Objective To investigate the enteral nutrition(EN)complications in ICU critically ill patients and their risk factors.Methods The relevant information of 376 EN patients from three tertiary hospitals in Qingdao from September 2015 to April 2016 were retrospectively analyzed.The incidences of EN complications and their risk factors were analyzed.Results In these 376 patients,the complications included diarrhea(n=117,31.12％),constipation(n=89,23.67％),and reflux(n=102,27.13％).The incidence of diarrhea showed significant difference in terms of the application of antibiotics(OR=3.55),age(OR=1.03),fasting(OR=4.07),oral potassium preparations(OR=2.91),nasal feeding rate(OR=1.03),and nasal feeding volume(OR=1.002).The incidence of constipation showed significant difference in terms of nasal fluid type(OR=7.79),analgesics(OR=10.34),age(OR=1.06),nasal feeding rate(OR=0.95),and nasal feeding volume(OR=0.97).The incidence of reflux showed significant difference in terms of the use of analgesics(OR=2.143),mechanical ventilation(OR=2.071),nasal feeding pathway(OR=1.838),age(OR=1.025),nasal feeding rate(OR=1.042),and nasal feeding volume(OR=1.001).ConclusionApplication of antibiotics,increased age,fasting,oral potassium preparations,increased nasal feeding rate,and increased daily feeding volume are risk factors of EN-related diarrhea;short peptide EN solution,application of analgesics,and increased age are risk factors of EN-related constipation for enteral nutrition patients;and mechanical ventilation,application of nasogastric tube,increased age,increased nasal feeding rate,and increased nasal feeding are risk factors of EN-related relux in enteral nutrition patients.

Objective To investigate the protective effects of lentivirus mediated Bcl-2-associated athanogene 1L (BAG-1L) over-expression on human neuroblastoma cells (SH-SYSY) induced by hypoxia/re-oxygenation,and to study its effect on the phosphoinositide 3 kinase serine/threonine protein kinase (PI3K/AKT) pathway.Methods SH-SYSY cells were cultured in vitro,and the cells at logarithmic phase were collected,and they were divided into recombined lentiviral infection group [infected by lentivirus containing BAG-1L and green fluorescent protein (GFP) gene],vector control group (infected by lentivirus containing GFP without BAG-1L gene) and cell control group (non-infection).Western Blot was used to detect the expression of BAG-1L in target cells after infection for 48 hours.SH-SY5Y cells were subjected to hypoxia for 8 hours and re-oxygenation for 24 hours,then the cell counting kit-8(CCK-8) was used to detect the cell activity,and the apoptosis was detected by flow cytometry after allophycocyanin labeled annexin V/7-amino actinomycin D (Annexin V-APC/7-AAD) staining.Western Blot was used to detect the protein expressions of BAG-1L,heat shock protein 70 (HSP70),AKT and phosphorylated AKT (p-AKT).Results After infection for 48 hours,exogenous BAG-1L protein bands were observed in recombined lentiviral infection group,but not observed in cell control group and vector control group.After hypoxia/re-oxygenation treatment,the cell viability in recombined lentiviral infection group was significantly higher than that in cell control group and vector control group (A value:0.689 ± 0.036 vs.0.425 ± 0.013,0.400 ± 0.012),apoptosis was significantly decreased [apoptosis rate:(26.97 ± 1.82)％ vs.(36.60± 1.45)％,(35.77 ± 3.74)％],the protein levels of BAG-1L,HSP70 and p-AKT were significantly increased [BAG-1L protein (gray value):2.405 ± 0.167 vs.0.529 ± 0.141,0.601 ± 0.099;HSP70protein (gray value):0.997±0.123 vs.0.634±0.091,0.584±0.106;p-AKT protein (gray value):1.234±0.118 vs.0.661 ± 0.210,0.712 ± 0.199,all P ＜ 0.01],but the protein level of AKT was slightly increased (gray value:1.103 ± 0.269vs.0.646 ± 0.188,0.791 ± 0.326) without statistically significant differences (both P ＞ 0.05).There was no significant difference in all parameters between cell control group and vector control group (all P ＞ 0.05).Conclusion Lentivirus mediated BAG-1L gene over-expression can protect nerve cells against hypoxic injury and apoptosis,and the protective effect may be related to the activation increase of pathway on PI3K/AKT.

Objective To evaluate the respiration-induced dosimetric variance in 3DCRT for midthoracic esophageal carcinoma, in order to guide the radiation oncologist to choose the expansion margin. Methods Ten patients with mid-thoracic esophageal carcinoma were scanned by multi-spiral CT simulator respectively in free breathing ( FB), breath-hold after normal inspiration and expiration ( IBH and EBH )with the same scanning range. Then the CT images of three series were transferred to the treatment planning system. The target volume was outlined following the same standard. Plan1 was designed in the images of FB and transported completely to the images of IBH and EBH as Plan2 and Plan3 respectively to observe the dosimetric variance in target volume. Results For GTV, there was a statistical difference only in V100 of the three plans ( H = 6.423, P = 0.040 ) and no significant difference was found in other indexes. For CTV, the V100 and V95 were better in Plan1 (F=3.992, P=0.030; H=9.920, P=0.007) and no significant difference was found in other indexes. While ()TV, the Dmin, V100 and V95 was better in Plan1 ( F = 3.677, P = 0.039; F = 4.539, P = 0.020; H = 6.846, P = 0.033 ) and no significant difference was found in other indexes. There were no significant differences in all the indexes for the spinal cord and lung in the three plans. Conclusions The change in dose distribution was not so much with the standard expansion. It can meet the needs of clinical treatment.

Objective To evaluate the respiration-induced target volume motion in 3D-CRT for mid-thoracic esophageal carcinoma in order to guide the radiation oncologist to choose the expansion marginfor ITV.Methods Ten patients with mid-thoracic esophageal carcinoma were scanned by multi-spiral CTsimulator respectively in free breathing(FB),breath.hold after normal inspiration and expiration(IBH and EBH)with the same scanning range.Then the CT images of three series were transfefred to the treatmentplanning system.The target volume was outlined following the same standard.The motion of the centerpoint of GTV,the center point of each slice of GTV and the edge of the GTV in selected slice weremeasured respectively to obtain the comprehensive value of GTV motion。in order to find the appropriate IMvalue according to the 95％confidence interval of the GTV motion.Results①The GTV motion betweenIBH and EBH was(0.19±0.16)cm in the left.right direction,(0.54±0.19)cm in the cranial andcaudal irection.and(0.16±0.14)cm in anterior.posterior directions for the center of GTV,.For thecenter point of each slice of GTV.they ere(0.19±0.15)cm,(0.54±0.16)cm,(0.16±0.13)cm in three directions above.respectively.For the edge of the GTV in selected slice.they were(0.26±0.19)cm,(0.54±0.18)cm,(0.24±0.19)cm,respectively.The comprehensive value of GTV motion between IBH and EBH was(0.23±0.17)cm,(0.54±0.17)cm,(0.21±0.17)cm.respectively.The 95％confidence interval was 0.21-0.25 cm.0.53-0.56 cm and 0.19-0.22 cm in three directions.②The direction of GTV motion：No motion was noticed in 8.2％.while 73.3％to the right side and 18.5％to the left side in the left-right direction when IBH were compared with EBH.100％were moved to caudal in the the cranial and caudal direction[(0.54±0.17)cm].In the anterior-posterior direction,no motion was noticed in 8.2％,while 16.6％to the posterior and 75.2％to the anterior when IBH were compared with EBH.③The GTV motion was correlated with the vafiance of 1ung volumes in IBH-EBH(r=0.683,P=0.032)and not with GTV volume and length.Conclusions Respiration can induce target volume motion in 3 DCRT for mid-thoracic esophageal carcinoma.Compared to EBH.the GTV tends to move to the caudal,the anterior and the ight side in IBH.

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.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on voltage-gated calcium channels on the cell membrane of pheochromocytoma cell 12 (PC 12 cells) induced by ischemic/hypoxia and its mechanisms.Methods PC12 cells at logarithmic phase were collected,which were challenged with hypoxia for 6 hours and reoxygenation for 12 hours to stimulate the nerve cells suffered ischemia/reperfusion pathological process in vitro.PC12 cells were divided into non-infection group,infected by lentivirus containing green fluorescent protein (GFP) without HSP70 gene lentivirus control group (GFP lentivirus control group),and infected by lentivirus containing HSP70 and GFP gene recombined lentiviral infection group (HSP70 recombined lentiviral infection group).Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot were used to determine mRNA and protein expressions of L-type voltage-gated Ca2+ channel subunits cav1.2 and cav1.3,receptor gated channel subunits NR1 and NR2,and Na+/Ca2+ exchange (NCX) in PC12 cells.Results After being challenged with hypoxia/reoxygenation,the mRNA and protein expressions of cav1.2,NR1 and NR2 in the PC12 cells were significantly lower in HSP70 recombined lentiviral infection group than those of GFP lentivirus control group and non-infection group [cav1.2 mRNA (2-△△Ct):3.13 ± 0.46 vs.5.12 ± 0.52,5.13 ± 0.66;NR1 mRNA (2-△△△Ct):1.61 ± 0.44 vs.3.23 ±0.82,3.31 ±0.78;NR2 mRNA (2-△△Ct):2.09±0.41 vs.3.91 ±0.64,3.88±0.62;cav1.2 protein (gray value):2.82±0.39 vs.3.98±0.23,3.96±0.24;NR1 protein (gray value):1.84±0.35 vs.2.79±0.21,2.86±0.23;NR2 protein (gray value):0.87±0.24 vs.1.57±0.31,1.33±0.44;all P ＜ 0.01].But there were no statistical differences in the mRNA and protein expressions of cav1.2,NR1 and NR2 between GFP lentivirus control group and non-infection group (all P ＞ 0.01).There were no statistical differences in the mRNA and protein expressions of cav1.3 and NCX among non-infection group,GFP lentivirus control group and HSP70 recombined lentiviral infection group [cav1.3 mRNA (2-△△Ct):4.82 ± 0.32,4.72 ± 0.36,4.82 ± 0.29;NCX mRNA (2-△△Ct):3.49 ± 0.78,3.47 ± 0.71,3.56 ± 0.65;cav 1.3 protein (gray value):2.63±0.40,2.64±0.39,2.68±0.39;NCX protein (gray value):3.27±0.48,3.34±0.48,3.31 ±0.42;all P ＞ 0.01].Conclusion Exogenous HSP70 affects the expression of L-type voltage-gated Ca2+ channel subunit cav1.2 and receptor gated channel subunits NR1 and NR2,which may protect PC12 cells from the injury caused by ischemic/hypoxia.

Objective To investigate the protective effects of Bcl-2 associated with athanogene-1 (BAG-1) gene on human neuroblastoma cells (SH-SY5Y) injury induced by hypoxia/reoxygenation,and its influence on heat shock protein 70 (HSP70) expression.Methods The SH-SYSY cells at logarithmic growth phase were collected.Lenti virus mediated RNA interference (RNAi) technology was used to suppress the BAG-1 expression.The cells screened out can be divided into four groups:the cell control group with no lentivirus infection,lentivirus control group (containing only fluorescein protein lentivirus infection),BAG-1 siRNA group (BAG-1 siRNA silencing group),including BAG-1 siRNA-α group and BAG-1 siRNA-β group with lentivirus containing fluorescein protein (GFP) but at different BAG-1 siRNA target sites of silencing.Western Blot was used to detect the protein expression of BAG-1 and HSP70 in target cells after infectious recombination lentivirus for 72 hours;the Cell Counting Kit-8 (CCK-8) was used to detect the activity of four different group cells after hypoxia;the flow cytometry was used to detect the cell apoptosis;the HSP70 mRNA transcription level were detected by real-time fluorescent quantitative reverse transcription-polymerase chain reaction (RT-qPCR) respectively in each group.Results After lentiviral infection for 72 hours,the Western Blot results showed that in the two BAG-1 siRNA silencing groups,the interference effect of BAG-1 siRNA-β group was superior to that of BAG-1 siRNA-α group.The cell viability of each group showed an increase followed by a decrease with the prolongation of hypoxia time,and reaching the peak at 8 hours.After hypoxia for 8 hours being given,the cell viability in BAG-1 siRNA-β group was significantly lower than that of the cell control group,lentivirus control group and BAG-1 siRNA-α group (A value:0.59 ±0.09 vs.0.94±0.12,0.90± 0.11,0.91± 0.14,P ＜ 0.01);the cell apoptosis rate was obviously higher in BAG-1 siRNA-β than that in the above three groups [(34.63 ± 3.46)％ vs.(14.83 ± 3.75)％,(19.93 ± 6.49)％,(16.40± 1.18)％,all P ＜ 0.01].There were no statistically significant differences in the HSP70 protein level and mRNA transcription level between BAG-1 siRNA-3 grroup,and the cell control group,lentivirus control group and BAG-1 siRNA-α group respectively (all P ＞ 0.05).Conclusion BAG-1 gene can play a role in protection of hypoxia nerve cells,reduce the apoptosis,and its protective effect can be independent of HSP70 gene.

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 investigate the long-term prognosis and failure mode of patients with esophageal squamous cell carcinoma who achieved clinical complete remission (cCR) after receiving radical radio (chemo) therapy.Methods:Clinical data of 183 patients with esophageal squamous cell carcinoma eligible for inclusion criteria who received treatment in our hospital from January 2009 to December 2012 were retrospectively analyzed. The factors that affected the long-term prognosis of patients were identified, and the failure mode of cCR patients and the prognosis after failure were analyzed. SPSS 19.0 statistical software was used for data analysis.Results:As of the follow-up date, the 1-, 3-, 5-, and 10-year overall survival (OS) rates and disease-free survival (DFS) rates of the entire group were 83.1%, 53.4%, 36.2%, 12.8% and 68.9%, 45.9%, 30.5%, 12.0%, respectively. The median OS and DFS were 41.3 months and 33.4 months. The results of multivariate analysis showed that cT staging, cN staging and prescribed dose were the independent factors affecting the OS ( P=0.001, <0.001, 0.003); hoarseness, lesion length, cT staging, cN staging and prescribed dose were the independent factors that affected the DFS ( P=0.002, 0.033, 0.009, <0.001, 0.003). In the whole group, 72 cases (39.3%) had local regional recurrence, 58 cases (31.7%) had distant metastasis, and 26 cases (14.2%) had local regional recurrence with distant metastasis. Among 104 patients after treatment failure, the prognosis of patients receiving salvage treatment was significantly better than that of their counterparts receiving maintenance treatment ( χ2=39.153, P<0.001). Conclusions:The long-term prognosis of patients with esophageal squamous cell carcinoma who achieved cCR after receiving radical radio (chemo) therapy is still unsatisfactory. Clinically, it is necessary to strengthen the clinical observation and follow-up of these patients. The main treatment failure mode of cCR patients is local regional recurrence. Active salvage treatment can significantly improve clinical prognosis of these patients.

Objective:To investigate the stability and feasibility of improved silent MRA technique based on hybrid-arterial spin labeling(ASL) for imaging intracranial arterial stenosis.Methods:From September 2019 to May 2020, totally 35 patients with suspected intracranial vascular stenosis in Department of Neurology of Northern Jiangsu People′s Hospital were enrolled in this study. Silent MRA and improved silent MRA based on hybrid-ASL technique were performed respectively. The acquisition noise (noise measurement and subjective score) of two kinds of MRA examination were evaluated respectively. Two neuroradiologists performed image quality scoring and signal-to-noise ratio (SNR) measurement of intracranial arteries (including internal carotid artery, vertebrobasilar artery, anterior cerebral artery, middle cerebral artery, and posterior cerebral artery) in the two kinds of MRA images using a double-blind, completely randomized method. Independent sample t-test was used to compare the image quality and SNR of two kinds of MRA images in each segment. Two experts assessed the degree of stenosis at the site of confirmed intracranial artery stenosis. Kappa test was used to assess interobserver and intermodel agreement. Results:There was no significant difference in acquisition noise between improved silent MRA and silent MRA ( P>0.05). In all five segments measured, the image quality scores of internal carotid artery [(4.40±0.49)scores], anterior cerebral artery[(4.30±0.33)scores] and middle cerebral artery [(4.46±0.34)scores] in improved silent MRA were higher than those in silent MRA images [(4.02±0.43)scores, (4.02±0.31)scores, (4.02±0.31)scores; t=2.825, 2.877, 1.683, all P<0.05)]. The SNR of internal carotid artery (9.11±1.23) and middle cerebral artery (8.77±1.87) in improved silent MRA images was higher than that in silent MRA images (7.83±1.33, 8.06±2.67, respectively; t=11.154, 3.268, both P<0.05). A total of 24 patients (38 lesions) with intracranial vascular stenosis were diagnosed by CTA. Improved silent MRA (Kappa=0.89, 95%CI 0.82-0.95) and silent MRA (Kappa=0.85, 95%CI 0.77-0.92) were highly consistent among observers in evaluating the degree of cerebrovascular stenosis.The results of improved silent MRA were highly consistent with those of CTA (Kappa=0.92, 95%CI 0.87-0.98), and those of silent MRA were highly consistent with those of CTA (Kappa=0.85, 95%CI 0.77-0.92). Conclusions:The improved silent MRA is feasible to improve the imaging quality and signal uniformity through efficient marking based on keeping the low noise features. In the diagnosis of intracranial stenosis and occlusive disease, the stability of improved silent MRA imaging improves the diagnostic efficiency of stenosis to a certain extent.