Objective:To investigate the influence of combining therapy of massage on sacro-2 to sacro-4 spinal cord reflexion area and anal dilatation therapy on constipation after cerebral apoplexy and promote the rehabilitation of defecation function in patients with constipation after stroke.Methods:Aandomized controlled study method was chosen, 80 patients with constipation after cerebral apoplexy treated in Guangzhou Traditional Chinese Medicine Hospital from August 2018 to July 2019 were selected as research subjects by convenience sampling. The enrolled patients were divided into 2 groups, 40 to the observation group and 40 to the control group by random number table method. On the basis of routine constipation related nursing measures, the control group was given anal dilatation therapy, while the observation group′s treatment was given massage on sacro-2 to sacro-4 spinal cord reflexion area combined with anal dilatation therapy. Before intervention and after intervention, the changes of therapeutic effect of constipation effect, constipation symptoms scores and the degree of neurological deficit were all compared between the two groups.Results:There were 22 males and 18 females in the observation group; age ranged from 40 to 75 (63.33 ± 8.49) years old; in the control group, there were 23 males and 17 females, aged 41-74 (63.03 ± 7.60) years old. Before intervention, there was no statistically significant difference in constipation symptom scores and neurological deficits between the two groups (both P>0.05). After intervention, the score of constipation symptoms was (3.80 ± 2.05) points and the score of the degree of neurological deficit was (7.58 ± 2.69) points in the observation group, both lower than (5.18 ± 3.52) points and (8.90 ± 2.73) points in the control group with statistically significant differences between the two groups ( t values were respectively -2.13 and -2.19, both P<0.05). The total efficacy of the observation group was 97.50% (39/40), which was significantly higher than 87.50% (35/40) of the control group with statistically significant differences between the two groups ( χ2 = - 2.11, P<0.05). Conclusions:Nursing intervention of massage on sacro-2 to sacro-4 spinal cord reflexion area combined with anus dilatation therapy on constipation after cerebral apoplexy can effectively reduce the symptoms of constipation and lowered the degree of neurological deficit, which is beneficial to the functional rehabilitation of patients with cerebral apoplexy.

With various diseases ravaging internationally, the demands for recombinant adenoviral vector (Adv) vaccines have increased dramatically. To meet the demand for Adv vaccine, development of a new cell culture process is an effective strategy. Applying hyperosmotic stress in cells before virus infection could increase the yield of Adv in batch culture mode. Emerging perfusion culture can significantly increase the yield of Adv as well. Therefore, combining the hyperosmotic stress process with perfusion culture is expected to improve the yield of Adv at high cell density. In this study, a shake flask combined with a semi-perfusion culture was used as a scaled-down model for bioreactor perfusion culture. Media with osmotic pressure ranging from 300 to 405 mOsm were used to study the effect of hyperosmotic stress on cell growth and Adv production. The results showed that using a perfusion culture process with a hyperosmotic pressure medium (370 mOsm) during the cell growth phase and an isosmotic pressure medium (300 mOsm) during the virus production phase effectively increased the yield of Adv. This might be due to the increased expression of HSP70 protein during the late phases of virus replication. The Adv titer in a bioreactor with such a process reached 3.2×10¹⁰ IFU/mL, three times higher than that of the traditional perfusion culture process. More importantly, this is the first time that a strategy of combining the hyperosmotic stress process with perfusion culture is applied to the production of Adv in HEK 293 cells. It also reveals the reason why the hyperosmotic stress process increased the yield of Adv, which may facilitate the process optimization of for producing other Adv in HEK 293 cells.
Humans ; HEK293 Cells ; Genetic Vectors/genetics* ; Batch Cell Culture Techniques ; Bioreactors ; Perfusion

Objective: To discuss the efficacy of computer aided design (CAD) and computer aided manufacturing (CAM) in the treatment of microgenia by osseous genioplasty. Methods: From July 2015 to January 2018, 17 cases (Male, n=5, Female, n=12), aged from 17 to 32 years, with the meanage of 24 years, were enrolled. All patients had microgenia, and underwent osseous genioplasty. Computed tomography (CT) was performed preoperatively and 7-10 days postoperatively for all cases. Based on preoperative CT data, CAD was applied to make individualized surgical plan, surgery simulation, todetermine the movement of mentum. CAM was used to prefabricate the individualized surgical templates and transfer the CAD into intraoperative guidance. The registration of CAD and postoperative CT were performed to evaluate the surgical accuracy. Results: All patients were satisfied with the surgical outcomes. The deformity of microgenia was significantly corrected. The Pg error was (0.24±0.08) mm, and the Me error was (0.15±0.17) mm. Conclusions The application of CAD increases the accuracy of surgical planning, and CAM provides areliable guidance with individualized template. Anoptimal surgical outcome can be achieved.

We constructed bicistronic expression system containing AH6 promoter, 5' UTR and its fore 38 bp sequence from Corynebacterium glutamicum, followed by a conserved Shine-Dalgarno (SD) sequence for xylanase expression. The two major secretory pathways signal peptide in C. glutamicum, Tat (CgR0949) and Sec (CspB) dependent signal peptide were added before xylanase for its secretion. Fed-batch cultivation was done in a 5 L jar for high-level xylanase secretion. The enzyme properties of the purified xylanase were then studied, including the effect of temperature and pH on its activity. The xylanase could be secreted into the culture supernatant when the Sec-dependent signal peptide CspB was used, but none was detected when CgR0949 was used. The secretory production level of xylanase in a flask was 486.2 U/mL and become 1 648.7 U/mL when in a 5 L jar, which was 3.4 fold as in the flask. The optimal pH and temperature of xylanase were pH 4.5 and 45 ℃, respectively. Its activity was 80% of initial activity after pretreatment at 4 ℃ for 24 h at pH 4-11, 95% after incubation below 50 ℃ for 15 min, and 20% when the temperature above 60 ℃. The xylanase could be efficiently secreted into the culture medium by C. glutamicum using its own genetic elements, and the secretion level could be improved through large-scale fed-batch cultivation. This bicistronic expression system can provide a useful tool for heterologous proteins secretion in C. glutamicum. In addition, the catalyze activity of xylanase could be further improved by enzyme properties study.
Corynebacterium glutamicum ; Promoter Regions, Genetic ; Protein Sorting Signals ; Protein Transport

We constructed different N-terminal truncated variants based on Bacillus acidopullulyticus pullulanase 3D structure (PDB code 2WAN), and studied the effects of truncated mutation on soluble expression, enzymatic properties, and application in saccharification. Upon expression, the variants of X45 domain deletion existed as inclusion bodies, whereas deletion of CBM41 domain had an effective effect on soluble expression level. The variants that lack of CBM41 (M1), lack of X25 (M3), and lack both of CBM41 and X25 (M5) had the same optimal pH (5.0) and optimal temperature (60 degrees C) with the wild-type pullulanase (WT). The K(m) of M1 and M5 were 1.42 mg/mL and 1.85 mg/mL, respectively, 2.4- and 3.1-fold higher than that of the WT. k(cat)/K(m) value of M5 was 40% lower than that of the WT. Substrate specificity results show that the enzymes exhibited greater activity with the low-molecular-weight dextrin than with high-molecular-weight soluble starch. When pullulanases were added to the saccharification reaction system, the dextrose equivalent of the WT, M1, M3, and M5 were 93.6%, 94.7%, 94.5%, and93.1%, respectively. These results indicate that the deletion of CBM41 domain and/or X25 domain did not affect the practical application in starch saccharification process. Furthermore, low-molecular-weight variants facilitate the heterologous expression. Truncated variants may be more suitable for industrial production than the WT.
Bacillus ; enzymology ; Glycoside Hydrolases ; metabolism ; Molecular Weight ; Protein Conformation ; Sequence Deletion ; Substrate Specificity ; Temperature

Objective To explore the effects of multiple geriatric syndromes on quality of life in the hospitalized elderly type 2 diabetes. Methods A cross sectional study was carried out in 397 elderly patients with type 2 diabetes mellitus by convenience sampling method. Each subject was assessed for general condition, quality of life as well as chronic pain, chronic constipation, urinary incontinence, high risk of falling, malnutrition, sleep disorder, polypharmacy. The influencing factors of quality of life were analyzed by multiple stepwise regression analysis. Results The patients averagely had 3.23 ± 1.51 geriatric syndromes, and 87.2% (346/397) of them had two or more geriatric syndromes. The average physical component summary of patients was (277.11±64.30) points, and mental component summary was (307.00 ± 60.46) points. The influencing factors of physical component of quality of life were quantity of geriatric syndromes, number of complications and course of disease, while the influencing factors of mental component of quality of life were quantity of geriatric syndromes, number of complications. Conclusions Multiple geriatric syndromes are closely related to the patients′quality of life to a greater degree as demographic factors and condition of diabetes. Nursing staffs should pay attention to the assessment and intervention of multiple geriatric syndromes in elderly patients with type 2 diabetes, so as to improve patients′quality of life effectively.

Cardiac Catheterization ; Ductus Arteriosus, Patent ; therapy ; Heart Septal Defects, Atrial ; therapy ; Humans ; Printing, Three-Dimensional ; Vena Cava, Inferior

Objective To explore the prognosis and management of atrial fibrillation (AF) in patients with atrial septal defect(ASD) accompanied by AF after transcatheter closure of ASD. Methods During the period from July 2010 to May 2013, a total of 24 patients with ASD accompanied by AF were admitted to authors’ hospital to receive transcatheter closure of ASD. Electrocardiogram (ECG), chest X-ray film and transthoracic echocardiography (TTE) were performed before and one day after the operation. Follow-up information was obtained through telephone or at out-patient clinic interview. Results Successful occlusion of ASD was obtained in all patients, and in no patient the AF rhythm turned to sinus rhythm after the procedure. In one patient preoperative AF turned to postoperative atrial flutter, and AF recurred in one case who had received transcatheter ablation of AF before the procedure. One female patient developed gastric bleeding during the course of orally taking warfarin, and she died of cerebral infarction at three days after ceasing the use of warfarin. Of the 24 patients, no anticoagulant drug was used in 5 (20.8%), oral administration of aspirin was given in 7 (29.2%), and oral medication of warfarin was employed only in 11 (45.8%). Conclusion The spontaneous conversion rate of AF is very low in patients with ASD complicated by AF after transcatheter closure of ASD. Postoperative medication of anticoagulation should be strictly standardized and carefully managed.

Objective To explore the application value of 64-slice spiral CT cardiac enhancement scanning in the morphological as-sessment of left atrial appendage(LAA).Methods Patients were selected retrospectively between April 16,2013 and May 1,2013, including 142 patients with sinus rhythms underwent MDCT coronary angiography,25 patients with atrial fibrillation underwent MDCT pulmonary vein-left atrial angiography.All images were post-processed,datas were measured and the morphology of left at-rial appendage were classified into different types.Results 167 patients were enrolled,including 88 males and 79 femals,age 53.7± 8.9 years.Anatomical relationship of the orifice of left atrial appendage to left superior pulmonary vein(LSPV)were classified as high type (superior to LSPV,2.4%),middle type(parallel to LSPV,86.2%),low type(inferior to LSPV,11.4%).LAA orifice could be classified into three types including oval(162,97%),round-like(3,1.8%)and drop-like(2,1.2%).LAA morphology could be classified in four types including Cauliflower(35,21%),Windsock(41,24.6%),Cactus(7,4.2%),Chicken-wing(84, 50.3%).Patients with atrial fibrillation have a larger LAA volume than who with sinus rhythm.Conclusion 64-slice spiral CT car-diac enhancement scanning can provide a vivid and intuitive reference to the anatomy of LAA and its adjacent structures,and it can also provide quantitative evaluation of LAA anatomical parameters,so it may have a nice significance in the direction of LAA occlu-sion.

ObjectiveTo classify the type of lumbosacral plexus nerve root injury.MethodsFrom November 2004 to August 2011,36 patients suffered with lumbarsacral plexus nerve root injury underwent surgical exploration in our department.There were 24 males and 12 females,aged from 7 to 49 years(average,29.5 years).By inductively analyzing the location and amount of nerve root injury,preoperative clinical manifestations and results of physical examination,the clinical typing of lumbarsacral plexus nerve root injury was made.ResultsLumbosacral plexus nerve root injury was classified into 6 types:total lumbosacral plexus nerve root injury (4 cases),lumbar plexus and upper sacral plexus nerve root injury (6 cases),sacral plexus nerve root injury (9 cases),upper sacral plexus nerve root injury (11 cases),lower sacral plexus nerve root injury(4 cases) and lumbar plexus injury(2 cases).There were 19 patients with total lumbosacral plexus nerve root injury,lumbar plexus and upper sacral plexus nerve root injury or sacral plexus nerve root injury,among which 73.7％(14/19) nerve root injury located in the spinal canal and all of them were nerve root avulsion or rupture.There were 17 patients with upper sacral plexus nerve root injury,lower sacral plexus nerve root injury or lumbar plexus nerve root injury,among which 64.7％ (11/17) nerve root injury located in intro-pelvic or pelvic sacral foramina,and all of them were distraction injury.ConclusionThis clinical typing is useful for the accurate diagnosis of lumbosacral plexus nerve root injury.In addition,it is also beneficial for judging the location and characteristics of nerve root injury.