The 1,095 bp gene encoding peroxidase from Coprinus cinereus was synthesized and integrated into the genome of Pichia pastoris with a highly inducible alcohol oxidase. The recombinant CiP (rCiP) fused with the a-mating factor per-pro leader sequence derived from Saccharomyces cerevisiae was secreted into the culture medium and identified as the target protein by mass spectrometry, confirming that a C. cinereus peroxidase (CiP) was successfully expressed in P. pastoris. The endoplasmic reticulum oxidoreductase 1 (Ero1) and protein disulfide isomerase (PDI) were co-expressed with rCiP separately and simultaneously. Compared with the wild type, overexpression of PDI and Erol-PDI increaseed Cip activity in 2.43 and 2.6 fold and their activity reached 316 U/mL and 340 U/mL respectively. The strains co-expressed with Erol-PDI was used to high density fermentation, and their activity reached 3,379 U/mL, which was higher than previously reported of 1,200 U/mL.
Coprinus ; enzymology ; Culture Media ; Cytoplasm ; Fermentation ; Glycoproteins ; metabolism ; Mass Spectrometry ; Mating Factor ; Oxidoreductases Acting on Sulfur Group Donors ; metabolism ; Peptides ; Peroxidases ; biosynthesis ; Pichia ; metabolism ; Protein Disulfide-Isomerases ; metabolism ; Protein Folding ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae Proteins ; metabolism

Objective This study was conducted to describe job stressors of nurses employed in military hospitals and relationship between job stressors and demography characters.Method 296 employed nurses were investigated by nurse job stressor questionnaires.Result Professional and career issues were the most frequently encountered job stressor among employed nurses,the other stressors in descending order were workload and time pressure,patient care and interaction,resource and environmental problems,management issues.Job stressors of nurses could be influenced by their age,job title,et al.Conclusion The finding suggest that effective stress management strategies should be relieve job stress.

Objective To analyze the platelet activating factor acetylhydrolase ( PAF-AH) activity of a gene product encoded by LA2144 gene of Leptospira interrogans ( L. interrogans) , to investigate the ex-pression and secretion of LA2144 protein in various cell cultures and to further understand its function in in-ducing internal hemorrhage in an animal model. Methods The DNA sample containing LA2144 gene was extracted from L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai and used as the template for gene cloning by PCR. The LA2144 gene without the signal sequence coding region was amplified by PCR and inserted into a prokaryotic expression construct for the protein expression. The expressed recombinant protein, rLep-PAF-AH, was purified by Ni-NTA affinity chromatography. Spectrophotometry was used to measure the hydrolytic activity, hydrolytic efficiency, Km and Kcat values of the rLep-PAF-AH protein in hydrolyzing PAF substrate. Real-time fluorescent quantitative RT-PCR ( qRT-PCR) and Western blot assay were performed to measure the expression of LA2144 gene at mRNA and protein levels in human umbilical vein endothelial cells (HUVEC), human monocytes (THP-1) and murine macrophages (J774A. 1) with L. interrogans strain Lai infection, respectively. Each syrian hamster was intravenously injected with 100 μg of LPS-free rLep-PAF-AH for two times. Hemorrhage in the lungs, livers and kidneys were observed in three days after the injection. Results The constructed prokaryotic expression system for LA2144 gene of L. inter-rogans strain Lai could highly express the rLep-PAF-AH upon the induction of IPTG. The purified rLep-PAF-AH showed high purity with a single protein band in gel as indicated by SDS-PAGE. The efficiency of 5 μg of rLep-PAF-AH in hydrolyzing PAF substrate was 26. 6 U/L with a Km value of 82. 79 μmol/L and a Kcat value of 0. 24 S-1 . The expression of Lep-PAF-AH at mRNA level in HUVEC, THP-1 and J774A. 1 cells were significantly elevated after co-culture with L. interrogans strain Lai for 1 or 2 hours (P<0. 05). A large amount of Lep-PAF-AH were detected in the supernatants from co-cultures of L. interrogans strain Lai with the three cell lines, but not from the culture of the spirochete in EMJH medium. The signs of hemor-rhage were observed in the lung of hamsters injected with rLep-PAF-AH, but not in tissue samples from liver and kidney. Conclusion The LA2144 gene product was characterized by a stronger PAF-AH activity. The expression of LA2144 gene at mRNA and protein levels in various cell lines were enhanced during L. interro-gans infection. Moreover, the rLep-PAF-AH could induce the pulmonary hemorrhage in hamsters. This stud-y indicated that the protein encoded by LA2144 gene was an important virulence factor causing hemorrhage in hosts during L. interrogans infection.

OBJECTIVE: To explore the risk factors of acute respiratory distress syndrome (ARDS) in patients with sepsis and to construct a risk nomogram model. METHODS: The clinical data of 234 sepsis patients admitted to the intensive care unit (ICU) of Tianjin Hospital from January 2019 to May 2022 were retrospectively analyzed. The patients were divided into non-ARDS group (156 cases) and ARDS group (78 cases) according to the presence or absence of ARDS. The gender, age, hypertension, diabetes, coronary heart disease, smoking history, history of alcoholism, temperature, respiratory rate (RR), mean arterial pressure (MAP), pulmonary infection, white blood cell count (WBC), hemoglobin (Hb), platelet count (PLT), prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB), D-dimer, oxygenation index (PaO₂/FiO₂), lactic acid (Lac), procalcitonin (PCT), brain natriuretic peptide (BNP), albumin (ALB), blood urea nitrogen (BUN), serum creatinine (SCr), acute physiology and chronic health evaluation II (APACHE II), sequential organ failure assessment (SOFA) were compared between the two groups. Univariate and multivariate Logistic regression were used to analyze the risk factors of sepsis related ARDS. Based on the screened independent risk factors, a nomogram prediction model was constructed, and Bootstrap method was used for internal verification. The receiver operator characteristic curve (ROC curve) was drawn, and the area under the ROC curve (AUC) was calculated to verify the prediction and accuracy of the model. RESULTS: There were no significant differences in gender, age, hypertension, diabetes, coronary heart disease, smoking history, alcoholism history, temperature, WBC, Hb, PLT, PT, APTT, FIB, PCT, BNP and SCr between the two groups. There were significant differences in RR, MAP, pulmonary infection, D-dimer, PaO₂/FiO₂, Lac, ALB, BUN, APACHE II score and SOFA score (all P < 0.05). Multivariate Logistic regression analysis showed that increased RR, low MAP, pulmonary infection, high Lac and high APACHE II score were independent risk factors for sepsis related ARDS [RR: odds ratio (OR) = 1.167, 95% confidence interval (95%CI) was 1.019-1.336; MAP: OR = 0.962, 95%CI was 0.932-0.994; pulmonary infection: OR = 0.428, 95%CI was 0.189-0.966; Lac: OR = 1.684, 95%CI was 1.036-2.735; APACHE II score: OR = 1.577, 95%CI was 1.202-2.067; all P < 0.05]. Based on the above independent risk factors, a risk nomograph model was established to predict sepsis related ARDS (accuracy was 81.62%, sensitivity was 66.67%, specificity was 89.10%). The predicted values were basically consistent with the measured values, and the AUC was 0.866 (95%CI was 0.819-0.914). CONCLUSIONS Increased RR, low MAP, pulmonary infection, high Lac and high APACHE II score are independent risk factors for sepsis related ARDS. Establishment of a risk nomograph model based on these factors may guide to predict the risk of ARDS in sepsis patients.
Humans ; Retrospective Studies ; Alcoholism ; Prognosis ; Respiratory Distress Syndrome ; Pneumonia ; Sepsis ; Intensive Care Units ; Procalcitonin ; Fibrinogen ; ROC Curve

This article describes the concept, theoretical basis, commonly used evaluation indicators of the intimacy enhancement therapy, as well as the application status and effects of the therapy among different cancer patients and their spouses, so as to provide a reference for the development of intimacy enhancement therapy suitable for Chinese culture and improve the quality of family life of cancer patients.

Major infectious disease epidemic continues to pose a threat to human health and society, and the effective establishment and implementation of an early warning system plays a key role in addressing public health security risks. At present, the research on early warning of infectious diseases in the academic community mainly focuses on early warning information system, early warning mechanism, laws and regulations of early warning of infectious diseases, and some studies lack specific suggestions on operation methods. By collating and summarizing the literature from 2002 to 2022, regarding the early warning system and mechanism of major infectious diseases, this paper focused on analyzing the public health ethical dilemmas existing in the early warning process and discussing how to strengthen the construction of the early warning system of infectious diseases, so as to lay the foundation for creating more scientific early warning schemes of infectious diseases.

The use of digital means has made the public health emergency management more efficient and convenient. However, in the practice of managing public health emergencies, there are dilemmas in the protection of personal health information, such as the imperfect legal system, the weakened right of informed consent and control, the lack of reasonable norms in the collection and use of information, and the disclosure of personal health information. To solve the dilemma of personal health information protection, it is necessary to improve the corresponding legal mechanism, strengthen the classification of health information, standardize the behavior of health information collection and use, enhance the technical support of personal health information protection, build a system combining law and technology, and protect the security of personal health information.

To enhance the production of glucose oxidase by recombinant Pichia pastoris, two strategies were developed, which were namely co-feeding of methanol and sorbitol and co-expressing of the protein disulfide isomerase (PDI) and Vitreoscialla hemoglobin (VHb). The volumetric activity reached 456 U/mL by using the strain X33/pPIC9k-GOD, in 5 liter fermentator, with the co-feeding of methanol and sorbitol, it was 0.2 fold higher than that only feeding by methanol. The improved strain was obtained by co-expressing PDI-VHb with GOD. While fermented in a 5 liter fermentator by feeding methanol and sorbitol, the activity of the improved strain reached 716 U/mL with a yield of 7 400 mg/L total soluble protein concentration. These results indicated that heterologous protein expression level can be enhanced by optimizing fermentation condition and co-expression molecular chaperon in Pichia pastoris.
Bioreactors ; Fermentation ; Glucose Oxidase ; biosynthesis ; Methanol ; Pichia ; metabolism ; Recombinant Proteins ; biosynthesis ; Sorbitol

Soluble cello-oligosaccharide with 2-6 oligosaccharide units is a kind of oligosaccharide with various biological functions, which can promote the proliferation of intestinal probiotics such as Bifidobacteria and Lactobacillus paracei. Therefore, it has a regulatory effect on human intestinal microbiota. In this study, a Cc 01 strain was constructed by expressing cellodextrin phosphorylase (CDP) in Escherichia coli. By combining with a previously constructed COS 01 strain, a three-enzyme cascade reaction system based on strains COS 01 and Cc 01 was developed, which can convert glucose and sucrose into cello-oligosaccharide. After optimization, the final titer of soluble cello-oligosaccharides with 2-6 oligosaccharide units reached 97 g/L, with a purity of about 97%. It contained cellobiose (16.8 wt%), cellotriose (49.8 wt%), cellotetrose (16.4 wt%), cellopentaose (11.5 wt%) and cellohexose (5.5 wt%). When using inulin, xylo-oligosaccharide and fructooligosaccharide as the control substrate, the biomass (OD₆₀₀) of Lactobacillus casei (WSH 004), Lactobacillus paracei (WSH 005) and Lactobacillus acidophilus (WSH 006) on cello-oligosaccharides was about 2 folds higher than that of the control. This study demonstrated the efficient synthesis of cello-oligosaccharides by a three-enzyme cascade reaction and demonstrated that the synthesized cello-oligosaccharides was capable of promoting intestinal microbial proliferation.
Humans ; Oligosaccharides ; Biomass ; Escherichia coli/genetics* ; Gastrointestinal Microbiome ; Glucose

OBJECTIVE To systematically evaluate the real-world effectiveness and safety of belimumab in the treatment of lupus nephritis （LN） in Chinese adult patients. METHODS Retrieved from PubMed， Embase， Web of Science， Cochrane Library， Wanfang data， CNKI， VIP and CBM， real-world studies on belimumab in the treatment of LN in Chinese adult patients were collected from the inception to July 7th， 2023. Two reviewers independently screened the literature， extracted data， and assessed the quality of the included studies. Meta-analysis was then performed using RevMan 5.3 software. RESULTS A total of 10 real- world studies were included， involving 253 Chinese adult patients with LN. The results of the meta-analysis demonstrated that the complete renal response rate， partial renal response rate， and the incidence of adverse reaction rate in Chinese adult patients with LN treated with belimumab were 61% （95%CI was 46%-76%， P＜0.000 01）， 23%（95%CI was 2%-44%， P＝0.03）， and 30% （95%CI was 16%-43%， P＜0.000 01）， respectively. Belimumab could reduce the 24-hour urinary protein （MD＝－1.71， 95%CI was －3.02-－0.40， P＝0.01）， urine protein-creatinine ratio （MD＝－1.76，95%CI was －2.06-－1.46，P＜0.000 01）， the systemic lupus erythematosus disease activity index （MD＝－8.63， 95%CI was －12.12-－5.13， P＜0.000 01）， and glucocorticoids dosage （MD＝－18.65， 95%CI was －31.82-－5.48， P＝0.006）. In addition， it could elevate the levels of complement C3 （MD＝0.19， 95%CI was 0.08-0.30， P＝0.000 6） and complement C4 （MD＝0.06， 95%CI was 0.02-0.09， P＝0.001）. However， belimumab could not improve the levels of serum creatinine and estimated glomerular filtration rate （P＞0.05）. CONCLUSIONS Belimumab has good efficacy and safety in Chinese adult patients with LN.