To screen and identify a chitosanase with high stability, we cloned the chitosanase gene from Bacillus atrophaeus with a high protease yield from the barren saline-alkali soil and expressed this gene in Escherichia coli. The expressed chitosanase of B. atrophaeus (BA-CSN) was purified by nickel-affinity column chromatography. The properties including optimal temperature, optimal pH, substrate specificity, and kinetic parameters of BA-CSN were characterized. The results showed that BA-CSN had the molecular weight of 31.13 kDa, the optimal temperature of 55 ℃, the optimal pH 5.5, and good stability at temperatures below 45 ℃ and pH 4.0-9.0. BA-CSN also had good stability within 4 h of pH 3.0 and 10.0, be activated by K⁺, Na⁺, Mn²⁺, Ca²⁺, Mg²⁺, and Co²⁺, (especially by Mn²⁺), and be inhibited by Fe³⁺, Cu²⁺, and Ag⁺. BA-CSN showcased the highest relative activity in the hydrolysis of colloidal chitosan, and it had good hydrolysis ability for colloidal chitin. Under the optimal catalytic conditions, BA-CSN demonstrated the Michaelis constant K_m and maximum reaction rate V_max of 9.94 mg/mL and 26.624 μmoL/(mL·min), respectively, for colloidal chitosan. In short, BA-CSN has strong tolerance to acids and alkali, possessing broad industrial application prospects.
Bacillus/genetics* ; Hydrogen-Ion Concentration ; Escherichia coli/metabolism* ; Glycoside Hydrolases/biosynthesis* ; Substrate Specificity ; Enzyme Stability ; Chitosan/metabolism* ; Temperature ; Kinetics ; Cloning, Molecular ; Bacterial Proteins/biosynthesis* ; Recombinant Proteins/genetics*

Implementation evaluation frameworks are essential tools in implementation science for assessing the quality and effectiveness of evidence-based interventions. This paper systematically reviews eight internationally representative evaluation frameworks, outlining their development backgrounds and structural features. It then compares their usability, applicability, and testability. Two case studies are presented to illustrate how evaluation frameworks can be integrated with process models and determinant frameworks to enhance the understanding and guidance of complex interventions. This paper aims to offer practical guidancefor selecting and applying evaluation frameworks, thereby supporting the advancement of implementation science in both local and global health contexts.

Objective:To explore the correlation between serum hepcidin antimicrobial peptide (HAMP), secreted phosphoprotein 1 (SPP1), and regulator of G protein signaling 2 (RGS2) levels and the clinical pathological characteristics of gastric cancer patients, and their predictive value for postoperative recurrence or metastasis.Methods:A total of 92 gastric cancer patients treated at Handan First Hospital from March 2021 to March 2023 were selected as the gastric cancer group, and 92 healthy individuals who underwent physical examinations during the same period were selected as the control group. The serum levels of HAMP, SPP1 and RGS2 were compared between the two groups. According to the mean levels of HAMP, SPP1, and RGS2 in the serum of gastric cancer patients, they were divided into HAMP high level group and HAMP low level group, SPP1 high level group and SPP1 low level group, RGS2 high level group and RGS2 low level group. The clinicopathological characteristics of gastric cancer patients with different levels of HAMP, SPP1 and RGS2 were compared respectively. After a median follow-up of 18 months, gastric cancer patients were divided into a non-recurrence or metastasis group ( n=59) and a recurrence and metastasis group ( n=33) based on whether the tumor recurred or metastasized. The serum levels of HAMP, SPP1, and RGS2 were compared between the two groups of patients. The predictive value of HAMP, SPP1 and RGS2 for postoperative recurrence or metastasis in patients with gastric cancer was analyzed by using the receiver operator characteristic (ROC) curve. Results:Compared with the control group, the gastric cancer group had higher levels of serum HAMP [ (52.28±5.44) ng/ml vs. (31.22±4.18) ng/ml] and SPP1 [ (55.96±6.43) ng/ml vs. (36.99±5.25) ng/ml] ( t=29.44, P<0.001; t=21.92, P<0.001), and lower level of RGS2 [ (3.72±0.66) mg/L vs. (5.11±0.87) mg/L) ] ( t=12.21, P<0.001). There were statistically significant differences in maximum tumor diameter ( χ2=13.07, P<0.001; χ2=6.71, P=0.010; χ2=10.56, P=0.001), TNM staging ( χ2=7.42, P=0.006; χ2=6.36, P=0.012; χ2=5.39, P=0.020), lymph node metastasis ( χ2=23.41, P<0.001; χ2=6.52, P=0.011; χ2=13.11, P<0.001), and differentiation degree ( χ2=9.01, P=0.003; χ2=7.97, P=0.005; χ2=15.29, P<0.001) between the gastric cancer patients in the HAMP high level group ( n=44) and the HAMP low level group ( n=48), the SPP1 high level group ( n=43) and the SPP1 low level group ( n=49), and the RGS2 high level group ( n=50) and the RGS2 low level group ( n=42). Compared with the non-recurrence or metastatic group, the recurrence and metastatic group had higher levels of serum HAMP [ (59.26±5.66) ng/ml vs. (48.37±4.28) ng/ml] and SPP1 [ (62.85±6.36) ng/ml vs. (52.11±5.38) ng/ml] level ( t=10.40, P<0.001; t=8.60, P<0.001), and lower level of RGS2 [ (3.01±0.48) mg/L vs. (4.12±0.69) mg/L] ( t=8.19, P<0.001). ROC curve analysis showed that the area under the curve (AUC) values of serum HAMP, SPP1, and RGS2 levels alone for predicting postoperative recurrence or metastasis in gastric cancer patients were 0.777, 0.813, and 0.778, respectively. The AUC value of the combination of the three indicators for predicting postoperative recurrence or metastasis in gastric cancer patients was 0.871. The predictive efficacy of the combination of the three indicators for predicting postoperative recurrence or metastasis in gastric cancer patients was better than that alone ( Z=2.51, P=0.035; Z=2.61, P=0.032; Z=2.71, P=0.029) . Conclusions:The levels of HAMP and SPP1 in the serum of gastric cancer patients increase, while the level of RGS2 decreases, and the levels of the three are related to the maximum tumor diameter, TNM staging, lymph node metastasis and differentiation degree, and their combined detection has higher predictive value for postoperative recurrence or metastasis in gastric cancer patients.

ObjectiveTo investigate the contamination status of Staphylococcus aureus in food and the presence of enterotoxin genes in Jiading District， Shanghai， and to provide a basis for the prevention and treatment of foodborne Staphylococcus aureus disease. MethodsFrom 2021 to 2023， 15 types of food were sampled for S. aureus testing， and the presence of five enterotoxin genes， including sea⁃see， was tested in the strains. ResultsOut of 705 food samples， 88 （12.48%） were positive for S. aureus. S. aureus was detected in 12 of the 15 food types， with the three food types with the highest positive rates being cold noodles （45.00%）， raw poultry （26.25%）， and vegetable salads （20.00%）. The enterotoxin gene carriage rate was 32.95% in food strains. The carriage rates for sea， seb， and sec were 7.95%， 12.50%， and 14.77%， respectively. Neither sed nor see was detected. The detection rate of strains carrying two types of enterotoxin genes was 2.27%. The enterotoxin carriage rates in strains from vegetables， beverages， and raw meat were 57.14%， 40.00%， and 30.00%， respectively. ConclusionThe S. aureus detection rate in food in Jiading District is much higher than the national average. The enterotoxin gene carriage rates are high， with food strains carrying sea， seb， and sec， with sec being the most prevalent. There is a need to enhance monitoring of S. aureus and enterotoxins， especially in high-risk foods such as noodles， vegetables， and non-packaged beverages.

Objective To build the early predictive model for chronic kidney disease(CKD)in hypertension and diabetes patients in the community.Methods The CKD patients were recruited from 4 health care centers in 4 urban areas in Kunming.The control group was residents without hypertension and diabetes(n = 1267).The disease group was residents with hypertension and/or diabetes(n = 566).The questionnaire survey,physical examination,laboratory testing,and 5 SNPs gene types in the PVT1 gene.The risk factors,which were filtered with logistics regression,were used to build predictive models.Four machine learning algorithms were built:support vector machine(SVM),random forest(RF),Na?ve Bayes(NB),and artificial neural network(ANN)models.Results Thirteen indicators included in the final diagnostic model:age,disease type,ethnicity,blood urea nitrogen,creatinine,eGFR from MDRD,ACR,eGFR from EPI2009,PAM13 score,sleep quality survey,staying-up late,PVT1 SNP rs11993333 and rs2720659.The accuracy,specificity,Kappa value,AUC of ROC,and PRC of ANN are greater than those of the other 3 models.The sensitivity of RF is the highest among 4 types of machine learning.Conclusions The ANN predictive model has a good ability of efficiency and classification to predict CKD with hypertension and/or diabetes patients in the community.

Objective:The status quo and related factors of human papillomavirus (HPV) infection in multiple parts of men who have sex with men (MSM) in Tianjin were analyzed.Methods:Using the cross-sectional survey method, a total of 600 MSM were recruited in Tianjin from September to December 2023, including 200 (18-26 years old), 300 (27-45 years old), and 100 (≥46 years old) stratificaed by age group. Questionnaire survey was used to collect knowledge and behavioral information related to demography and STD prevention and treatment. Exfoliated cells from genital, perianal and oral cavity were collected for HPV typing detection. Logistic regression model was used to analyze the related factors of HPV infection.Results:The infection rate of HPV was 53.67% (322/600) in MSM, with high-risk type as 41.17% (247/600), low-risk type as 27.17% (163/600). The infection rate of HPV in both multiple and single types was 26.83% (161/600).The infection rates of genotypes covered by nine-valent HPV vaccine, quadrivalent HPV vaccine and bivalent HPV vaccine were 36.83% (221/600), 25.17% (151/600) and 9.50% (57/600), respectively.The infection rates of genital HPV, high-risk or low-risk types were 24.67% (148/600), 16.67% (100/600) and 11.00% (66/600), respectively. The infection rates of perianal HPV, high-risk or low-risk types were 42.00% (252/600), 30.17% (181/600) and 20.50% (123/600), respectively. The infection rates of oral HPV high-risk and low-risk types were 3.17% (19/600), 2.33% (14/600) and 1.17% (7/600), respectively. Divorce or widowhood (a OR=2.38, 95% CI: 1.26-4.50), history of homosexual anal intercourse in the past 6 months (a OR=2.28, 95% CI: 1.00-5.50), and use of dependent drugs (including new psychoactive substances) (a OR=1.62, 95% CI: 1.02-2.58) were risk factors for genital HPV infection. College degree/university degree or above (a OR=0.36, 95% CI: 0.16-0.81) was a protective factor for perianal HPV infection, and a history of drug dependence (a OR=1.85, 95% CI: 1.19-2.88) was a risk factor. Conclusions:MSM in Tianjin was a high-risk group for HPV infection, and the HPV infection in the perianal area was more severe than in the genital area and oral area.Marital status, education level, history of homosexual anal sex in the past 6 months, and history of drug dependence were related factors of HPV infection among MSM.

Objective:To construct and validate a nomogram model for predicting the risk of 28-day mortality in sepsis patients.Methods:A retrospective cohort study was conducted. 281 sepsis patients admitted to the department of intensive care unit (ICU) of the 940th Hospital of the Joint Logistics Support Force of PLA from January 2017 to December 2022 were selected as the research subjects. The patients were divided into a training set (197 cases) and a validation set (84 cases) according to a 7∶3 ratio. The general information, clinical treatment measures and laboratory examination results within 24 hours after admission to ICU were collected. Patients were divided into survival group and death group based on 28-day outcomes. The differences in various data were compared between the two groups. The optimal predictive variables were selected using Lasso regression, and univariate and multivariate Logistic regression analyses were performed to identify factors influencing the mortality of sepsis patients and to establish a nomogram model. Receiver operator characteristic curve (ROC curve), calibration curve, decision curve analysis (DCA), and clinical impact curve (CIC) were used to evaluate the nomogram model.Results:Out of 281 cases of sepsis, 82 cases died with a mortality of 29.18%. The number of patients who died in the training and validation sets was 54 and 28, with a mortality of 27.41% and 33.33% respectively. Lasso regression, univariate and multivariate Logistic regression analysis screened for 5 independent predictors associated with 28-day mortality. There were use of vasoactive drugs [odds ratio ( OR) = 5.924, 95% confidence interval (95% CI) was 1.244-44.571, P = 0.043], acute physiology and chronic health evaluation Ⅱ (APACHEⅡ: OR = 1.051, 95% CI was 1.000-1.107, P = 0.050), combined with multiple organ dysfunction syndrome (MODS: OR = 17.298, 95% CI was 5.517-76.985, P < 0.001), neutrophil count (NEU: OR = 0.934, 95% CI was 0.879-0.988, P = 0.022) and oxygenation index (PaO 2/FiO 2: OR = 0.994, 95% CI was 0.988-0.998, P = 0.017). A nomogram model was constructed using the independent predictive factors mentioned above, ROC curve analysis showed that the AUC of the nomogram model was 0.899 (95% CI was 0.856-0.943) and 0.909 (95% CI was 0.845-0.972) for the training and validation sets respectively. The C-index was 0.900 and 0.920 for the training and validation sets respectively, with good discrimination. The Hosmer-Lemeshoe tests both showed P > 0.05, indicating good calibration. Both DCA and CIC plots demonstrate the model's good clinical utility. Conclusions:The use of vasoactive, APACHEⅡ score, comorbid MODS, NEU and PaO 2/FiO 2 are independent risk factors for 28-day mortality in patients with sepsis. The nomogram model based on these 5 indicators has a good predictive ability for the occurrence of mortality in sepsis patients.

OBJECTIVE: To explore the genetic basis for fetus with bilateral lateral ventriculomegaly. METHODS: Fetus umbilical cord blood and peripheral blood samples of its parents were collected. The fetus was subjected to chromosomal karyotyping, whilst the fetus and its parents were subjected to array comparative genomic hybridization (aCGH). The candidate copy number variation (CNV) were verified by qPCR, Application goldeneye DNA identification system was used to confirm the parental relationship. RESULTS: The fetus was found to have a normal karyotype. aCGH analysis indicated that it has carried a 1.16 Mb deletion at 17p13.3, which partially overlapped with the critical region of Miller-Dieker syndrome (MDS), in addition with a 1.33 Mb deletion at 17p12 region, which is associated with hereditary stress-susceptible peripheral neuropathy (HNPP). Its mother was also found to harbor the 1.33 Mb deletion at 17p12. qPCR analysis confirmed that the expression levels of genes from the 17p13.3 and 17p12 regions were about the half of that in the normal control, as well as the maternal peripheral blood sample. Parental relationship was confirmed between the fetus and its parents. Following genetic counseling, the parents has chosen to continue with the pregnancy. CONCLUSION The fetus was diagnosed with Miller-Dieker syndrome due to the de novo deletion at 17p13.3. Ventriculomegaly may be an important indicator for prenatal ultrasonography in fetuses with MDS.
Pregnancy ; Female ; Humans ; Classical Lissencephalies and Subcortical Band Heterotopias ; Comparative Genomic Hybridization ; DNA Copy Number Variations ; Fetus ; Hydrocephalus ; Prenatal Diagnosis ; Chromosome Deletion

OBJECTIVE: To explore the genetic etiology of two patients with developmental delay and intellectual disability. METHODS: Two children who were respectively admitted to Henan Provincial People's Hospital on August 29, 2021 and August 5, 2019 were selected as the study subjects. Clinical data were collected, and array comparative genomic hybridization (aCGH) was carried out on the children and their parents for the detection of chromosomal microduplication/microdeletions. RESULTS: Patient 1 was a 2-year-and-10-month female and patient 2 was a 3-year-old female. Both children had featured developmental delay, intellectual disability, and abnormal findings on cranial MRI. aCGH revealed that patient 1 has harbored arr[hg19] 6q14.2q15(84621837_90815662)×1, a 6.19 Mb deletion at 6q14.2q15, which encompassed ZNF292, the pathogenic gene for Autosomal dominant intellectual developmental disorder 64. Patient 2 has harbored arr[hg19] 22q13.31q13.33(46294326_51178264)×1, a 4.88 Mb deletion at 22q13.31q13.33 encompassing the SHANK3 gene, haploinsufficiency of which can lead to Phelan-McDermid syndrome. Both deletions were classified as pathogenic CNVs based on the guidelines of American College of Medical Genetics and Genomics (ACMG) and were not found in their parents. CONCLUSION The 6q14.2q15 deletion and 22q13-31q13.33 deletion probably underlay the developmental delay and intellectual disability in the two children, respectively. Haploinsufficiency of the ZNF292 gene may account for the key clinical features of the 6q14.2q15 deletion.
Humans ; Child ; Female ; Child, Preschool ; Intellectual Disability/genetics* ; Comparative Genomic Hybridization ; Chromosome Disorders/genetics* ; Chromosome Deletion ; Magnetic Resonance Imaging ; Chromosomes, Human, Pair 22 ; Developmental Disabilities/genetics* ; Carrier Proteins/genetics* ; Nerve Tissue Proteins/genetics*

OBJECTIVE: To analyze the 28-day survival status and influencing factors of adult patients with sepsis, providing reference for early diagnosis of sepsis prognosis and reducing sepsis mortality. METHODS: A retrospective cohort study was conducted. A total of 160 adult patients with sepsis in the department of intensive care unit of the 940th Hospital of Joint Logistic Support Force of PLA from January 2021 to December 2022 were enrolled. The general information, laboratory examination results within 24 hours after admission, clinical treatment measures, and prognosis of patients were collected. Univariate analysis and binary multivariate Logistic regression were performed to screen the risk factors that might affect the short-term prognosis of patients with sepsis. Receiver operator characteristic curve (ROC curve) was plotted to analyze the predictive value of various risk factors on the short-term death risk of sepsis patients. RESULTS: A total of 160 patients with sepsis were enrolled, of whom 91 survived in 28 days, and 69 died with a mortality of 43.12%. Compared with the survival group, the patients in the death group were older, more severe, had higher blood lactic acid (Lac) level, and had more complications such as hypertension and multiple organ dysfunction syndrome (MODS). A total of 22 related factors were statistically significant: age, acute physiology and chronic health evaluation II (APACHE II) score, sequential organ failure assessment (SOFA) score, length of hospital stay, Lac, interleukin-6 (IL-6), fibrinogen (FIB), international normalized ratio (INR), ratio of prothrombin time (PT) to healthy people, prothrombin activity (PTA), PT, thrombin time (TT), oxygenation index (PaO₂/FiO₂), aspartate aminotransferase (AST), ratio of AST to alanine amninotransferase (ALT), serum creatinine (SCr), blood urea nitrogen (BUN), site of infection, history of hypertension, concurrent MODS, implementation of continuous renal replacement therapy (CRRT), and treatment with vasoactive drugs. Combined with the results of the univariate analysis, variables that might affect the short-term prognosis of septic patients were included in the multivariate Logistic regression analysis. The results showed that APACHE II score ≥ 20 [odds ratio (OR) = 1.106, 95% confidence interval (95%CI) was 1.003-1.219], Lac ≥ 5 mmol/L (OR = 1.430, 95%CI was 1.041-1.964), combined with hypertension (OR = 13.879, 95%CI was 1.082-178.016), concurrent MODS (OR = 98.139, 95%CI was 18.252-527.672) was an independent risk factor for 28-day death in adult septic patients (all P < 0.05). ROC curve analysis showed that the combination of the four indicators including APACHE II score, Lac, combined with hypertension, concurrent MODS, had predictive value for short-term outcomes in patients with sepsis. The area under the ROC curve (AUC) was higher than that of the 4 indicators alone [AUC (95%CI): 0.952 (0.918-0.986) vs. 0.745 (0.670-0.820), 0.816 (0.748-0.883), 0.608 (0.518-0.699), 0.868 (0.810-0.927)], the sensitivity was 94.2%, and the specificity was 90.1%. CONCLUSIONS APACHE II score within 24 hours, Lac, combined with hypertension, and concurrent MODS are independent risk factors for short-term mortality in adult septic patients in ICU. The combination of these indicators can make meaningful early clinical judgments on the short-term prognosis of septic patients, thereby reducing the mortality.
Adult ; Humans ; Retrospective Studies ; ROC Curve ; Sepsis/diagnosis* ; Prognosis ; Intensive Care Units ; Risk Factors ; Hypertension