomenon of recessive absence was initiated by many malpractices in current medical educational system.In this article we try to analyze this phenomenon,speculate the subjective and objective reasons,propose countermeasures such as improving teachers quality,reforming traditional teaching mode,strengthening medical ethics education,enriching teaching means and addressing student-centered lectures.

Objective:To study the morphologic and phenotypic characters of a macrophage-tumor vaccine,and to observe the effect of macrophage-tumor vaccine on inducing CTL respose.Methods:The super-structure and the expression of CD14,CD68,CD80,CD86,MHC Ⅱ molecules of macrophage-tumor cells were detected with electron microscope,immunofluorescence staining and flow cytometry respectively.Meanwhile,H22 tumor cells were transplanted to the mice that had been immunized with different tumor vaccines.The weight and volume of tumors,the tumor cell injure rate and the level of LDH in culture supernatant were detected with direct measurement,MTT and selection methods.Results:The macrophage tumor vaccine cells were large cells with an irregular outline,and generally displayed pseudopodium,membrane folding,and vesicles on the cell surface.The predominant cytoplasmic organelles were lysosomes,secondary lysosomes and residual bodies.The percentage of CD14,CD68,CD80,CD86 and MHC Ⅱ positive cells within the differentiated population were 53.90%,98.60%,26.50%,90.20% and 25.40% respectively.The results of experiment in vivo revealed that the tumor forming rate,volume and weight of the group immunized with macrophage-tumor vaccine were much lower than that of control group and the group that were immunized with the macrophages that were induced by liquid paraffin (P0.05),the tumor weight and volume of the group immunized with the macrophage-tumor vaccine were lower than those of the group immunized with inactivated tumor cells(P

Objective To investigate the bid gene expression and cell death in brain after cerebral hypoxia-ischemia in neonatal rats and the effects of dexamethasone(DEX)on bid gene expression,so as to elucidate the possible mechanism of the neuroprotective effect of DEX pretreatment on rats following cerebral hypoxia-ischemia.Methods Twenty-four SD neonatal rats were divided randomly into hypoxia-ischemia brain damage(HIBD),normal,dexamethasone-pretreated and 9 g/L NaCl(NS)control group.The animal models of HIBD were made.Total RNA from ipsilateral cerebral hemisphere was extracted.Reverse transcription polymerase chain reaction(RT-PCR)was used to evaluate the level of bid gene expression after hypoxia-ischemia.Cerebral apoptosis was determined by terminal-deoxynucleotidy transferase mediated d-UTP nick end labeling(TUNEL).Results The levels of bid mRNA were higher in HIBD rats than those in normal rats.The number of positive apoptosis cells significantly increased in HIBD group(P

Objective To explore the cellular inhibitor of apoptosis protein 1(cIAP1)gene expression and Caspase-3 activity in brain after cerebral hypoxia-ischemia in neonatal rats and the influence of dexamethasone(DEX)on cIAP1 gene expression and Caspase-3 activity,so as to elucidate the possible mechanism of the neuro-protective effect of DEX pretreatment on rats following cerebral hypoxia-ischemia.Methods Twenty-four SD neonatal rats were divided randomly into hypoxic-ischemic brain damage group(HIBD group),normal group(NS group),dexamethasone-pretreated group(DEX group)and 9 g/L NaCl control group(NS group).The animal models of HIBD were made.Total RNA from ipsilateral cerebral hemisphere was extracted.Reverse transcription polymerase chain reaction(RT-PCR)was used to evaluate the level of cIAP1 gene expression after hypoxia-ischemia.Caspase-3 relative activity of brain tissue was determined by colorimetric assay.Results The levels of cIAP1 mRNA were lower in HIBD group than those in NS group.Caspase-3 relative activity significantly increased in HIBD group(P

Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

Objective The objective was to discussing the difference between the Pemberton osteotomy and Salter osteot?omy which performed in patients between the ages of 2 and 3 years who suffered from developmental dislocation of the hip (DDH). Methods A retrospective review of the results of operation treatment for DDH in 59 children (84 hips) from January 1998 to De?cember 2008 was conducted. There are 10 boys (14 hips) and 49 girls (70 hips). The age of the patients was between 2-3 years old at the time of treatment 2.5±0.4 years. Surgery consist of open reduction of the hip, capsulorraphy, shortening and derotational oste?otomy of proximal femur, and innominate osteotomy which include Pemberton osteotomy (33 hips), Salter osteotomy (51 hips). McKay and Severin modified criteria were used to assess the function and radiographic results of the hip. Results The average follow?up time was 5.6±3.5 years ranging from 2 to 16 years. According to Severin criteria at final follow?up, 78 hips (93%) had ex?cellent and good results;4 hips were fair and 2 hips poor result. The radiology results in Salter osteotomy were better than Pember?ton osteotomy (rate of excellent and good results 100%vs. 82%,χ2=7.43, P=0.003). According to the McKay criteria Salter osteoto?my and Pemberton osteotomy have no significant difference in latest follow?up (the satisfactory rate 100%vs. 97%,χ2=1.56, P=0.39). 18 hips (21%) had proximal femoral growth disturbance which 10 hips in Pemberton group, 8 hips in Salter group. There is no significant difference (χ2=2.54, P=0.17). Conclusion Open reduction, innominate osteotomy and proximal femoral osteotomy were effective procedures for the treatment of DDH in children between 2-3 years old. More attention should be taken in Pember?ton osteotomy to prevent the acetabular bony edge absorption.

AlM: To observe the effect of dacryocystorhinostomy for the treatment of chronic dacryocystitis using nasal endoscope and discuss the operation technique. METHODS: A retrospective clinical analysis was performed on the clinical data followed up for 6 ~12mo from 140 patients (169 eyes) with dacryocystorhinostomy for the treatment of chronic dacryocystitis using nasal endoscope. The effect of the treatment was evaluated and the operation technique for the treatment of chronic dacryocystitis using nasal endoscope was discussed.RESULTS: ln all of cases 155 eyes ( 91. 7%) were recovery, 3 eyes ( 1. 8%) were improved, and 11 eyes (6. 5%) were failure. The total efficiency was 93. 5%, there was no significant difference compared with traditional dacryocystorhinostomy group (χ2=3. 743, P>0. 05). CONCLUSlON: Dacryocystorhinostomy using nasal endoscope for treatment of chronic dacryocystitis has a good curative effect. Techniques including lacrimal sac location and size, colostomy position and size, treatment of colostomy mucosal flap and nasal disease, postoperative follow - up and physical condition of patients are likely to affect the operation curative effect.

Objective To investigate the effect of chest physiotherapy (CPT) on patients undergoing mechanical ventilation (MV).Methods A prospective randomized controlled trial (RCT) was conducted. Sixty-eight adult patients undergoing invasive MV over 48 hours admitted to intensive care unit (ICU) of Affiliated Hospital of Zunyi Medical College from December 2014 to October 2016 were enrolled, and they were divided into CPT group (n = 37) and control group (n = 31) by random number table. The patients in control group received routine physical therapy; while those in the CPT group received comprehensive CPT including manual lung inflation, vibration expectoration and early functional exercise etc. on the basis of the treatment in control group. Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and oxygenation index (PaO2/FiO2) before and after the treatment in both two groups were observed as well as the respiratory function and vital signs before and after CPT. The laboratory indicators after treatment, incidence of complications, duration of MV and the length of ICU stay in the two groups were recorded.Results The incidence of ventilator associated pneumonia (VAP) in the CPT group was significantly lower than that of control group (5.4% vs. 25.8%,P < 0.05), the patients in control group also had atelectasis, deep vein thrombosis and other complications, while no such complications were found in the CPT group. The duration of MV (hours: 77.4±41.0 vs. 133.9±117.2) and the length of ICU stay (hours: 134.4±71.4 vs. 207.4±177.7) in CPT group were significantly shorter than those of the control group (bothP < 0.05). There was no significant difference in APACHE Ⅱ score and PaO2/FiO2 before treatment between the two groups. After treatment for 2 days, the APACHE Ⅱ score in both groups was gradually decreased, and that in CPT group was more significantly, it was significantly lower than that of control group after treatment for 4 days (8.6±3.9 vs. 12.5±5.3,P < 0.05). The PaO2/FiO2 in the two groups was gradually increased after treatment. PaO2/FiO2 in CPT group was significantly increased at 3 days after treatment as compared with that before treatment [mmHg (1 mmHg = 0.133 kPa): 278.1±79.0 vs. 224.2±98.9], while PaO2/FiO2 in the control group did not appear significantly increased until after 4-day treatment (mmHg: 302.3±93.1 vs. 232.3±116.7, both P < 0.05). There was no significant difference in vital signs andrespiratory function parameters including tidal volume (VT), respiratory rate (RR), peak airway pressure (Ppeak) and mean airway pressure (Pmean) before and after treatment in CPT group excepting pulse oxygen saturation (SpO2) was significantly higher than that before treatment (0.985±0.016 vs. 0.978±0.018,P < 0.05), indicating that CPT treatment did not cause fluctuations in respiratory function and vital signs. Blood lactate in CPT group was significantly lower than that of control group (mmol/L: 1.10±0.79 vs. 1.32±1.09, P < 0.05), indicating that CPT treatment, especially early functional exercise, could improve the oxygen supply and limb circulation.Conclusion CPT treatment has some effect on prevention of VAP and other complications in patients undergoing MV, which could shorten the duration of MV and the length of ICU stay, and promote the recovery of patients.

Objective: To study the quality and transdermal properties of matrine microemulsion-based hydrogel (MBH) to provide basis for the development of the preparation.Methods: The stability of MBH was observed at 4 ℃ for 3 months and the changes of particle appearance, viscosity, pH and matrine content were observed.The transdermal permeation of MBH was investigated by a dual chamber permeation and diffusion device with excised mouse skin as the barrier.Taking rabbits as the experimental subjects, the irritation of MBH to the normal skin and damaged skin was investigated.Results: The appearance, viscosity, pH and matrine content of MBH at 4 ℃ in 3 months did not change significantly.In vitro transdermal test showed that MBH had a good penetration rate on mouse skin, and no skin irritation occurred after single or multiple administrations.Conclusion: MBH has good stability and high rate of transdermal penetration without skin irritation, which is a promising drug delivery system of matrine with good application prospects.