Thermal Shift Assays was applied for high-throughput screening optimized buffer system used in the gel filtration chromatography for purification of recombinant TBEV serine protease. The optimization included kinds of salt and pH. It was optimized new buffer remarkably improved the quality of the purified TBEV serine protease by increasing its monomer from 75% to 99%. This result may improve future crystal studies of this protein.

Purpose The glucosamine and chondroitin compound tablets were prepared. The anti-inflammatory and analgesic effects of glucosamine and chondrotin compound tablets were investigated. Methods The proportion of glucosamine and chondroitin was 5:4 to prepare compound tablets. The anti-inflammatory effect was investigated with carrageen-induced rats paw edema and cotton granuloma in rats. The analgesic effect was investigated using the pain models of mice which were induced by 0.6% acetic acid. Results Compared with control group the degree of paw edema in the low, middle and high dose group was decreased ( P < 0.05). Fourtreatment groups compared with control group respectively at the weight of granuloma were also markedly reduced ( P < 0.05) and the writhing number of mice induced by acetic acid was decreased ( P < 0.05 ) . Conclusion glucosamine and chondroitin compound tablets have anti-inflammatory effect and analgesic effect on models induced by acetic acid.

Objective To investigate the self-management behaviors in patients with liver cirrhosis and analyze its influencing factors. Methods Totally 103 liver cirrhosis patients were recruited by convenience sampling method. They were investigated by using Self- management Questionnaire for Chinese patients with liver cirrhosis. Results The total score of self-management behaviors was 63.69 ± 13.60. All subscales scoring from high to low in turn were medication, diet management, condition monitoring and daily management. Conclusions The self-management level in patients with liver cirrhosis of decompensation period need to be improved, nursing staff should pay attention to effect evaluation during the hospital health education, to ensure that patients can master disease self-management knowledge, to improve the self-management behavior of patients with liver cirrhosis.

Background Conventional chemical method to fix the entire eyeball results in the edema of retinal nerve fiber layer and therefore influent the evaluation of tissue structure.Research showed that microwave irradiation can avoid this phenomenon. Objective This study was to investigate the appropriate energy and time duration of microwave irradiation for fixation of rabbit eyes specimen and compare the influence of different fixation methods on retinal structure. Methods Twenty-two New Zealand white rabbits were sacrificed by using venous air embolism.The eyeballs were enucleated from the rabbits and immersed in chemical reagents with glacial acetic acid,methyl aldehyde and chloroform for 2 days as the control group.The eyeballs were immersed in 400 ml physiological balance solution immediately,and then were fixed by microwave irradiation for 80,160,240 and 320 seconds under the lower power as the microwave irradiation groups.In the microwave irradiation+chemical fixation group,the specimens were immersed in chemical reagents for 1-2 hours after the microwave radiation.The morphology of retinal structure was examined and area of retinal detachment was calculated by hemotoxylin and eosin staining under the light microscope. Results A better fixation effect was obtained in the retinal section of microwave radiation for 240 seconds in comparison with that of the traditional chemical reagent fixation method for 2 days.The retina detachment ratio was 16.3%±11.5% for simple microwave method,50.0%±24.5% for chemical method,and 6.7%±7.8% for microwave+chemical method.showing a significant difference among 3 groups(F=32.43,P=0.000).Retinal staining was clear and retinal structure was almost normal in the specimens of microwave+chemical fixation group with 200-240 seeonds microwave irradiation and 1-2 hours chemical fixation. Conclusion Microwave irradiation method is a more ideal way for the fixation of retina because of taking short duration,lower toxicity and better staining.The combination of microwave radiation and chemical reagents can acquire an excellent quality of retinal section.

Chronic myeloid leukemia (CML) is characterized by the accumulation of active BCR-ABL protein.Imatinib is the first-line treatment of CML;however,many patients are resistant to this drug.In this study,we aimed to compare the differences in expression patterns and functions of time-series genes in imatinib-resistant CML cells under different drug treatments.GSE24946 was downloaded from the GEO database,which included 17 samples of K562-r cells with (n=12) or without drug administration (n=5).Three drug treatment groups were considered for this study:arsenic trioxide (ATO),AMN107,and ATO+AMN107.Each group had one sample at each time point (3,12,24,and 48 h).Time-series genes with a ratio of standard deviation/average (coefficient of variation) ＞0.15 were screened,and their expression patterns were revealed based on Short Time-series Expression Miner (STEM).Then,the functional enrichment analysis of time-series genes in each group was performed using DAVID,and the genes enriched in the top ten functional categories were extracted to detect their expression patterns.Different time-series genes were identified in the three groups,and most of them were enriched in the ribosome and oxidative phosphorylation pathways.Time-series genes in the three treatment groups had different expression patterns and functions.Time-series genes in the ATO group (e.g.CCNA2 and DAB2)were significantly associated with cell adhesion,those in the AMN107 group were related to cellular carbohydrate metabolic process,while those in the ATO+AMN107 group (e.g.AP2M1) were significantly related to cell proliferation and antigen processing.In imatinib-resistant CML cells,ATO could influence genes related to cell adhesion,AMN107 might affect genes involved in cellular carbohydrate metabolism,and the combination therapy might regulate genes involved in cell proliferation.

OBJECTIVETo investigate and clarify the key surgical techniques and evaluate clinical effectiveness of modular cementless femoral stems(LINK MP modular stem) in complex revision hip arthroplasty.

METHODSFrom January 2002 to December 2013, 470 patients in Chinese PLA General Hospital received revision hip arthroplasties using MP stems. Among them, 246 patients were followed for an average of 5.7 years (ranged, 0.5 to 11 years). There were 148 males and 98 females, with an average age of 62.5 years old (ranged from 26 to 83 years old). The most common reason for revision was aseptic loosening of femoral stems (107 cases, 43.5%), followed by prosthetic joint infection (104 cases, 42.3%). Other reasons included unexplained hip or thigh pain, periprosthetic femoral fracture, prosthesis wear and dislocation. According to Paprosky classification of femoral deficiency, there were 171 cases of type IIIA (69.5%), 14 cases of type I, 39 cases of type II, and 22 cases of type IIIB. The average preoperative Harris hip score (HHS) was 37.4 ± 7.5. In addition, radiographic presentation, degree of patients' subjective satisfaction, and clinical effectiveness were assessed as well.

RESULTSThe average Harris score was im- proved to 87.8 ± 4.2 after operation,with significant statistical difference when compared to preoperative score (t = 92.13,P = 0.00). Analysis of patients' subjective satisfaction for leg length discrepancy, stability and overall degree of satisfaction showed that the number of patients with great satisfaction were 27, 60, 61 respectively. The most common complication was the periprosthetic fracture (intraoperatively 21 cases and postoperatively 8 cases). Other complications included infection (7 cases), dislocation (5 cases), numbness possibly caused by incomplete sciatic nerve injury (6 cases) and thigh swelling (3 cases). The X-ray taken at the final follow-up revealed that prosthesis subsidence occurred in 4 cases, 2 of which needed a second revision.

CONCLUSIONThe application of modular cementless femoral stems (LINK MP modular stem) in complex revision hip arthroplasty (especially in cases with the serious defect of proximal femoral bone) shows good results at both the short-term and long-term follow-up, especially in the revision for prosthetic infection with the high success rate. In order to avoid complications and improve clinical outcomes,the surgeons should carefully select proper surgery candidates,perform the procedure with care, and master the key surgical techniques.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; methods ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Patient Satisfaction ; Postoperative Complications ; therapy

OBJECTIVETo investigate the effect of salidroside on hypoxia-induced apoptosis of corpus cavernosum smooth muscle cells (CCSMCs) in rats.

METHODSRat CCSMCs were cultured in vitro by the enzyme digestion method and identified by immunofluorescent staining of anti-alpha-SMA and anti-Desmin. The non-toxic dose of salidroside was determined by MTT assay. Low-oxygen mixed gas (1% O2, 5% CO2, and 94% N2) was piped into a modular incubator chamber to induce hypoxia. The CCSMCs were divided into a normal, a hypoxia, and a 32 microg/mL salidroside intervention group. The apoptosis of the CCSMCs was detected by flow cytometry and the expression of the caspase-3 protein determined by Western blot.

RESULTSThe majority of the CCSMCs were positive for alpha-SMA and Desmin at immunofluorescent staining. Salidroside at < 32 microg/ml produced no obvious toxicity to CCSMCs. Compared with the normal control group, the rates of early and late apoptosis of CCSMCs were both increased significantly in the hypoxia group ([12.77 +/-1.41]% vs [18.69 +/- 1.29]%, P < 0.01 and [14.63 +/- 2.00]% vs [21.03 +/- 1.530]% , P < 0.05). Western blot showed a markedly increased expression of cleaved caspase-3 (P < 0.01). Intervention with 32 microg/ml salidroside significantly reduced hypoxia-induced early apoptosis of CCSMCs ([13.46% +/- 1.87]%, P < 0.01) and decreased the expression of cleaved caspase-3 (P < 0.01).

CONCLUSIONSalidroside can reduce the expression of cleaved caspase-3 and inhibit hypoxia-induced apoptosis of CCSMCs in rats.

Animals ; Apoptosis ; drug effects ; physiology ; Caspase 3 ; metabolism ; Cell Hypoxia ; physiology ; Cells, Cultured ; Glucosides ; pharmacology ; Humans ; Male ; Myocytes, Smooth Muscle ; cytology ; drug effects ; enzymology ; Penis ; cytology ; drug effects ; Phenols ; pharmacology ; Rats

Objective To investigate the morbidity of anomalous coronary origin from the opposite coronary sinus, which may cause sudden death of young athletes in Chinese population.And to identify the imaging characteristics of this anomaly and its clinical significance combined with literature review.Methods The computed tomographic coronary angiography (CTCA) database at TEDA International Cardiovascular Hospital was reviewed.All of the patients diagnosed with isolated anomalous origin of a coronary artery from the opposite sinus of valsalva (anomalous origin of coronary artery, AOCA) and subsequent coursing between the pulmonary artery and the aorta were collected from 14 343 Chinese individuals.The location of anomalous coronary origin, the shape and course of the proximal ectopic arterial segments were identified.The nonatherosclerostic stenotic caliber of the segments and the angle between the ectopic coronary artery and the adjacent aortic wall were assessed.Results Seventy-four patients of AOCA (including the left or right single coronary artery) were diagnosed using CTCA.Among the 74 cases, the potentially serious course of the ectopic coronary artery between the pulmonary artery and the aorta were identified in 59 individuals.Fifty-six cases of ectopic right coronary with interarterial course (anomalous origin of right coronary artery, AORCA) and three patients with anomalous origin of the left coronary artery (AOLCA) were found, including two cases judged as potentially serious origin of either single left coronary artery (n=1) or single right coronary artery (n=1).The morbidity of the potentially serious anomalous origin of coronary artery in Chinese population was established as 4.1‰ (59/14 343).In the subgroup of AORCA, the lumen of initial ectopic segment was frequently compressed and stenotic.In 29 cases (52.7%) the stenosis of the lumen were more than 50%, and in 3 cases (5.4%) the stenosis of the ectopic coronary artery were more than 70%.The incidence of AORCA was 17.7 times higher (56/3) than that of AOLCA in Chinese population.Conclusions Nowadays CTCA is considered the most useful imaging technique in identifying the origin, shape and course of ectopic initial segment of the coronary artery.The significance of these anatomic characteristics, that may induce sudden death in Chinese young athletes, need to be investigated further.

Objective To investigate the anti-inflammatory and analgesic effects of combined application of glucosamine(GS)and chondroitin sulfate(CS).Methods The acute and chronic anti-inflammatory effects of combined application of GS and CS were observed respectively through carrageen-induced paw edema and cotton-induced inguinal granuloma of rats.The analgesic effect of combined application of GS and CS was investigated by the mouse pain model induced by 0.6% acetic acid Results As compared with the control group,the degree of paw edema and the weight of granuloma in combined application of GS and CS group were significantly reduced(P<0.05 and P<0.01,respectively);and the writhing number of mice decreased significantly(P<0.05).Conclusion Combined application of glucosamine and chondroitin sulfate demonstrates obviously anti-inflammatory and analgesic effects.

BACKGROUND: Imatinib has a significant pro-apoptosis effect on chronic myelogenous leukemia (CML), but there are still some patients being resistant to it. Human umbilical cord mesenchymal stem cells (hUC-MSCs) affect the apoptosis of a variety of hematologic malignancies. However, the impacts of hUC-MSCs on the apoptosis of CML cells induced by imatinib remain unclear.OBJECTIVE: To investigate whether hUC-MSCs have an influence on the apoptosis of K562 cells induced by imatinib and to reveal the possible underlying mechanism.METHODS: K562 cells were cultured with hUC-MSCs or/and imatinib. Cellular apoptosis was measured with Annexin-V and PI staining by flow cytometry analysis. The protein expressions of Bax, Bcl-2, caspase-3, caspase-9 and cleaved-PARP in K562 cells were detected by western blot assay. Pan-caspase inhibitor Z-VAD-FMK was used to block apoptosis in each group, and during this process the effect of caspase apoptosis signaling pathway was detected.RESULTS AND CONCLUSION: The apoptosis of K562 cells was enhanced, when imatinib was combined with hUC-MSCs. Western blot analysis showed that the expression of pro-apoptotic protein Bax was enhenced and the expression of anti-apoptotic protein Bcl-2 was suppressed. Furthermore, the cleaved forms of caspase-9, caspase-3 and PARP in K562 cell were higher in the hUC-MSCs+imatinib group than in the imatinib group. The apoptosis of K562 cells induced by the hUC-MSCs combined with imatinib was significantly inhibited by Z-VAD-FMK. In conclusion, these findings indicate that hUC-MSCs can enhance imatinib-induced apoptosis of K562 cells by activating caspase apoptosis signaling pathway.