AIM: To evaluate the expression of tumor necrosis factor-α(TNF-α)in the iris and ciliary body of Wistar rats in the endotoxin-induced uveitis(EIU), and the effect of Xuebijing injection on its expression.METHODS:A total of 65 Wistar rats were randomly divided into three groups: Group A(normal saline, n=5), Group B(normal saline+endotoxin-injected, n=30), and Group C(Xuebijing+endotoxin-injected, n=30). The EIU model was induced in Wistar rats of the groups B and C by injecting LPS into the plantar surfaces of the hind feet, and normal saline(15 mL/kg)or Xuebijing(15 mL/kg)were intraperitoneally administered 30 min before LPS administration. The rats of the groups B and C were further divided into 6 subgroups after LPS injection, including 6, 12, 18, 24, 48, and 72 h subgroups, with 5 rats in each group. Furthermore, the intraocular inflammation of the rats was observed at each time above, the number of infiltrating cells in the aqueous humor was counted, and the pathological changes were observed in the iris and ciliary body of rats using hematoxylin and eosin(HE)staining. TNF-α expression in iris and ciliary tissue at different postoperative time points was evaluated using immunohistochemistry.RESULTS: Clinical observations indicated no signs of uveitis in the group A, signs of uveitis were observed in the group B. Both iris symptoms and damage were significantly reduced in the group C compared to the group B(P<0.01). Cell counts in the aqueous humor revealed no inflammatory cells in the group A, while the number of aqueous humor cells in the group C was significantly reduced compared to Group B(P<0.01). HE staining revealed no cellular infiltration in the group A. In the group B, some cellular infiltration was observed in the eyes at 6 h post-LPS exposure. The number of infiltrating cells increased over time, peaked at 24 h, and gradually declined thereafter. In the group C, cell infiltration was not obvious at 6 h, few at 24 h, and nearly disappeared by 48 h. Immunohistochemical staining showed higher TNF-α expression in the ciliary body and iris in the group B than in the group A(P<0.01). Compared to the group C, TNF-α expression in the group B was significantly upregulated following LPS injection(P<0.01).CONCLUSION:TNF-α expression was elevated in EIU rats, and there was a positive correlation between its mean optical density ratio and inflammation degree. Moreover, Xuebijing injection could alleviate inflammation response through the reduction of TNF-α levels.

Objective To examine the compliance of and factors affecting target attainment of serum trough concentration of norvancomycin in ICU patients,and the effects of different trough concentrations on clinical efficacy and renal impairment.Methods Adult patients admitted to the Department of Critical Care Medicine of Huangshan People's Hospital and receiving intravenous infusion of norvancomycin from January 2020 to December 2022 were included.The dosing regimens and steady-state trough concentrations of norvancomycin were analyzed.The clinical efficacy and renal impairment were compared between different trough concentration levels.The compliance of trough concentration in critically ill patients with different renal functions was examined.Logistic regression analysis was performed to profile the factors possibly affecting the trough concentration of norvancomycin.Results A total of 97 patients were included.The target serum trough concentration(10-20 mg/L)was reached in only 33.0％(32/97)of the critically ill patients.The serum trough concentration was below the target in 51.5％(50/97)of the patients,above the target in 15.5％of the patients.The clinical cure rate and incidence of renal impairment were significantly different among the three groups of patients with different trough concentrations(P＜0.05).The compliance with target serum trough concentration varied with different renal function tests(P＜0.05).Augmented renal clearance and normal renal function were associated with trough concentrations lower than the target.As renal dysfunction got worse,serum trough concentration was more probably higher than the target(P＜0.05).Univariate analysis showed that daily dose,age,gender,height,weight,body mass index(BMI),APACHE Ⅱ score,sequential organ failure assessment(SOFA)score,blood creatinine,urea nitrogen,procalcitonin,concomitant septic shock,and use of norepinephrine were significantly correlated with trough concentrations of norvancomycin(P＜0.05).Multivariate logistic regression analysis indicated that age,SOFA score,blood urea nitrogen,gender,and norepinephrine use were independent factors affecting the serum trough concentration of norvancomycin(P＜0.05).Conclusions The serum trough concentration of norvancomycin varied with renal function states in ICU patients.It is difficult to achieve the steady-state target trough concentration(10-20 mg/L).The clinical cure rate is lower when the trough concentration is lower than the target.As the trough concentration increases,the incidence of renal impairment increases.Age,SOFA score,urea nitrogen,gender,and norepinephrine use are independent factors affecting the serum trough concentrations of norvancomycin.

ω-transaminases are able to catalyze the reversible transfer of amino groups between diverse amino compounds (such as amino acids, alkyl amines, aromatic amines) and carbonyl compounds (such as aldehydes, ketones, ketoacids). ω-transaminases exhibit great application prospects in the field of chiral amine biosynthesis because of their desirable properties, such as wide range of substrates, high stereoselectivity, and mild catalytic conditions. It is therefore important for China to develop efficient, specific, and environment-friendly chiral amine production technologies with independent intellectual property rights, which is of great significance for the development of pharmaceutical, pesticide, and material industries. This review systematically summarizes the Chinese patents regarding ω-transaminase filed by Chinese institutions in the recent decade. The development of ω-transaminase resource, enzymatic property improvement by protein engineering, application in chiral amine synthesis, and development of production technologies are elaborated. This review will shed light on further basic and application studies of ω-transaminase.
Transaminases/genetics* ; Amino Acids ; China ; Aldehydes ; Amines

Sialyllactose is one of the most abundant sialylated oligosaccharides in human milk oligosaccharides (HMOs), which plays an important role in the healthy development of infants and young children. However, its efficient and cheap production technology is still lacking presently. This study developed a two-step process employing multiple-strains for the production of sialyllactose. In the first step, two engineered strains, E. coli JM109(DE3)/ pET28a-BT0453 and JM109(DE3)/pET28a-nanA, were constructed to synthesize the intermediate N-acetylneuraminic acid. When the ratio of the biomass of the two engineered strains was 1:1 and the reaction time was 32 hours, the maximum yield of N-acetylneuraminic acid was 20.4 g/L. In the second step, E. coli JM109(DE3)/ pET28a-neuA, JM109(DE3)/ pET28a-nst and Baker's yeast were added to the above fermentation broth to synthesize 3'-sialyllactose (3'-SL). Using optimal conditions including 200 mmol/L N-acetyl-glucosamine and lactose, 150 g/L Baker's yeast, 20 mmol/L Mg²⁺, the maximum yield of 3'-SL in the fermentation broth reached 55.04 g/L after 24 hours of fermentation and the conversion rate of the substrate N-acetyl-glucosamine was 43.47%. This research provides an alternative technical route for economical production of 3'-SL.
Child ; Humans ; Child, Preschool ; N-Acetylneuraminic Acid ; Escherichia coli/genetics* ; Lactose ; Fermentation ; Saccharomyces cerevisiae ; Oligosaccharides ; Glucosamine

Objective:To investigate the expression of serum human phosphatidylethanolamine-binding protein 4 (hPEBP4) in patients with multiple myeloma (MM) and its clinical significance.Methods:A total of 59 symptomatic MM patients admitted to West Branch of Beijing Chaoyang Hospital from September 2016 to September 2018 were selected as the research objects. According to the CRAB symptoms [elevated serum calcium (C), kidney injury (R), anemia (A), bone lesions (B)], all patients were divided into 2 groups, including the active group of 44 patients with CRAB symptoms, and the response group of 15 patients who achieved at least partial remission after chemotherapy and symptom relief of CRAB. According to the degree of bone lesions (BL), 30 patients with severe bone-related events were grouped as the severe bone lesions (SBL) group, and 14 patients were grouped as the non-severe bone lesions (NSBL) group. According to the revised international prognostic staging system (R-ISS), patients in the active group were divided into three subgroups: stage Ⅰ, stage Ⅱ, and stage Ⅲ, including 26, 11 and 7 patients, respectively. A total of 15 healthy examination people whose gender and age matched those of the patients were treated as the healthy control group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression levels of hPEBP4, tumor necrosis factor ligand superfamily member 14 (LIGHT/TNFSF14) and activin A of patients in different groups. Pearson was used to analyze the relationship of the expressions of multiple factors in the active group. The optimal cut-off value of multiple factors diagnosing MM was determined by using receiver operating characteristic (ROC) curve, and according to the cut-off value, the differences in overall survival (OS) of patients with different stratification were compared.Results:In the active group, the respond group, the healthy control group, the level of hPEBP4 was (1.48±0.64) μg/L, (1.49±0.75) μg/L, (0.31±0.10) μg/L, respectively; the level of LIGHT/TNFSF14 was (169±112) ng/L, (256±132) ng/L, (44±27) ng/L,respectively; the level of activin A was (383±266) ng/L, (223±79) ng/L, (234±85) ng/L, respectively; and the differences were statistically significant (all P<0.05). In the active group, the level of hPEBP4 was (1.06±0.60) μg/L, (1.15±0.50) μg/L, (1.73±0.68) μg/L, respectively in patients with stage R-ISSⅠ, R-ISSⅡ and R-ISS Ⅲ, and the difference was statistically significant ( F=3.287, P=0.032). The level of activin A was (219±55) ng/L, (247±117) ng/L, (450±215) ng/L, respectively among patients in stage R-ISSⅠ, R-ISSⅡ, R-ISS Ⅲ, and the level of activin A in stage R-ISS Ⅲ was higher than that in stage R-ISSⅠand R-ISSⅡ (all P < 0.05). The levels of LIGHT/TNFSF14 and activin A of SBL patients were higher than those of NSBL patients [(174±101) ng/L vs. (98±53) ng/L; (467±238) ng/L vs. (189±71) ng/L, all P < 0.05]. The level of hPEBP4 was positively correlated with the levels of M protein ( r=0.694, P < 0.01) and activin A ( r=0.252, P < 0.01) of IgG patients in the active group. ROC curve analysis showed that the optimal cut-off value of hPEBP4, LIGHT/TNFSF14, activin A diagnosing MM was 1.04 μg/L, 97.0 μg/L, 156.2 ng/L. The median overall survival (OS) time of patients with hPEBP4 >1.04 μg/L and hPEBP4 ≤ 1.04 μg/L was 57 months (95% CI 22-92 months) and not reached, respectively, and the difference was statistically significant ( P < 0.05); while the median OS time of patients with activin A ≥ 156.2 ng/L and activin A < 156.2 ng/L was 61 months (95% CI 24-98 months) and not reached, respectively, and the difference was statistically significant ( P < 0.05). Conclusions:High expression level of hPEBP4 is related with the progression of MM. It is positively related with the level of M protein and negatively with the OS of MM patients. It is suggested that hPEBP4 may be used as an important marker to judge disease progression and tumor burden in MM. LIGHT/TNFSF14 and activin A cooperate with hPEBP4 to participate in the pathological processes of tumor microenvironment of MM.

Chromosome microarray analysis (CMA) has become the first-tier testing for chromosomal abnormalities and copy number variations (CNV). This review described the clinical validation of CMA, the development and updating of technical standards and guidelines and their diagnostic impacts. The main focuses were on the development and updating of expert consensus, practice resources, and a series of technical standards and guidelines through systematic review of case series with CMA application in the literature. Expert consensus and practice resource supported the use of CMA as the first-tier testing for detecting chromosomal abnormalities and CNV in developmental and intellectual disabilities, multiple congenital anomalies and autism. The standards and guidelines have been applied to pre- and postnatal testing for constitutional CNV and tumor testing for acquired CNV. CMA has significantly improved the diagnostic yields but still needs to overcome its technical limitations and face challenges of new technologies. Guiding and governing CMA through expert consensus, practice resource, standards and guidelines in the United States has provided effective and safe diagnostic services to patients and their families, reliable diagnosis on related genetic diseases for clinical database and basic research, and references for clinical translation of new technologies.
Child ; Chromosome Aberrations ; Chromosomes ; DNA Copy Number Variations ; Developmental Disabilities/genetics* ; Humans ; Intellectual Disability/genetics* ; Microarray Analysis ; United States

A 49-year-old woman was admitted to hospital with intermittent dizziness and fatigue for 7 years. The symptoms were aggravated and accompanied by bone pain for more than 4 months. She was referred to our hospital. Laboratory tests and imaging findings suggested that acquired Fanconi Syndrome (FS) was associated with smoldering multiple myeloma (MM). Renal biopsy and electron microscopy confirmed the diagnosis of proximal light chain tubular disease (LCPT). LCPT causes proximal tubular dysfunction, which is characterized by the cytoplasmic crystal deposition usually kappa monoclonal light chain in the proximal tubule. MM with FS and LCPT is less common in clinical practice because it is difficult to diagnose. This is a typical case focusing on the differential diagnosis of monoclonal gammopathy of renal significance(MGRS) such as LCPT and plasma cells diseases.

Objective:To explore the accuracy of determination of the pathogen in children with lower respiratory infection according to sputum properties and the selection of corresponding antibiotics.To analyze the relationship between children's age, length of hospital stays and sputum culture positive rate.Methods:From March 2015 to November 2016, 300 children with lower respiratory infection in the Seventh Affiliated Hospital of Guangxi Medical University were selected.The isolated sputums through the nasal cavity were cultured, the sputum properties and antibiotics used at the beginning and 3-4 days later(after sputum culture) were recorded.Results:The sputum culture showed that 110 of 300 cases with lower respiratory infection were positive.Main pathogens were streptococcus pneumoniae, E.coli, staphylococcus aureus, klebsiella pneumoniae subspecies.Compared with the children with positive sputum culture, for white phlegm sputum and yellow phlegm sputum, the children with sputum culture negative had higher accuracy of antibiotic use(white phlegm sputum: 64.4%, yellow phlegm sputum: 57.1%), the difference was statistically significant (χ 2=36.04, P<0.01). The bacteria positive rate and multiple resistance rate had no statistically significant differences in length of treatment time prior to admission(all P>0.05). However, the bacteria positive rate (hospital stay ≤7 d: 24.7%, hospital stay>7 d: 48.1%, χ 2=17.66) and multiple resistance rate (hospital stay≤7 d: 13.9%, hospital stay>7 d: 35.1%; χ 2=5.40) had statistically significant differences in the length of hospitalization(all P<0.05). The bacteria positive rates in different age groups were as follows: 1-6 months: 48.1%, >6-36 months: 28.3%, >36 months: 25.0%.Compared with the two others, the positive rate of 1-6 months was higher and had statistically significant difference (χ 2=13.64, P<0.05). Conclusion:Accuracy of antibiotics use has a certain relationship with sputum color or properties and the result of sputum culture, for white phlegm sputum and yellow phlegm sputum, the accuracy of antibiotics use is higher.Once identified pathogen, sensitive antibiotics should be adopted.Probably, bacteria positive rate and multiple resistance rate have a certain relationship with the length of hospital stay and age.

Objective:To explore the relationship between dietary quality and the risk of diabetes and prediabetes among urban women based on alternative healthy eating index-2010（AHEI-2010).Methods:From March to July 2016, a total of 1 061 female residents from 8 cities, Beijing, Guangzhou, Chengdu, Lanzhou, Xuchang, Jilin, Wuhu and Chenzhou in China, were selected by using the multi-stage sampling method. The basic characteristics of all participants were collected by using self-made questionnaire, and the dietary status of the subjects was collected by using 24-hour dietary recall method. AHEI-2010 scores were calculated and the diet was divided into low, medium and high dietary quality group according to the third quartile. All participants received physical examination, systolic blood pressure test, fasting blood glucose test and glycosylated hemoglobin test. Unconditional logistic regression model was used to analyze the relationship between dietary quality and the risk of diabetes and prediabetes.Results:The age of all participants was（48.0±17.5) years old. The median（ P25, P75) of AHEI-2010 score was 46.52（40.04, 54.88). The prevalence of diabetes and prediabetes were 13.3%（141 cases) and 50.0%（530 cases), respectively. Unconditional logistic regression analysis showed that after adjusting for age, city, education, diet control, physical activity, energy intake and waist circumference, compared with the low diet quality group, the OR（95% CI) values of medium, high dietary quality group and diabetes risk were 0.96（0.57-1.62) and 0.63（0.36-1.09), respectively; the OR（95% CI) values of medium, high dietary quality group and prediabetes risk were 0.93（0.64-1.35) and 1.28（0.87-1.88), respectively. Conclusion:The dietary quality of urban women based on AHEI-2010 evaluation is not related to the risk of diabetes and prediabetes.

Objective:To explore the relationship between dietary quality and the risk of diabetes and prediabetes among urban women based on alternative healthy eating index-2010（AHEI-2010).Methods:From March to July 2016, a total of 1 061 female residents from 8 cities, Beijing, Guangzhou, Chengdu, Lanzhou, Xuchang, Jilin, Wuhu and Chenzhou in China, were selected by using the multi-stage sampling method. The basic characteristics of all participants were collected by using self-made questionnaire, and the dietary status of the subjects was collected by using 24-hour dietary recall method. AHEI-2010 scores were calculated and the diet was divided into low, medium and high dietary quality group according to the third quartile. All participants received physical examination, systolic blood pressure test, fasting blood glucose test and glycosylated hemoglobin test. Unconditional logistic regression model was used to analyze the relationship between dietary quality and the risk of diabetes and prediabetes.Results:The age of all participants was（48.0±17.5) years old. The median（ P25, P75) of AHEI-2010 score was 46.52（40.04, 54.88). The prevalence of diabetes and prediabetes were 13.3%（141 cases) and 50.0%（530 cases), respectively. Unconditional logistic regression analysis showed that after adjusting for age, city, education, diet control, physical activity, energy intake and waist circumference, compared with the low diet quality group, the OR（95% CI) values of medium, high dietary quality group and diabetes risk were 0.96（0.57-1.62) and 0.63（0.36-1.09), respectively; the OR（95% CI) values of medium, high dietary quality group and prediabetes risk were 0.93（0.64-1.35) and 1.28（0.87-1.88), respectively. Conclusion:The dietary quality of urban women based on AHEI-2010 evaluation is not related to the risk of diabetes and prediabetes.