Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.

OBJECTIVE: To observe the effects of Tiaoshu Anshen (regulating the hinge and calming the mind) acupuncture on sleep quality and serum levels of 5-hydroxytryptamine (5-HT) and dopamine (DA) in patients with chronic insomnia. METHODS: A total of 58 patients with chronic insomnia were randomly divided into an acupuncture group and a medication group, 29 cases in each group. Tiaoshu Anshen acupuncture was applied at Baihui (GV20) and bilateral Shenmen (HT7), Sanyinjiao (SP6), Benshen (GB13) in the acupuncture group, once a day, 1-day interval was taken after 6 consecutive days of treatment. Estazolam tablet was given orally before bed in the medication group, 1 mg each time. The 4-week treatment was required in both groups. Before and after treatment, the sleep quality was assessed by Pittsburgh sleep quality index (PSQI) and polysomnography (PSG), the serum levels of 5-HT and DA were detected by ELISA. RESULTS: After treatment, the item scores and total scores of PSQI were decreased compared with those before treatment in the two groups (P<0.05); in the acupuncture group, the scores of sleep quality, sleep latency, sleep time, sleep efficiency, sleep disorders and total score of PSQI were lower than those in the medication group (P<0.05). After treatment, the total sleep time (TST) was prolonged (P<0.05), the sleep latency (SL) and wake after sleep onset (WASO) were shortened (P<0.05), the sleep efficiency (SE%), percentage of non-rapid eye movement stage 3 (N3%), percentage of rapid eye movement stage (REM%) and serum levels of 5-HT were increased (P<0.05) compared with those before treatment; the percentage of non-rapid eye movement stage 1 (N1%), percentage of non-rapid eye movement stage 2 (N2%) and serum levels of DA were decreased (P<0.05) compared with those before treatment in the two groups. After treatment, in the acupuncture group, TST was longer, while SL and WASO were shorter than those in the medication group (P<0.05), SE%, N3%, REM% and serum level of 5-HT were higher, while N1%, N2% and serum level of DA were lower than those in the medication group (P<0.05). CONCLUSION Tiaoshu Anshen acupuncture may improve the sleep quality by regulating the serum neurotransmitter levels i.e. 5-HT and DA in patients with chronic insomnia.
Humans ; Sleep Initiation and Maintenance Disorders/physiopathology* ; Male ; Acupuncture Therapy ; Female ; Middle Aged ; Adult ; Serotonin/blood* ; Sleep Quality ; Acupuncture Points ; Dopamine/blood* ; Aged ; Neurotransmitter Agents/blood* ; Young Adult

OBJECTIVES: To investigate the clinical sub-phenotype (SP) of pediatric acute kidney injury (AKI) and their association with clinical outcomes. METHODS: General status and initial values of laboratory markers within 24 hours after admission to the pediatric intensive care unit (PICU) were recorded for children with AKI in the derivation cohort (n=650) and the validation cohort (n=177). In the derivation cohort, a least absolute shrinkage and selection operator (LASSO) regression analysis was used to identify death-related indicators, and a two-step cluster analysis was employed to obtain the clinical SP of AKI. A logistic regression analysis was used to develop a parsimonious classifier model with simplified metrics, and the area under the curve (AUC) was used to assess the value of this model. This model was then applied to the validation cohort and the combined derivation and validation cohort. The association between SPs and clinical outcomes was analyzed with all children with AKI as subjects. RESULTS: In the derivation cohort, two clinical SPs of AKI (SP1 and SP2) were identified by the two-step cluster analysis using the 20 variables screened by LASSO regression, namely SP_d1 group (n=536) and SP_d2 group (n=114). The simplified classifier model containing eight variables (P<0.05) had an AUC of 0.965 in identifying the two clinical SPs of AKI (P<0.001). The validation cohort was clustered into SP_v1 group (n=156) and SP_v2 group (n=21), and the combined derivation and validation cohort was clustered into SP1 group (n=694) and SP2 group (n=133). After adjustment for confounding factors, compared with the SP1 group, the SP2 group had significantly higher incidence rates of multiple organ dysfunction syndrome and death during the PICU stay (P<0.001), and SP2 was significantly associated with the risk of death within 28 days after admission to the PICU (P<0.001). CONCLUSIONS This study establishes a parsimonious classifier model and identifies two clinical SPs of AKI with different clinical features and outcomes.The SP2 group has more severe disease and worse clinical prognosis.
Humans ; Acute Kidney Injury/diagnosis* ; Prognosis ; Male ; Female ; Child ; Child, Preschool ; Phenotype ; Infant ; Logistic Models ; Adolescent

Objective:To establish a three-dimensional finite element model of the nose, simulate and analyze the deformation of nasal tissue caused by different focal points, traction directions, and modes, provide the theoretical basis for the effectiveness of physical traction therapy, and guide the clinical selection of more efficient physical traction therapy methods.Methods:A finite element model of the nose was established by ANSYS Workbench 19.2 software based on image data obtained from CT scans of a 29-year-old male volunteer with normal nasal appearance in Peking University Third Hospital. Two focal points, the nasal tip, and the nasal columella, were selected, and three force directions, parallel to the forward, forward and down 30°, forward and down 60°, were applied. The deformation caused by different traction conditions on the skin, lining, and soft bone parts, as well as the four anatomical landmarks of the nasal tip, nasal root, the midpoint of the nasal columella, and the nasal base, were compared. The deformation produced by 10 minutes of continuous pulling and 10 times 1-minute pulse pulling were compared under the same pulling conditions. The deformations generated by two types of pulling modes within a 24-hour cycle: a single 1-hour cycle and 6 intermittent 10-minute cycles, were compared.Results:All traction conditions resulted in deformation of the nasal model, with the maximum deformation of the nasal tissue obtained by pulling forward and downward at 60° (4.632 9 mm) which was greater than other traction conditions (0.825 0-3.105 0 mm). The maximum deformation value was located near the nasion of the model’s skin layer. The deformation obtained by 10 minutes of continuous pulling (0.176 6 mm) was slightly greater than that obtained by 10 times of 1-minute pulse pulling (0.176 5 mm). Within 24 hours, the final deformation of multiple intermittent pulling modes (0.019 0 mm) was greater than that of a single pulling mode (0.004 3 mm).Conclusion:Physical traction can effectively deform the skin and soft tissue of the nose, and the most efficient operation is to continuously pinch the tip of the nose for a short period and apply tension parallel to the back of the nose downwards, repeating every a few hours.

Objective:To establish a three-dimensional finite element model of the nose, simulate and analyze the deformation of nasal tissue caused by different focal points, traction directions, and modes, provide the theoretical basis for the effectiveness of physical traction therapy, and guide the clinical selection of more efficient physical traction therapy methods.Methods:A finite element model of the nose was established by ANSYS Workbench 19.2 software based on image data obtained from CT scans of a 29-year-old male volunteer with normal nasal appearance in Peking University Third Hospital. Two focal points, the nasal tip, and the nasal columella, were selected, and three force directions, parallel to the forward, forward and down 30°, forward and down 60°, were applied. The deformation caused by different traction conditions on the skin, lining, and soft bone parts, as well as the four anatomical landmarks of the nasal tip, nasal root, the midpoint of the nasal columella, and the nasal base, were compared. The deformation produced by 10 minutes of continuous pulling and 10 times 1-minute pulse pulling were compared under the same pulling conditions. The deformations generated by two types of pulling modes within a 24-hour cycle: a single 1-hour cycle and 6 intermittent 10-minute cycles, were compared.Results:All traction conditions resulted in deformation of the nasal model, with the maximum deformation of the nasal tissue obtained by pulling forward and downward at 60° (4.632 9 mm) which was greater than other traction conditions (0.825 0-3.105 0 mm). The maximum deformation value was located near the nasion of the model’s skin layer. The deformation obtained by 10 minutes of continuous pulling (0.176 6 mm) was slightly greater than that obtained by 10 times of 1-minute pulse pulling (0.176 5 mm). Within 24 hours, the final deformation of multiple intermittent pulling modes (0.019 0 mm) was greater than that of a single pulling mode (0.004 3 mm).Conclusion:Physical traction can effectively deform the skin and soft tissue of the nose, and the most efficient operation is to continuously pinch the tip of the nose for a short period and apply tension parallel to the back of the nose downwards, repeating every a few hours.

Objective To clarify and analyze the reasons for failure in screening healthy menopausal female in a bioequivalence trial.Methods To summarize and clarify the data of 185 healthy menopausal female subjects participating in a bioequivalence trial of estradiol valerate conducted,and summarize the reasons for screening failure.Results The main reasons for screening failure include laboratory tests(32.04％),gynecological transvaginal color Doppler ultrasound(16.57％),vital signs(14.36％),chest computed tomography(CT,11.60％),and medical history/medication history(7.73％).Conclusion Screening failures in healthy menopausal female subjects were characterized mainly by abnormal gynecological transvaginal color Doppler ultrasound and non-compliance with basal hormone levels compared to generally healthy subjects.

Objective To investigate the correlation of CEP192 expression with prognosis of gastric cancer and biological behaviors of gastric cancer cells.Methods Public databases and clinical tissue samples were used to examine CEP192 expression level in gastric cancer.Kaplan-Meier survival curves,univariate and multivariate Cox regression analyses,ROC curves and bioinformatics analyses were used to explore the risk factors affecting the 5-year postoperative survival,the correlation of CEP192 expression level with the patients'survival,and its biological role in gastric cancer development.In gastric cancer MGC-803 cells with lentivirus-mediated CEP192 interference or overexpression,cell proliferation and expressions of PLK1,CDK1 and Cyclin B1 were examined with CCK-8 assay and Western blotting.The effects of CEP192 knockdown or overexpression on tumorigenesis of MGC-803 cells was observed in nude mice,and the expressions of PLK1,CDK1 and Cyclin B1 in the xenografts were detected.Results CEP192 was highly expressed in gastric cancer and associated with poor prognosis of the patients(P<0.05).High expression of CEP192,CEA≥5 ng/mL,CA199≥37 IU/mL,T3-4 stage,and N2-3 stage were independent risk factors affecting the patients'5-year postoperative survival(P<0.05).Bioinformatics analyses suggested that CEP192 was involved in several vital biological processes and positively regulated cell cycle progression.In MGC-803 cells,CEP192 knockdown significantly inhibited cell proliferation and lowered the expression levels of PLK1,CDK1,and Cyclin B1,while its overexpression produced the opposite effects.In the nude mouse models,CEP192 knockdown resulted in lowered tumorigenic potential of MGC-803 cells and decreased protein levels of PLK1,CDK1,and Cyclin B1 in the xenografts,while CEP192 overexpression in MGC-803 cells caused the opposite changes.Conclusion CEP192 overexpression is correlated with unfavorable outcomes of gastric cancer patients and promotes gastric cell proliferation by regulating the key proteins during G2/M phase transition.

Objective To investigate the correlation of CEP192 expression with prognosis of gastric cancer and biological behaviors of gastric cancer cells.Methods Public databases and clinical tissue samples were used to examine CEP192 expression level in gastric cancer.Kaplan-Meier survival curves,univariate and multivariate Cox regression analyses,ROC curves and bioinformatics analyses were used to explore the risk factors affecting the 5-year postoperative survival,the correlation of CEP192 expression level with the patients'survival,and its biological role in gastric cancer development.In gastric cancer MGC-803 cells with lentivirus-mediated CEP192 interference or overexpression,cell proliferation and expressions of PLK1,CDK1 and Cyclin B1 were examined with CCK-8 assay and Western blotting.The effects of CEP192 knockdown or overexpression on tumorigenesis of MGC-803 cells was observed in nude mice,and the expressions of PLK1,CDK1 and Cyclin B1 in the xenografts were detected.Results CEP192 was highly expressed in gastric cancer and associated with poor prognosis of the patients(P<0.05).High expression of CEP192,CEA≥5 ng/mL,CA199≥37 IU/mL,T3-4 stage,and N2-3 stage were independent risk factors affecting the patients'5-year postoperative survival(P<0.05).Bioinformatics analyses suggested that CEP192 was involved in several vital biological processes and positively regulated cell cycle progression.In MGC-803 cells,CEP192 knockdown significantly inhibited cell proliferation and lowered the expression levels of PLK1,CDK1,and Cyclin B1,while its overexpression produced the opposite effects.In the nude mouse models,CEP192 knockdown resulted in lowered tumorigenic potential of MGC-803 cells and decreased protein levels of PLK1,CDK1,and Cyclin B1 in the xenografts,while CEP192 overexpression in MGC-803 cells caused the opposite changes.Conclusion CEP192 overexpression is correlated with unfavorable outcomes of gastric cancer patients and promotes gastric cell proliferation by regulating the key proteins during G2/M phase transition.

Cancer is one of the deadliest diseases affecting the health of human beings. With limited therapeutic options available, complementary and alternative medicine has been widely adopted in cancer management and is increasingly becoming accepted by both patients and healthcare workers alike. Chinese medicine characterized by its unique diagnostic and treatment system is the most widely applied complementary and alternative medicine. It emphasizes symptoms and ZHENG (syndrome)-based treatment combined with contemporary disease diagnosis and further stratifies patients into individualized medicine subgroups. As a representative cancer with the highest degree of malignancy, pancreatic cancer is traditionally classified into the "amassment and accumulation". Emerging perspectives define the core pathogenesis of pancreatic cancer as "dampness-heat" and the respective treatment "clearing heat and resolving dampness" has been demonstrated to prolong survival in pancreatic cancer patients, as has been observed in many other cancers. This clinical advantage encourages an exploration of the essence of dampness-heat ZHENG (DHZ) in cancer and investigation into underlying mechanisms of action of herbal formulations against dampness-heat. However, at present, there is a lack of understanding of the molecular characteristics of DHZ in cancer and no standardized and widely accepted animal model to study this core syndrome in vivo. The shortage of animal models limits the ability to uncover the antitumor mechanisms of herbal medicines and to assess the safety profile of the natural products derived from them. This review summarizes the current research on DHZ in cancer in terms of the clinical aspects, molecular landscape, and animal models. This study aims to provide comprehensive insight that can be used for the establishment of a future standardized ZHENG-based cancer animal model.
Animals ; Humans ; Medicine, Chinese Traditional ; Hot Temperature ; Pancreatic Neoplasms/therapy* ; Models, Animal ; Syndrome

Based on the long bud stage phenotype of a new Lonicera japonica Flos variety "Huajin 6", using "Huajin 6" and "Da Mao Hua" as materials, probing the mechanism of its phenotype formation. Detection of endogenous Jasmonic acid hormones (JAs) content; the genes related to jasmonic acid (JA) synthesis were identified by transcriptome analysis of Lonicera japonica; flower buds and flowers of "Huajin 6" and "Da Mao Hua" were collected at different periods, and the qRT-PCR (quantitative real-time PCR) technique was used to analyze the trend of the expression of synthesis-related enzyme genes in Lonicera japonica Flos during the bud stage. The study found that the content of JAs in "Huajin 6" Lonicera japonica Flos was significantly lower than that in "Da Mao Hua"; applying exogenous methyl-jasmonate (MeJA) to "Huajin 6" can restore its flowering phenotype, making it close to wild type Lonicera japonica Flos; there are significant differences in the expression of two allene oxide synthase genes (AOS), three lipoxygenase genes (LOX), and two allene oxide cyclase genes (AOC) in the flowers and buds of "Huajin 6" and "Da Mao Hua" at different periods. It is hypothesized that the low expression of JA synthesis-related enzyme genes in " Huajin 6" leads to the blockage of JA synthesis, which causes the formation of the long bud phenotype. This study laid a certain foundation for the genetic breeding of Lonicera japonica, provided a new idea for the improvement of Lonicera japonica varieties, and provided a reference for the study of JAs in plant flower organs.