A new method of using photoactivable ester with azido group was described to immobilize urease on polyether sulfone(PES) film surface. The effects of photoactive enzyme concentration, temperature, pH, irradiation time on the activity of immobilized urease were investigated. Reused times and storage stability were also studied. The results showed that the surface concentration of urease immobilized on PES surface was about 0.33 mg/cm2. When the irradiation time was 5 minutes, the relative activity of immobilized urease was the highest and the activity increased with the increase of the concentration of photoactive urease solution. The optimum pH and temperature of immobilized urease were 7 and 50 degrees C respectively. The relative activity of immobilized urease was stable (50%) after 12 times reused at 50 degrees C.
Enzyme Stability ; Enzymes, Immobilized ; metabolism ; radiation effects ; Membranes, Artificial ; Photochemistry ; Polymers ; Sulfones ; Urease ; metabolism ; radiation effects

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.

In order to investigate the application of recombinant adeno-associated virus (rAAV) vector containing Tet regulation system and HSVtk gene in cancer gene therapy, pAAV/TRE/HSVtk/Tet-On was constructed and identified with PCR and restriction enzyme digestion. Packaging cells HEK293 were cotransfected with plasmids pAAV/TRE/HSVtk/Tet-On, pAAV-RC and pAAV-helper to produce infectious rAAV, and CsCl2 densitygradient centrifugation method was performed for purification and concentration of rAAV. The viruses were then transduced into MCF-7 cells. The results of dot blot hybridization indicate that the rAAV can transfer the target gene into MCF-7 cells. MTT assay showed that GCV could kill AAV-infected MCF-7 cells under the induction of Dox. The data demonstrated that rAAV containing Tet regulation system and HSVtk gene was successfully obtained, and could be used for further investigation of in vivo and in vitro experiments.
Cell Line, Tumor ; Dependovirus ; genetics ; metabolism ; Doxycycline ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Simplexvirus ; enzymology ; genetics ; Thymidine Kinase ; genetics ; Transfection

Objective To investigate the needs and its satisfaction degrees of the family members of ICU patients at the different stages. Methods 27 patients' family members who were complied to the selected standard were investigated by the Critical Care Family Needs Inventory (CCFNI) and the Needs Met Inventory (MNI) within 24 hours and after 72 hours separately after the patients were admitted in the ICU. Results The subscale of assurance of the patients was the important one of patients' family members' needs, so there was no statistical significance between the condition within 24 h and the condition after 72 h (P > 0.05). The conditions of support acquisition and comfort after 72 h were obviously higher than the conditions within 24 h, so the difference had statistical meaning (P < 0. 01). The degrees of satisfaction after 72 h was higher than that within 24 h. Sufficiency in information after 72 h was obviously higher than that within 24 h, showing a statisticeal meaning (P <0.01). Conclusions The needs and its satisfaction degrees of ICU patients' family members at the different stages are different. The ICU medical staff should pay full attention to all kinds of patients' needs, thereby increasing their satisfaction degrees.

OBJECTIVETo compare the effects and pharmacoeconomics of single-dose of ceftriaxone versus 3-day cefuroxime prophylaxis in patients undergoing gastric or colorectal resection.

METHODSThree hundred and five consecutive patients with gastric or colorectal cancer from 5 medical centers were randomly divided into ceftriaxone group (n = 153, receiving intravenously 1 g ceftriaxone 0.5 - 1 h prior to operation only) and cefuroxime group (n = 152, receiving 0.75 g cefuroxime preoperatively and the same dose q8h for 3 d). The patients' intra- and postoperative status, adverse responses and infectious complications were observed and documented, and pharmacoeconomic parameters were analyzed.

RESULTSThe disease distribution, operative procedures and patients' conditions in the 2 groups were comparable. No adverse responses to the test antibiotics were observed. Postoperative infectious complications occurred in 7 cases in the ceftriaxone group (4.58%) and 14 cases in the cefuroxime group (9.21%), respectively (P = 0.992), among which, 12 cases were surgical site infections (incisional, intra-abdominal): 2 cases in the ceftriaxone group (1.31%), and 10 cases in the cefuroxime group (6.58%), (chi(2) = 5.607, P = 0.018). The direct cost related to prevention and treatment of surgical site infections was 283.5 RMB in the ceftriaxone group and 811.1 RMB in the cefuroxime group (Z = 14.51, P = 0.000).

CONCLUSIONBoth ceftriaxone and cefuroxime are safe and effective for prevention of surgical site infections. Single-dose ceftriaxone prophylaxis is sufficient for gastric and colorectal operations, with a better cost-effectiveness ratio.

Adolescent ; Adult ; Aged ; Anti-Bacterial Agents ; administration & dosage ; economics ; therapeutic use ; Antibiotic Prophylaxis ; economics ; Ceftriaxone ; administration & dosage ; economics ; therapeutic use ; Cefuroxime ; administration & dosage ; economics ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Surgical Wound Infection ; prevention & control ; Treatment Outcome ; Young Adult

Objective To explore the application value of multimodal image fusion technology in the diagnosis and treatment of intrahepatic cholangiocarcinoma (ICC).Methods The retrospective descriptive study was conducted.The clinicopathological data of 11 patients with ICC who were admitted to Zhujiang Hospital of Southern Medical University between January and September 2018 were collected.There were 5 males and 6 females,aged (55 ± 12)years,with a range from 30 to 74 years.The data of contrast-enhanced computed tomography (CT) and magnetic resonance imaging (MRI) with gadoxetate disodium (Gd-EOB-DTPA) of the upper abdomen were respectively collected,and three-dimensional(3D) model of liver was constructed based on CT-MRI fusion images.The preoperative evaluation and surgical planning were carried out based on the different modal imaging examination technologies.The indocyanine green (ICG) molecular fluorescence imaging system and augmented reality navigation system were used to guide hepatectomy.Observation indicators:(1) preoperative evaluation;(2) intraoperative situations;(3) comparison between surgical planning based on the different model imaging technologies and actual surgical method;(4) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect postoperative complications up to November 2018.Measurement data with normal distribution were represented as Mean ± SD,measurement data with skewed distribution were represented as M (range).Count data were described as absolute number or percentage and comparisons were analyzed using the paired chi-square test.Results (1) Preoperative evaluation:the proportions of grade 3 and above branch vessels of the portal vein and hepatic vein system and tumor margin by enhanced CT examination and enhanced MRI examination were respectively 11/11,4/11 and 5/11,11/11,with statistically significant differences in above indicators (x2 =4.16,5.14,P＜0.05).The enhanced CT showed 11 liver cancer lesions and enhanced MRI showed 13 lesions (including 2 lesions not demonstrated by enhanced CT,with a maximum diameter ≤ 10 mm).The 3D model of liver based on CT-MRI fusion image:location,number,infiltrating range (tumor boundary),intrahepatic vascular distribution,variation and its spatial relationship with lesions could be stereoscopically,intuitively and comprehensively displayed.(2) Intraoperative situations:of 11 patients,11 lesions were explored with naked eyes;13 lesions were detected by ICG molecular fluorescence imaging system,including 2 lesions showing low uptake lesions in liver and gallbladder specific period by preoperative MRI examinations and intrahepatic metastasis cancer by pathologic examination.Of 11 patients,6 had naked-eye ischemia boundaries around related vessels of hepatic portal ligation;10 with anatomical hepatectomy had hepatic segments or hemihepatic boundary by ICG molecular fluorescence imaging system,including 2 using positive staining and 8 using anti-staining.Among 11 patients,3 (1 combined with local resection of hepatic segment Ⅷ metastases),2,2,1,1,1 and 1 underwent respectively left hepatectomy,left lateral lobectomy of liver,right hepatectomy,extended right hepatectomy,right lobectomy of liver,resection of partial hepatic segment Ⅷ and mesohepatectomy.Seven of 11 patients received regional lymph node dissection and 4 received simple lymph node biopsy.Of 11 patients,1 diagnosed as with bile leakage of liver section underwent suture and ligation treatment with 4-0 Prolene;10 didn't occurred bile leakage.The surgical margin of 11 patients was negative.The operation time,volume of intraoperative blood loss and duration of hospital stay were (240± 118)minutes,(275±249)mL and (13 ± 8) days,respectively.There was no blood transfusion in the perioperative period.(3) Comparison between surgical planning based on the different model imaging technologies and actual surgical method:surgical planning of 3D model based on CT,MRI and CT-MRI fusion image in 6,9 and 11 patients was respectively consistent with actual surgical method.(4) Follow-up:11 patients were followed up for 2-10 months,with a median time of 6 months.Three patients had postoperative complications,2 of which were found in Clavien-Dindo Ⅰ and Ⅱ,including 1 with pleural effusion + peritoneal effusion and 1 with pleural effusion,they were improved after conservative treatment;1 with complication of Clavien-Dindo Ⅲ (postoperative intra-abdominal bleeding) was improved by selective arterial embolization using percutaneous femoral artery puncture.There was no postoperative bile leakage,hepatic failure and death.Conclusion Multimodal image fusion technique is helpful to optimize the preoperative surgical planning,which can assist the recognition of important vessels and real-time navigation of hepatectomy during operation,and improve the safety of operation.

OBJECTIVE: To determine the effects of naoyian (NYA) serum on the expression of vascular endothelial growth factor (VEGF) protein in cultured rat cerebral microvascular endothelial cell (RCMEC) with hypoxia. METHODS: NYA serum was separated from rat heart which had been filled stomach with NYA successively for 3 days. The rat cerebral microvascular endothelial cells were taken from the Sprageu-Dawley rat brain at postborn 7 days. The rat cerebral microvascular endothelial cells were incubated at anaerobic incubator to establish the hypoxia models. The vigo of RCMEC was determined by MTT. The level of expression of VEGF protein was measured by cell immunohistochemistry and Western blot. RESULTS: The OD value of NYA serum group was higher than the control groups after hypoxia for 18 hours. VEGF protein was increased by hypoxia in cerebral microvascular endothelial cells (P < 0.05). The content of VEGF protein in NYA serum containing medium was more significantly elevated than those cultured in other control media (P < 0.01). CONCLUSION VEGF protein was induced by hypoxia in rat cerebral microvascular endothelial cells, and NYA could upregulate the expression of VEGF protein, which may be one of the protection mechanisms for cerebral microvascular endothelial cells.
Animals ; Animals, Newborn ; Capillaries ; cytology ; Cell Hypoxia ; Cells, Cultured ; Cerebral Cortex ; blood supply ; Drugs, Chinese Herbal ; pharmacology ; Endothelium, Vascular ; cytology ; metabolism ; Female ; Male ; Rats ; Rats, Sprague-Dawley ; Serum ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

Objective To develop a minimally invasive operating technique for the treatment of hypertensive thalamic hemorrhage. Methods The clinical data of 15 patients with hypertensive thalamic hemorrhage performed neuroendoscope-assisted micro-invasive surgical treatment in our hospitals from July 2007 to June 2010 were retrospectively analyzed; their treatment efficacy were also concluded. Results The mean operation time of these patients was (1.5±0.4) h and the amount of blood loss was 30-40 mL; the mean clearance rate of hematoma in the thalamus was (86.2 ±7.9)percentage. Patients were followed up and evaluated by Glasgow outcome scale for at least 3 months.Three patients (21.4%) showed good recovery, 4 (28.6%) moderate disability, 4(28.6%) severe disability and 2 (14.3%) vegetative survival; 1 patient (7.1%) died. Conclusion Neuroendoscope-assisted micro-invasive surgical treatment is a fast and minimally invasive operating technique with little blood loss in the treatment of hypertensive thalamic hemorrhage.

Objective:To explore the association between triglyceride-glucose (TyG) index and carotid artery plaque in patients with type 2 diabetic kidney disease (DKD).Methods:Clinical data of 620 DKD patients admitted in the Department of Endocrinology, the Affiliated Huai′an First People′s Hospital of Nanjing Medical University from August 2018 to August 2022 were retrospectively analyzed, including 366 cases with carotid artery plaque and 254 cases without carotid plaque. According to TyG index quartile patients were divided into Q 1,Q 2, Q 3 and Q 4 groups with TyG index<8.94,≥8.94 and<9.44,≥9.44 and<9.96, and≥9.96, respectively. The prevalence of carotid plaque in DKD patients with different TyG index levels was analyzed. The relationship between TyG index and carotid plaque occurrence in DKD patients were analyzed with Logistic regression analysis and restricted cubic lines (RCS). Results:The age, course of disease, smoking rate, SBP, HbA1c, TG, BUN, eGFR and TyG indexes in carotid plaque group were significantly higher than those in non-carotid plaque group (all P<0.05). Binary logistic regression analysis showed that age, disease course, smoking rate, SBP, HbA1c, TG, BUN, low eGFR and TyG index were independent influencing factors for carotid plaque ( OR=1.05, 1.05, 1.88, 1.01, 1.09, 1.11, 1.09, 0.99 and 1.28, all P<0.05). The risk of carotid plaque in DKD patients in Q 3 and Q 4 groups was 2.20 and 2.50 times higher than that in Q 1 group. After adjusting for age, sex, course of disease, smoking, BMI, blood pressure (SBP and DBP), blood lipids (TC, HDL and LDL) and renal function, the risk of carotid plaque in DKD patients in Q 3 and Q 4 groups was higher than that in Q 1 group ( OR=1.95 and 2.24). RCS analysis showed that the correlation between TyG index and the risk of carotid plaque in DKD patients was linear(χ 2=0.40, P=0.527), and DKD patients with TyG index>9.95 had a higher risk of carotid plaque. Conclusions:TyG index is significantly elevated in DKD patients with carotid plaque, and TyG index is an independent risk factor for the occurrence of carotid plaques in DKD patients.

Although radiation therapy is a cornerstone of modern management of malignancies, various side effects are inevitably linked to abdominal and pelvic cancer after radiotherapy. Radiation-mediated gastrointestinal (GI) toxicity impairs the life quality of cancer survivors and even shortens their lifespan. Hydrogen has been shown to protect against tissue injuries caused by oxidative stress and excessive inflammation, but its effect on radiation-induced intestinal injury was previously unknown. In the present study, we found that oral gavage with hydrogen-water increased the survival rate and body weight of mice exposed to total abdominal irradiation (TAI); oral gavage with hydrogen-water was also associated with an improvement in GI tract function and the epithelial integrity of the small intestine. Mechanistically, microarray analysis revealed that hydrogen-water administration upregulated miR-1968-5p levels, thus resulting in parallel downregulation of MyD88 expression in the small intestine after TAI exposure. Additionally, high-throughput sequencing showed that hydrogen-water oral gavage resulted in retention of the TAI-shifted intestinal bacterial composition in mice. Collectively, our findings suggested that hydrogen-water might be used as a potential therapeutic to alleviate intestinal injury induced by radiotherapy for abdominal and pelvic cancer in preclinical settings.
Animals ; Body Weight ; Down-Regulation ; Gastrointestinal Microbiome ; Gastrointestinal Tract ; Humans ; Hydrogen ; Inflammation ; Intestine, Small ; Mice ; Microarray Analysis ; Oxidative Stress ; Pelvic Neoplasms ; Quality of Life ; Radiotherapy ; Survival Rate ; Survivors