Objective:To compare the surgical efficacy of pericardial soft ring tricuspid valvuloplasty with DeVega and artificial valvuloplasty.Methods:227 patients undergoing tricuspid valvuloplasty due to rheumatic heart disease complicated with functional tricuspid valve insufficiency were retrospectively analyzed and divided into 3 groups according to tricuspid valvuloplasty dynamic cohort(pericardial ring group, 89 cases; the artificial flap ring group, 61 cases, and the DeVega group, 77 cases) were matched 1∶1 for propensity score(match A: pericardial ring group and artificial flap ring group; match B: pericardial ring group and DeVega group), the successful matching was included in follow-up and data collection, and cases with incomplete case data during follow-up were removed from the study cohort in pairs according to matching conditions. The results of follow-up 1 month, 6 months and 24 months after surgery were compared.Results:1 month after operation: the tricuspid valve regurgitant in all groups was significantly reduced or even disappeared compared with that before operation, and the right atrium and right ventricle were also smaller than that before operation, with statistical significance( P<0.05). 6 months after surgery: There was no statistical significance in the area of tricuspid regurgitation and right atrial/indoor diameter between all groups compared with the results one month after surgery( P>0.05), and there was no statistical significance in the recurrence rate of tricuspid regurgitation between all groups( P>0.05). 24 months after surgery: There were no significant differences in the recurrence rate of tricuspid regurgitation, area of tricuspid regurgitation and right atrial/indoor diameter between the two groups in matching A( P>0.05). There was no statistical significance in the right atrial/indoor diameter between the matched pericardial ring group and the Devega group, but the tricuspid valve regurgentation area of the Devega group at 24 months after surgery was higher than that of the Devega group at 1 month after surgery, and the difference was statistically significant( P<0.05). The regurgitation area and recurrence rate of tricuspid valve were significantly higher than those of pericardium-TVP group( P<0.05). Conclusion:Pericardial soft ring tricuspid valvuloplasty can effectively correct functional tricuspid valvuloplasty and reverse right heart remodeling, which is an effective tricuspid valvuloplasty.

Based on the demand of enterprise talents and the characteristics of manufacturing process management in biotechnology, in order to make the students acquire the ability to solve complex engineering problems in the production process, we developed a "Comprehensive Biotechnology Experiment" course, where two-step enzymatic production of l-aspartate and l-alanine were the key processes. In this course, we drew lessons from the site management of the production enterprise, performed the experimental operation mode of four shifts and three operations. The content of this course includes principles, methods and experimental techniques of several core curricula and the site management mode of enterprises. As to the evaluation, the summary of the experimental staff's handover records and the content of teamwork were examined and scored. Through teaching practice and continuous improvement, we developed a complete experimental teaching process and assessment mechanism. Overall, the Comprehensive Biotechnology Experiment course achieved good teaching effect, which may serve as a reference to promote the development of experimental teaching of biotechnology.
Humans ; Biotechnology ; Curriculum ; Students

The amino acid sequence of ancestral enzymes from extinct organisms can be deduced through in silico approach termed ancestral sequence reconstruction (ASR). ASR usually has six steps, which are the collection of nucleic acid/amino acid sequences of modern enzymes, multiple sequence alignment, phylogenetic tree construction, computational deduction of ancestral enzyme sequence, gene cloning, and characterization of enzyme properties. This method is widely used to study the adaptation and evolution mechanism of molecules to the changing environmental conditions on planetary time scale. As enzymes play key roles in biocatalysis, this method has become a powerful method for studying the relationship among the sequence, structure, and function of enzymes. Notably, most of the ancestral enzymes show better temperature stability and mutation stability, making them ideal protein scaffolds for further directed evolution. This article summarizes the computer algorithms, applications, and commonly used computer software of ASR, and discusses the potential application in directed evolution of enzymes.
Amino Acid Sequence ; Evolution, Molecular ; Phylogeny ; Proteins/genetics* ; Sequence Alignment

Objective To understand the extensions of pharmaceutical care at medical institutions in China.Methods From June to October 2018, 196 cases were collected regarding " exploring new areas of pharmaceutical care" , which ranks one of the ten innovative medical services under China Healthcare Improvement Initiative(2018-2010). By means of statistics on the frequency of the subject words in the cases, the authors identified hotspots of these hospitals in this regard, and used descriptive analysis to study the data. Results The 196 cases covered 23 items of pharmaceutical care, including three traditional" guaranteed drug supply" services, i.e., drug supply, drug dispensing and drug storage management; one traditional Chinese medicine supply service, namely decoction and delivery of traditional Chinese medicine decoction; 19 pharmaceutical professional technical services, namely, prescription(doctor′s advice)review, patient education and guidance, and prescription ( doctor′s advice) comment among others. Conclusions The connotation of pharmaceutical care is extending rapidly. Therefore we should enhance the professional value of pharmacists, establish training programs for pharmaceutical talents, establish incentive mechanism for pharmaceutical care, and promote the rapid development of patient-centered pharmaceutical care.

Objective To investigate the expressions of CD133 and p53 in colorectal cancer and their clin-ical significances. Methods The expressions of CD133 and p53 in 74 colorectal cancer patients were detected by the immunohistochemistry method. The relationships of CD133 and p53 with the clinicopathological parameters and prognosis were analyzed. Results The positive expression rates of CD133 and p53 in colorectal cancer tissues were 33.8%and 55.4%,respectively. The expression levels of CD133 and p53 were not related to age,sex,tumor location and histological type ,and but were significantly related to the histological differentiation ,TNM stage and distant metastasis(P<0.05,respectively). The Spearman correlation analysis showed that there was no correlation between the levels of CD133 and p53. Conclusions The high expressions of CD133 and p53 in colorectal cancer tissues were closely related to the histological differentiation and TNM stage. CD133 and p53 could be used as important biomarkers for the evaluation of malignant biological behavior,and the diagnosis and prognosis of colorectal cancer.

Objective To investigate the efficacy and tolerability of dose-dense chemotherapy with cisplatin plus 5-fluorouracil( PF regimen)in distant metastatic nasopharyngeal carcinoma( NPC)patients. Methods From April 1,2008 to April 30,2014,168 patients were assigned to traditional group(n = 83) and dose-dense group(n = 85)using digital random table in 1 : 1 ratio. All patients received PF regimen,and once every 28 days in traditional group and once every 14 days in dose-dense group. The primary endpoint was progression-free survival(PFS)and the secondary endpoint was overall survival(OS),toxicity and response rate. Results The median PFS,median OS,1,2,3-year survival rate,complete response rate and objective response rate were significantly improved in dose-dense group which were 13. 3 months,20. 2 months,80. 2% , 36. 0% ,16. 1% ,16. 5% ,84. 7% ,and those in control group were 10. 0 months,16. 1 months,59. 6% , 10. 1% ,0,3. 6% ,54. 2% respectively(χ2 = 24. 47,P = 0. 000;χ2 = 16. 65,P = 0. 000;χ2 = 8. 41,P =0. 004;χ2 = 16. 96,P = 0. 000;χ2 = 14. 91,P = 0. 000;χ2 = 7. 63,P = 0. 006;χ2 = 18. 47,P = 0. 000). The rates of grade 3-4 adverse events in dose-dense and traditional group were 38. 8% and 6. 0%(χ2 = 25. 81, P = 0. 000). Conclusion Dose-dense chemotherapy of PF is more efficient and acceptable toxic than the tra-ditional one. It can be a new treatment option for patients with distant metastases of NPC.

Escherichia coli AFP111 is a spontaneous mutant with mutations in the glucose specific phosphotransferase system (ptsG) in NZN111 (delta pflAB deltaldhA). In AFP111, conversion of xylose to succinic acid generates 1.67 molecule of ATP per xylose. However, the strain needs 2.67 molecule ATP for xylose metabolism. Therefore, AFP111 cannot use xylose due to insufficient ATP under anaerobic condition. Through an atmospheric and room temperature plasma (ARTP) jet, we got a mutant strain named DC111 that could use xylose under anaerobic condition in M9 medium to produce succinic acid. After 72 h, DC111 consumed 10.52 g/L xylose to produce 6.46 g/L succinic acid, and the yield was 0.78 mol/mol. Furthermore, the reaction catalyzed by the ATP-generating PEP-carboxykinase (PCK) was enhanced. The specific activity of PCK was 19.33-fold higher in DC111 than that in AFP111, which made the strain have enough ATP to converse xylose to succinic acid.
Atmosphere ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Industrial Microbiology ; Metabolic Engineering ; Mutation ; Plasma Gases ; pharmacology ; Succinic Acid ; metabolism ; Temperature ; Xylose ; metabolism

During the anaerobic fermentation by Escherichia coli AFP111 for succinic acid production, the viable cell concentration and productivity were decreased with the raising of succinic acid concentration. In order to restore cellular succinic acid productivity and prolong fermentation time, we collected strains and refreshed medium for repetitive succinic acid production. However, productivity is lower than that in the anaerobic fermentation before reusing strains. To enhance the productivity, strains were aerobically cultivated for 3 h in pure water before anaerobic fermentation. The activities of key enzymes were enhanced for better performance in producing succinic acid at anaerobic stage. After three rounds of repetitive fermentations, succinic acid concentration and yield reached to 56.50 g/L and 90% respectively. The succinic acid productivity was 0.81 g/(L x h), which was 13% higher than the repetitive fermentations without aerobic activation of the strains.
Aerobiosis ; Anaerobiosis ; Culture Media ; Escherichia coli ; genetics ; metabolism ; Fermentation ; Genetic Engineering ; Glucose ; metabolism ; Industrial Microbiology ; Succinic Acid ; metabolism

Escherichia coli BA002, in which the ldhA and pflB genes are deleted, cannot utilize glucose anaerobically due to the inability to regenerate NAD+. To restore glucose utilization, overexpression of nicotinic acid phosphoribosyltransferase (NAPRTase) encoded by the pncB gene, a rate-limiting enzyme of NAD(H) synthesis pathway, resulted in a significant increase in cell mass and succinate production under anaerobic conditions. However, a high concentration of pyruvate was accumulated. Thus, co-expression of NAPRTase and the heterologous pyruvate carboxylase (PYC) of Lactococcus lactis subsp. cremoris NZ9000 in recombinant E. coli BA016 was investigated. Results in 3 L fermentor showed that OD600 is 4.64 and BA016 consumed 35.00 g/L glucose and produced 25.09 g/L succinate after 112 h under anaerobic conditions. Overexpression of pncB and pyc in BA016, the accumulation of pyruvic acid was further decreased, and the formation of succinic acid was further increased.
Anaerobiosis ; Escherichia coli ; enzymology ; genetics ; metabolism ; Fermentation ; Genetic Engineering ; Glucose ; metabolism ; Industrial Microbiology ; Lactococcus lactis ; enzymology ; NAD ; metabolism ; Pentosyltransferases ; biosynthesis ; genetics ; Pyruvate Carboxylase ; biosynthesis ; genetics ; Succinic Acid ; metabolism

Sucrose phosphorylase (EC 2.4.1.7, Sucrose phosphorylase, SPase) can be produced by recombinant strain Escherichia coli Rosetta(DE3)/Pet-SPase. Crude enzyme was obtained from the cells by the high pressure disruption and centrifugation. Sucrose phosphorylase was purified by Ni-NTA affinity column chromatography and desalted by ultrafiltration. The specific enzyme activity was 1.1-fold higher than that of the crude enzyme, and recovery rate was 82.7%. The purified recombinant SPase had a band of 59 kDa on SDS-PAGE. Thermostability of the enzyme was shown at temperatures up to 37 degrees C, and pH stability between pH 6.0 and 6.7. The optimum temperature and pH were 37 degrees C and 6.7, respectively. The K(m) of SPase for sucrose was 7.3 mmol/L, and Vmax was 0.2 micromol/(min x mg). Besides, alpha-arbutin was synthesized from sucrose and hydroquinone by transglucosylation with recombinant SPase. The optimal conditions for synthesis of alpha-arbutin were 200 U/mL of recombinant SPase, 20% of sucrose, and 1.6% hydroquinone at pH 6-6.5 and 25 degrees C for 21 h. Under these conditions, alpha-arbutin was obtained with a 78.3% molar yield with respect to hydroquinone, and the concentration of alpha-arbutin was about 31 g/L.
Arbutin ; biosynthesis ; Catalysis ; Enzyme Stability ; Escherichia coli ; enzymology ; genetics ; Glucosyltransferases ; biosynthesis ; genetics ; metabolism ; Hydroquinones ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Sucrose ; metabolism