Non-specific nucleases(NSNs) are widely distributed in animals, plants, bacteria, fungi, and viruses, and can nonspecifically degrade almost all forms of nucleic acids, including single-stranded, double-stranded, linear, circular, and supercoiled DNA and RNA with no requirement on nucleic acid sequences. NSNs are broadly used in various steps of the biopharmaceutical industry and biotechnology that require the removal of nucleic acids. Currently, only a few non-restrictive nucleases have been studied in detail from both structural and functional aspects. In recent years, in addition to traditional fields such as vaccines, the continuous development of emerging fields such as cell and gene therapy has broadened the scope of application of nucleases on the one hand, but also posed new challenges in terms of reaction temperature, salt concentration, and metal chelator tolerance. This paper summarizes different species of NSNs and analyzes their homology and structural catalytic characteristics. Based on a comprehensive comparison of the basic characteristics of representative NSNs such as Benzonase~?, Denarase~?, and HL-SAN~(TM), this paper summarizes various factors that need to be considered for recombinant nucleases used in the biopharmaceutical industry, and prospects for the future research and development trends of NSNs.

Objective·To observe the effects of gingipain extract on the biological characteristics of oral squamous cell carcinoma cell HN6.Methods·The HN6 cell line was selected,cultivated,and divided into different groups based on the protein concentration of gingipain extract from Porphyromonas gingivalis:control group,3.125 μg/mL group,6.25 μg/mL group,12.5 μg/mL group,25 μg/mL group,50 μg/mL group,and 100 μg/mL group.After 24 and 48 h of cultivation,CCK-8 assay was used to detect the effects of gingipain extract on HN6 cell proliferation activity.Subsequent experiments were divided into control group,25 μg/mL group and 50 μg/mL group.Flow cytometry was used to examine the effects of gingipain extract on cell cycle.Scratch assay and Transwell assay were performed to evaluate cell migration and invasion ability.Real-time PCR(RT-PCR)and Western blotting were used to measure the expression of E-cadherin and N-cadherin proteins and genes in cells.Results·Stimulated with gingipain extract for 24 h,the HN6 cells showed significantly increased proliferation activity in the 25 μg/mL(P=0.025),50 μg/mL(P=0.000),and 100 μg/mL(P=0.049)groups compared to the control group.After 48 h,proliferation activity was significantly higher in the 6.25 μg/mL(P=0.024),12.5 μg/mL(P=0.006),25 μg/mL(P=0.000),50 μg/mL(P=0.000),and 100 μg/mL(P=0.000)groups compared to the control group.Cell cycle analysis revealed that,after 24 h of gingipain stimulation,the proportion of HN6 cells in the G1 phase decreased,while the proportion in the S+G2 phase significantly increased compared to the control group(25 μg/mL group:P=0.024;50 μg/mL group:P=0.001).Compared to the control group,the scratch assay demonstrated a significant increase in the percentage of scratch closure as the concentration of gingipain extract increased(P=0.001).Compared to the control group,the Transwell invasion assay showed a significant increase in the number of cells passing through the bottom of the chamber as the concentration of gingipain extract increased.RT-PCR and Western blotting results indicated that as the concentration of gingipain extract increased,the expression levels of N-cadherin mRNA and protein in HN6 cells significantly increased,while the expression levels of E-cadherin mRNA and protein significantly decreased compared to the control group.Conclusion·Gingipain extract could promote proliferation,migration,and invasion of oral squamous cell carcinoma HN6 cells.

Molecular knowledge of human gastric corpus epithelium remains incomplete. Here, by integrated analyses using single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and single-cell assay for transposase accessible chromatin sequencing (scATAC-seq) techniques, we uncovered the spatially resolved expression landscape and gene-regulatory network of human gastric corpus epithelium. Specifically, we identified a stem/progenitor cell population in the isthmus of human gastric corpus, where EGF and WNT signaling pathways were activated. Meanwhile, LGR4, but not LGR5, was responsible for the activation of WNT signaling pathway. Importantly, FABP5 and NME1 were identified and validated as crucial for both normal gastric stem/progenitor cells and gastric cancer cells. Finally, we explored the epigenetic regulation of critical genes for gastric corpus epithelium at chromatin state level, and identified several important cell-type-specific transcription factors. In summary, our work provides novel insights to systematically understand the cellular diversity and homeostasis of human gastric corpus epithelium in vivo.
Humans ; Epigenesis, Genetic ; Gastric Mucosa/metabolism* ; Chromatin/metabolism* ; Stem Cells ; Epithelium/metabolism* ; Fatty Acid-Binding Proteins/metabolism*

Pregnancy ; Female ; Humans ; Pre-Eclampsia/epidemiology* ; Prenatal Exposure Delayed Effects ; Case-Control Studies ; China/epidemiology* ; Fluorocarbons/toxicity*

Objective:To explore the association between different feeding patterns, emotional states, and salivary oxytocin (OT) levels during breastfeeding.Methods:From January to December 2019, 153 pairs of 3-month-old infants and their mothers were recruited from 4 maternal and child health hospitals in Chongqing, Liuzhou, Dalian and Hangzhou in China.Saliva samples were collected from the mothers at the first 5 minutes of feeding, 5 minutes during feeding, and 10 minutes after feeding.Edinburgh postnatal depression scale (EPDS) was used to evaluate maternal depression.Infants were divided into exclusive breastfeeding group and artificial feeding group according to feeding patterns.ELISA of salivary oxytocin was performed by ELISA kits, and the OT levels measured at the 3 time points were converted using linear interpolation.Area under the curve with respect to ground(OTAUCG) was used to represent the total concentration of salivary OT during the mother's breastfeeding.SPSS 25.0 software was used for statistical analysis.Multiple linear regression analysis and two factors analysis of variance were used to explore the association between different feeding methods, emotional state and salivary oxytocin during breastfeeding.Results:The results of the two factors analysis of variance showed that the interaction between feeding pattern and mother's emotion was not significant ( F=2.440, P=0.120), the main effect of mother's emotion was not significant ( F=0.380, P=0.539), and the main effect of feeding style was significant ( F=3.350, P=0.021). The level of OTAUCG under pure breastfeeding ((151 561.47±75 738.11) pg/mL) was higher than that under artificial feeding ((122 269.03±65 029.88) pg/mL), and the difference was statistically significant ( P=0.02). There was no statistically significant difference in OTAUCG levels between mothers with normal emotions ((146 106.37±75 106.76) pg/mL) and mothers with depressed emotions ((129 079.56±67 565.87) pg/mL) ( P=0.221). Multiple stepwise regression analysis showed that artificial feeding had a negative predictive effect on maternal salivary OT levels compared to exclusive breastfeeding( β=-0.211, t=-2.513, P=0.013). Conclusion:Feeding pattern is a factor that affects the mother's salivary OT level, and breastfeeding can improve the mother's OT level.

Objective:To prepare the hydrogel scaffolds with different concentrations of laponite and compare their osteogenic properties.Methods:The scaffolds of gelatin/sodium alginate hydrogel into which laponite was added according to the mass ratios of 0%, 1%, 2%, and 3% were assigned into groups T0, T1, T2, and T3. In each group, the compressive modulus was measured and the leaching solution for 24 h extracted to measure the ion release. Bone marrow mesenchymal stem cells (BMSCs) were cultured in the extract medium from each group and common medium (blank group) ( n=3) in the in vitro experiments to determine the expression of osteogenic genes Runt-related transcription factor 2 (Runx2), alkaline phosphatase (ALP) and type I collagen after 7 days of culture. In the in vivo experiments, the scaffolds were implanted into the femoral condyle defects in rats, and a blank group with no scaffolds was set. The bone repair in each group was evaluated by hematoxylin-eosin(HE) staining and immunohistochemical staining. Results:The compressive modulus in group T2 [(139.05±6.43) kPa] was significantly higher than that in groups T0, T1 and T3 [(68.83±3.76) kPa, (101.18±3.68) kPa and (125.40±3.28) kPa] ( P<0.05). The ion contents of lithium, magnesium and silicon released from the 24 h leaching solution in group T2 were (0.031±0.005) μg/mL, (3.047±0.551) μg/mL and (5.243±0.785) μg/mL, insignificantly different from those in group T3 ( P> 0.05) but significantly larger than those in group T1 ( P>0.05). The in vitro experiments showed that the expression levels of Runx2, ALP and type I collagen in group T2 were 1.59±0.11, 2.02±0.08 and 1.06±0.17, significantly higher than those in the other groups ( P<0.05). HE staining showed that the implanted hydrogel was tightly bound to the bone tissue. Immunohistochemical staining showed that the numbers of Runx2 and osteocalcin positive cells in group T2 were significantly higher than those in the other groups. Conclusions:With ideal biocompatibility, hydrogel scaffolds with different concentrations of laponite can slowly release the decomposed ions of lithium, magnesium and silicon to promote the osteogenic differentiation of BMSCs and the repair of bone defects in vivo. A 2% concentration of laponite in the hydrogel scaffolds may result in the best results.

Oxytocin is a neuropeptide produced primarily in the hypothalamus and plays an important role in the regulation of mammalian birth and lactation. It has been shown that oxytocin has important cardiovascular protective effects. Here we investigated the effects of oxytocin on vascular reactivity and underlying the mechanisms in human umbilical vein endothelial cells (HUVECs) in vitro and in rat aorta ex vivo. Oxytocin increased phospho-eNOS (Ser 1177) and phospho-Akt (Ser 473) expression in HUVECs in vitro and the aorta of rat ex vivo. Wortmannin, a specific inhibitor of phosphatidylinositol 3-kinase (PI3K), inhibited oxytocin-induced Akt and eNOS phosphorylation. In the rat aortic rings, oxytocin induced a biphasic vascular reactivity: oxytocin at low dose (10-9–10-8 M) initiated a vasorelaxation followed by a vasoconstriction at high dose (10-7 M). L-NAME (a nitric oxide synthase inhibitor), endothelium removal or wortmannin abolished oxytocin-induced vasorelaxation, and slightly enhanced oxytocin-induced vasoconstriction. Atosiban, an oxytocin/ vasopressin 1a receptor inhibitor, totally blocked oxytocin-induced relaxation and vasoconstriction. PD98059 (ERK1/2 inhibitor) partially inhibited oxytocin-induced vasoconstriction. Oxytocin also increased aortic phospho-ERK1/2 expression, which was reduced by either atosiban or PD98059, suggesting that oxytocin-induced vasoconstriction was partially mediated by oxytocin/V1aR activation of ERK1/2. The present study demonstrates that oxytocin can activate different signaling pathways to cause vasorelaxation or vasoconstriction. Oxytocin stimulation of PI3K/eNOS-derived nitric oxide may participate in maintenance of cardiovascular homeostasis, and different vascular reactivities to low or high dose of oxytocin suggest that oxytocin may have different regulatory effects on vascular tone under physiological or pathophysiological conditions.

ObjectiveTo automatically measure the wrist range of motion using the somatosensory capture device Azure Kinect. MethodsAzure Kinect was used to recognize the spatial coordinates of human elbow, wrist, palm, fingertip and other joint points, and the best measurement posture was determined by orthogonal experiment. Holt's double-parameter exponential smoothing method and bone length constraint method were used to smooth the joint point data to eliminate the jitter. The average angle of multiple frames was calculated through the spatial vector relationship to realize the automatic measurement of wrist joint range of motion. From May to October, 2021, 5 times × 10 groups of measurement were performed on each subject of wrist joint of five healthy subjects using the above method. ResultsThe R1 of each subject of wrist joint was between 83 and 127, and the value was in the middle. This method contrasted well with measurements made on augmented reality ruler, 2D images (r > 0.990), and the maximum error was within 1.61°. ConclusionThe wrist joint angle measurement modality using Azure Kinect is consistent to conventional measurement modality, and can meet the requirements of real-time measurement.

Objective:To explore the difference of high-risk factors between early-onset and late-onset pre-eclampsia, and to further understand high-risk factors of pre-eclampsia.Methods:Clinical data of pre-eclampsia pregnant women in 160 medical institutions in China in 2018 were retrospectively analyzed, including 8 031 cases of early-onset pre-eclampsia and 12 969 cases of late-onset pre-eclampsia. The proportion of high-risk factors, different body mass index (BMI) and age stratification between early-onset group and late-onset group were compared.Results:(1) Univariate analysis of high-risk factors: the proportions of high-risk factors in early-onset group and late-onset group were compared, and the differences were statistically significant (all P<0.05). Among them, the proportions of primipara and multiple pregnancy in early-onset group were lower than those in late-onset group, while the proportions of pregnant women with advanced age, irregular antenatal examination, obesity, family history of hypertension, pre-eclampsia, diabetes, kidney diseases, immune system diseases and assisted reproductive technology were higher than those in late-onset group. (2) Hierarchical analysis of BMI: the proportion of pregnant women with BMI≥24 kg/m 2 in early-onset group [48.2% (2 828/5 872) vs 45.5% (4 177/9 181), respectively; P=0.001] and the proportion of pregnant women with BMI ≥28 kg/m 2 in early-onset group [19.5% (1 143/5 872) vs 18.0% (1 656/9 181), respectively; P=0.028] were significantly higher than those in late-onset group. (3) Age stratification analysis: the proportion of pregnant women aged 35-39 years in the early-onset group [21.8% (1 748/8 023) vs 17.5% (2 110/12 068), respectively; P<0.01], the proportion of pregnant women 40-44 years old [6.8% (544/8 023) vs 5.4% (648/12 068), respectively; P<0.01], and the proportion of pregnant women ≥45 years old [0.7% (58/8 023) vs 0.5% (57/12 068), respectively; P=0.021] were significantly higher than those in the late-onset group. (4) Multivariate analysis: advanced age (≥35 years old), multiple pregnancy, irregular antenatal examination or transfer from other hospitals, family history of hypertension (including paternal, maternal and parental lines), previous history of pre-eclampsia, kidney diseases, immune system diseases (systemic lupus erythematosus, antiphospholipid antibody syndrome) and assisted reproductive technology pregnancy were the risk factors affecting the severity of pre-eclampsia (all P<0.05). Conclusion:Pregnant women with high risk factors such as age ≥35 years old, BMI ≥24 kg/m 2 before pregnancy, family history of hypertension, history of pre-eclampsia, chronic kidney diseases, immune diseases (mainly including systemic lupus erythematosus and antiphospholipid syndrome) and assisted reproductive technology are more likely to have early-onset pre-eclampsia.

Objective:To investigate the prevention and treatment status of venous thromboembolism (VTE) of pregnant women in general hospitals and specialized hospitals in China.Methods:This study was a cross-sectional survey about VTE prevention and treatment in 112 hospitals across China from January 1st to December 31st, 2019, including general information, resource accessibility, hospital system and strategy. According to the characteristics, the hospitals were divided into general hospital group (70 hospitals) and specialized hospital group (42 hospitals). The basic information, VTE diagnosis and treatment resources, prevention systems and the preference of the VTE risk assessment form of the two groups were compared. Logistic regression was used to analyze the proportion of VTE incidence and related factors in the two groups of hospitals.Results:(1) The median annual delivery volume of the general hospital group was significantly lower than that of the specialized hospital group (3 428 vs 9 969 cases, P<0.01), the diagnostic accessibility of CT pulmonary angiography (CTPA) was significantly higher than that of the specialized hospital [92.9% (65/70) vs 59.5% (25/42), P<0.01], and the proportion of obstetric VTE risk assessment was significantly higher than that of the specialized hospital group [85.7% (60/70) vs 66.7% (28/42), P=0.017]. (2) Among the 88 hospitals using the VTE risk assessment sheet, the utilization rate of the evaluation sheet recommended by the obstetric VTE guideline in specialized hospitals (78.6%, 22/28) was higher than that of general hospitals (48.3%, 29/60), and the difference was statistically significant ( P=0.007). (3) The proportion of VTE incidence in general hospital group was 0.126% (360/286 517), and the proportion of VTE incidence in specialized hospital group was 0.032% (154/484 111). Logistic regression analysis showed that the OR value of the specialized hospital group was 0.253 (95% CI: 0.209-0.305) and the difference was statistically significant ( P<0.01) compared with the general hospital group. After adjusting the hospital level, B-ultrasonography and CTPA accessibility, whether to establish a risk assessment and obstetric VTE system, the proportion of VTE incidence in specialized hospitals was still lower than that in general hospitals ( OR=0.307, 95% CI: 0.251-0.376, P<0.01). Conclusions:General hospitals have higher access to resources for diagnosing VTE than specialized hospitals, and the VTE evaluation system is better implemented. The utilization rate of the obstetric VTE guidelines in specialized hospitals is higher than that of general hospitals.