Enhancing effect of macroporous adsorption resin on gamma-aminobutyric acid production by Enterococcus faecium in whole-cell biotransformation system.

Gamma-aminobutyric acid (GABA) biosynthesis depended to a great extent on the biotransformation characterization of glutamate decarboxylase (GAD) and process conditions. In this paper, the enhancing effect of D101 macroporous adsorption resin (MAR) on the GABA production was investigated based on the whole-cell biotransformation characterization of Enterococcus faecium and adsorption characteristics of D101 MAR. The results indicated that the optimal pH for reaction activity of whole-cell GAD and pure GAD was 4.4 and 5.0, respectively, and the pH range retained at least 50% of GAD activity was from 4.8 to 5.6 and 4.0-4.8, respectively. No substrate inhibition effect was observed on both pure GAD and whole-cell GAD, and the maximum activity could be obtained when the initial L-glutamic acid (L-Glu) concentration exceeded 57.6 mmol/L and 96.0 mmol/L, respectively. Besides, GABA could significantly inhibit the activity of whole-cell GAD rather than pure GAD. When the initial GABA concentration of the reaction solution remained 100 mmol/L, 33.51 ± 9.11% of the whole-cell GAD activity was inhibited. D101 MAR exhibited excellent properties in stabilizing the pH of the conversion reaction system, supplementing free L-Glu and removing excess GABA. Comparison of the biotransformation only in acetate buffer, the GABA production, with 50 g/100 mL of D101 MAR, was significantly increased by 138.71 ± 5.73%. D101 MAR with pre-adsorbed L-Glu could significantly enhance the production of GABA by gradual replenishment of free L-Glu, removing GABA and maintaining the pH of the reaction system, which would eventually make the GABA production more economical and eco-friendly.

Causal association and shared genetics between telomere length and COVID-19 outcomes: New evidence from the latest large-scale summary statistics.

Background: Observational studies suggested that leukocyte telomere length (LTL) is shortened in COVID-19 patients. However, the genetic association and causality remained unknown. Methods: Based on the genome-wide association of LTL (N = 472,174) and COVID-19 phenotypes (N = 1086,211-2597,856), LDSC and SUPERGNOVA were used to estimate the genetic correlation. Cross-trait GWAS meta-analysis, colocalization, fine-mapping analysis, and transcriptome-wide association study were conducted to explore the shared genetic etiology. Mendelian randomization (MR) was utilized to infer the causality. Upstream and downstream two-step MR was performed to investigate the potential mediating effects. Results: LDSC identified a significant genetic association between LTL and all COVID-19 phenotypes (rG < 0, p < 0.05). Six significant regions were observed for LTL and COVID-19 susceptibility and hospitalization, respectively. Colocalization analysis found rs144204502, rs34517439, and rs56255908 were shared causal variants between LTL and COVID-19 phenotypes. Numerous biological pathways associated with LTL and COVID-19 outcomes were identified, mainly involved in -immune-related pathways. MR showed that longer LTL was significantly associated with a lower risk of COVID-19 severity (OR [95% CI] = 0.81 [0.71-0.92], p = 1.24 ×10-3) and suggestively associated with lower risks of COVID-19 susceptibility (OR [95% CI] = 0.96 [0.92-1.00], p = 3.44 ×10-2) and COVID-19 hospitalization (OR [95% CI] = 0.89 [0.80-0.98], p = 1.89 ×10-2). LTL partially mediated the effects of BMI, smoking, and education on COVID-19 outcomes. Furthermore, six proteins partially mediated the causality of LTL on COVID-19 outcomes, including BNDF, QPCT, FAS, MPO, SFTPB, and APOF. Conclusions: Our findings suggested that shorter LTL was genetically associated with a higher risk of COVID-19 phenotypes, with shared genetic etiology and potential causality.

Comprehensive analysis of pyroptosis-related gene signatures for glioblastoma immune microenvironment and target therapy.

Glioblastoma (GBM) is a malignant brain tumour, but its subtypes (mesenchymal, classical, and proneural) show different prognoses. Pyroptosis is a programmed cell death relating to tumour progression, but its association with GBM is poorly understood. In this work, we collected 73 GBM samples (the Xiangya GBM cohort) and reported that pyroptosis involves tumour-microglia interaction and tumour response to interferon-gamma. GBM samples were grouped into different subtypes, cluster 1 and cluster 2, based on pyroptosis-related genes. Cluster 1 samples manifested a worse prognosis and had a more complicated immune landscape than cluster 2 samples. Single-cell RNA-seq data analysis supported that cluster 1 samples respond to interferon-gamma more actively. Moreover, the machine learning algorithm screened several potential compounds, including nutlin-3, for cluster 1 samples as a novel treatment. In vitro experiments supported that cluster 1 cell line, T98G, is more sensitive to nutlin-3 than cluster 2 cell line, LN229. Nutlin-3 can trigger oxidative stress by increasing DHCR24 expression. Moreover, pyroptosis-resistant genes were upregulated in LN229, which may participate against nutlin-3. Therefore, we hypothesis that GBM may be able to upregulate pyroptosis resistant related genes to against nutlin-3-triggered cell death. In summary, we conclude that pyroptosis highly associates with GBM progression, tumour immune landscape, and tumour response to nutlin-3.

Transcriptome Analysis in High Temperature Inhibiting Spermatogonial Stem Cell Differentiation In Vitro.

As one of the factors of male infertility, high temperature induces apoptosis of differentiated spermatogenic cells, sperm DNA oxidative damage, and changes in morphology and function of Sertoli cells. Spermatogonial stem cells (SSCs) are a type of germline stem cells that maintain spermatogenesis through self-renewal and differentiation. At present, however, the effect of high temperature on SSC differentiation remains unknown. In this study, an in vitro SSC differentiation model was used to investigate the effect of heat stress treatment on SSC differentiation, and RNA sequencing (RNA-seq) was used to enrich the key genes and pathways in high temperature inhibiting SSC differentiation. Results show that 2 days of 37 °C or 43 °C (30 min per day) heat stress treatment significantly inhibited SSC differentiation. The differentiation-related genes c-kit, stra8, Rec8, Sycp3, and Ovol1 were down-regulated after 2 and 4 days of heat stress at 37 °C. The transcriptome of SSCs was significantly differentially expressed on days 2 and 4 after heat stress treatment at 37 °C. In total, 1660 and 7252 differentially expressed genes (DEGs) were identified by RNA-seq in SSCs treated with heat stress at 37 °C for 2 and 4 days, respectively. KEGG pathway analysis showed that p53, ribosome, and carbon metabolism signaling pathways promoting stem cell differentiation were significantly enriched after heat stress treatment at 37 °C. In conclusion, 37 °C significantly inhibited SSC differentiation, and p53, ribosome, and carbon metabolism signaling pathways were involved in this differentiation inhibition process. The results of this study provide a reference for further investigation into the mechanism by which high temperature inhibits SSC differentiation.

Investigating the interaction between Shewanella oneidensis and phenazine 1-carboxylic acid in the microbial electrochemical processes.

Many exoelectrogens utilize small redox mediators for extracellular electron transfer (EET). Notable examples include Shewanella species, which synthesize flavins, and Pseudomonas species, which produce phenazines. In natural and engineered environments, redox-active metabolites from different organisms coexist. The interaction between Shewanella oneidensis and phenazine 1-carboxylic acid (PCA, a representative phenazine compound) was investigated to demonstrate exoelectrogens utilizing metabolites secreted by other organisms as redox mediators. After 24 h in a reactor with and without added PCA (1 µM), the anodic current generated by Shewanella was 235 ± 11 and 51.7 ± 2.8 µA, respectively. Shewanella produced oxidative current approximately three times as high with medium containing PCA as with medium containing the same concentration of riboflavin. PCA also stimulated inward EET in Shewanella. The strong effect of PCA on EET was attributed to its enrichment at the biofilm/electrode interface. The PCA voltammetric peak heights with a Shewanella bioanode were 25-30 times higher than under abiotic conditions. The electrochemical properties of PCA were also altered by the transition from two-electron to single-electron electrochemistry, which suggests PCA was bound between the electrode and cell surface redox proteins. This behavior would benefit electroactive bacteria, which usually dwell in open systems where mediators are present in low concentrations. Like flavins, PCA can be immobilized under both bioanode and biocathode conditions but not under metabolically inactive conditions. Shewanella rapidly transfers electrons to PCA via its Mtr pathway. Compared with wild-type Shewanella, the PCA reduction ability was decreased in gene knockout mutants lacking Mtr pathway cytochromes, especially in the mutants with severely undermined electrode-reduction capacities. These strains also lost the ability to immobilize PCA, even under current-generating conditions.

SYBR Green I-based real-time PCR targeting the rpoX gene for sensitive and rapid detection of Vibrio alginolyticus.

rpoX, a Vibrio alginolyticus specific stress regulating gene, was used to detect this fish pathogen by SYBR Green I-based real-time PCR. The specificity of the detection was confirmed in different samples. The minimum level of detection was 10(3) cells from pure culture and 10(2) cells from seawater.

Blue-light emitting aminated pectin for detecting Cu2+ ion.

This research studied the chemo-sensing of low-cost aminated pectin (PE) obtained by a facile calcination under ammonia gas at temperature no higher than 175 °C without excessive use of alkaline, acid or solvents. The ammonia gas was found to replace the hydroxyl and methoxyl group, enhancing the crystallinity and solubility of the resultant pectin than those calcined in air or in 5% H2. Though the increase of light absorption could be attributed mainly to the dehydration during calcination which caused the formation of CC double bond or aromatic ring, the N incorporation could be important to the photoluminescence (PL) emission. The PL quenching of the blue fluorescent aminated pectin showed a good linearity with the concentration of Cu2+, Fe3+ and the highest sensitivity toward Cu2+ among the investigated metal ions. In order to further increase the PL quenching toward Cu2+ and decrease the interference of Fe3+, a method involving H2O2 and ultraviolet illumination was developed to catalyze the oxidation of fluorophores on the polymer. This work provides new horizon on the modification and application of pectin in chemosensing.

Estimation of the biogeochemical reactivities of dissolved organic matter from modified biochars using color.

Modified biochar is widely used as a soil amendment in agricultural systems to improve crop yields and remove environmental pollutants. The water-soluble fraction of biochar, called biochar-derived dissolved organic matter (DOMBC), is the most active biochar component. However, the correlation between the optical properties of DOMBC and its biogeochemical activity remain unclear. In this study, one biochar and six modified derivatives were used to extract DOMBC and characterize its optical properties. The biogeochemical reactivities of DOMBC were determined using biodegradation, photodegradation, and electron-donating capacity assays. The results show that modification changes the biochar characteristics, leading to a variety of DOMBC properties. The DOMBC from modified biochars degrades more rapidly than the original biochar. On the other hand, modification reduces the redox functional groups in DOMBC, resulting in a lower electron-donating capacity of DOM samples. However, the modifications did not seem to affect photodegradation. Not all spectral parameters provide information about the correlations between the DOMBC properties and biogeochemical reactivity. However, two fundamental properties, that is, the specific UV absorbance at 254 nm (SUVA254, showing aromaticity) and spectral slopes over the ranges of 275-295 nm of the UV absorbance (S275-295, showing molecular weight), are the dominant factors affecting the biodegradation and electron-donating capacities of DOMBC. In this study, a rapid and straightforward method is presented, which can be used to characterize DOMBC and predict the reactivity of biochar that is used as an environmental amendment to minimize toxic organic compounds.

Role and mechanism of interleukin-8 in bone regeneration / 中国组织工程研究

BACKGROUND:Interleukin-8 is an important cytokine that has been found to play an important role in bone regeneration through multiple pathways. OBJECTIVE:To comprehensively review the action mechanism of interleukin-8 effects on bone regeneration to provide ideas for the following studies on interleukin-8. METHODS:By searching the China National Knowledge Infrastructure database for articles published from January 1999 to February 2023 and PubMed database for articles published from January 1985 to February 2023 reporting the role of interleukin-8 in bone-associated cells and vascularisation.Chinese and English search terms were"interleukin-8,bone repair,bone metabolism,mesenchymal stem cells,osteoblasts,osteoclasts,vascularization".The initial review yielded 508 articles in English and Chinese,and a total of 51 articles were included for review and analysis according to the inclusion and exclusion criteria. RESULTS AND CONCLUSION:According to the existing research,interleukin-8 can promote bone cell regeneration and assist bone healing through multiple pathways,which is mainly divided into three aspects:(1)Promote the proliferation and differentiation of bone cells such as mesenchymal stem cells and osteoblasts,and promote the development of cells in the direction of promoting bone healing;(2)interleukin-8 can promote angiogenesis and provide sufficient nutrition and oxygen for bone tissue,thus further improving the quality and stability of bone healing.(3)The appearance of interleukin-8 facilitates the expression of hypoxia-inducible factor-1α,vascular endothelial growth factor,and matrix metalloproteinase,which can create a microenvironment conducive to bone regeneration,thus promoting the regeneration and repair of bone tissue.In summary,interleukin-8 plays an important role in bone healing by promoting osteoblast proliferation and differentiation,facilitating angiogenesis and improving the mechanical properties of bone regeneration,as well as influencing bone metabolism through osteoclasts,mesenchymal stem cells,and other action sites.

Impairment of Autophagic Flux After Hypobaric Hypoxia Potentiates Oxidative Stress and Cognitive Function Disturbances in Mice / 神经科学通报·英文版

Acute hypobaric hypoxic brain damage is a potentially fatal high-altitude sickness. Autophagy plays a critical role in ischemic brain injury, but its role in hypobaric hypoxia (HH) remains unknown. Here we used an HH chamber to demonstrate that acute HH exposure impairs autophagic activity in both the early and late stages of the mouse brain, and is partially responsible for HH-induced oxidative stress, neuronal loss, and brain damage. The autophagic agonist rapamycin only promotes the initiation of autophagy. By proteome analysis, a screen showed that protein dynamin2 (DNM2) potentially regulates autophagic flux. Overexpression of DNM2 significantly increased the formation of autolysosomes, thus maintaining autophagic flux in combination with rapamycin. Furthermore, the enhancement of autophagic activity attenuated oxidative stress and neurological deficits after HH exposure. These results contribute to evidence supporting the conclusion that DNM2-mediated autophagic flux represents a new therapeutic target in HH-induced brain damage.

Protective effect of Dictyophora polysaccharide on neurotoxicity induced by sodium arsenite in rats / 中华地方病学杂志

Objective:To investigate the protective effect of Dictyophora polysaccharide on neurotoxicity induced by sodium arsenite in rats.Methods:Sixty SD rats (half males and half females) were selected and fed adaptively for one week. The rats were divided into a normal group ( n = 20, ordinary feed) and a modeling group [ n = 40, arsenic-containing feed (50 mg/kg sodium arsenite)] according to their body weight (80 - 100 g) by random number table method. After 12 weeks, the arsenic content in brain and blood of the rats ( n = 10) was measured to identify the arsenism model. After successful modeling, the rats in the modeling group were divided into Dictyophora polysaccharide group (arsenic-containing feed + 20 ml·kg -1·bw Dictyophora polysaccharide solution by gavage), and model group (arsenic-containing feed + equal volume distilled water by gavage), while the rats in the normal group (ordinary feed + equal volume distilled water by gavage) were retained, with 10 rats in each group for 4 weeks of intervention. Morris water maze test was used to assess the spatial learning and memory ability of the rats. Nissl staining was used to observe the pathological changes of the brain tissue, and the oxidative stress factors [superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA)], and inflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin -1β (IL-1β)] in the brain tissue of each group were measured. Results:Brain arsenic content of rats in the modeling group [(92.02 ± 13.37) μg/g] and blood arsenic content [(51.37 ± 19.33) μg/L] were higher than those of the normal group [(7.42 ± 3.21) μg/g and (2.74 ± 1.29) μg/L, t = - 6.91, - 6.06, P < 0.001]. The rat model of arsenic poisoning was successfully established. Compared with the normal group, the escape latency on the 1st, 3rd and 4th day and the first arrival time of rats in the model group were prolonged, the number of platform crossings was reduced, and the proportion of target quadrant residence time was decreased ( P < 0.05). Compared with the model group, the escape latency on the 4th day and the first arrival time of rats in the Dictyophora polysaccharide group were shortened, and the proportion of target quadrant residence time was prolonged ( P < 0.05). The results of Nissl staining showed that compared with the normal group, the number of Nissl bodies was decreased, the intercellular space increased, and the arrangement was disorderly in the model group; compared with the model group, the number of Nissl bodies was increased, most of the neurons were structurally intact. Compared with the normal group, the levels of SOD and GSH-Px in the brain tissue of rats in the model group were lower, and the levels of MDA, TNF-α and IL-1β were significantly higher ( P < 0.05). Compared with the model group, the levels of SOD and GSH-Px in the brain tissue of rats in the Dictyophora polysaccharide group were higher, while the levels of MDA, TNF-α and IL-1β were lower ( P < 0.05). In addition, the levels of SOD, GSH-Px, MDA and IL-1β in the Dictyophora polysaccharide group were not significantly different from those in the normal group ( P > 0.05). Conclusion:Diactyophora polysaccharide probably reduces the neurotoxicity damage caused by sodium arsenite in rats through antioxidant and antiinflammatory effects.

Expression of 5-methylcytosine and 5-hydroxymethylcytosine in bone tissue of rats with different types of skeletal fluorosis / 中华地方病学杂志

Objective:To learn about the levels of 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) in bone tissue of rats with different types of skeletal fluorosis and analyze their correlation.Methods:Thirty 4-week-old SPF grade healthy SD rats were selected. After adaptive feeding for 1 week, the rats were divided into control group (4 ml·kg -1·bw deionized water + standard maintenance diet), osteosclerosis group [20 mg·kg -1·bw sodium fluoride (NaF) + standard maintenance diet], and osteoporosis/osteomalacia group (20 mg·kg -1·bw NaF + low-calcium and low-protein partial diet) according to their body weight (100 - 120 g) by random number table method, with 10 rats in each group, half male and half female; gavaged 6 days each week and the experimental period was 5 months. At the end of the experiment, samples of rat heart blood and lower limb femur were collected. The contents of serum methyl donor S-adenosylmethionine (SAM) and its metabolite S-adenosylhomocysteine (SAH) in serum, and the levels of 5-mC and 5-hmC in bone tissue were measured by enzyme-linked immunosorbent assay (ELISA). Western blot was used to determine the expression of DNA methyltransferase (DNMTs) and DNA hydroxymethylase (TETs) in bone tissue of rats. The correlation between serum SAM content, SAM/SAH ratio and bone tissue 5-mC level, and between the bone tissue 5-mC level and 5-hmC level was analyzed. Results:Serum SAM [11.03 (7.06, 18.63), 3.96 (2.32, 9.09), 3.91 (2.35, 4.46) nmol/L], SAH content [(4.69 ± 0.55), (5.41 ± 1.13), (13.90 ± 1.09) ng/L], SAM/SAH ratio [2.58 (1.54, 4.12), 0.62 (0.52, 1.69), 0.14 (0.13, 0.15)] and bone tissue 5-mC [103.39 (97.37, 109.35), 52.50 (50.19, 68.13), 55.03 (49.97, 59.57) ng/L], 5-hmC levels [(32.61 ± 8.84), (56.96 ± 8.48), (20.34 ± 6.22) ng/L] in the control group, osteosclerosis group and osteoporosis/osteomalacia group were compared, and the differences were statistically significant beween three groups ( H/ F = 12.81, 284.24, 21.85, 19.37, 55.23, P < 0.01). Compared with the control group, the content of SAM, the ratio of SAM/SAH, the level of 5-mC in the osteosclerosis group and osteoporosis/osteomalacia group, and the level of 5-hmC in the osteoporosis/osteomalacia group were lower ( P < 0.05), while the content of SAH in the osteoporosis/osteomalacia group and the level of 5-hmC in the osteosclerosis group were higher ( P < 0.05). Compared with the osteosclerosis group, the content of SAH in the osteoporosis/osteomalacia group was higher, while the ratio of SAM/SAH and the level of 5-hmC were lower ( P < 0.05). Western blot showed that there were statistically significant differences in the expression levels of DNMT1, DNMT3A, DNMT3B, TET1 and TET2 protein in bone tissue of rats in the control group, osteosclerosis group, and osteoporosis/osteomalacia group ( F = 285.45, 345.58, 239.83, 311.52, 318.24, P < 0.001). Among them, the expression levels of DNMT1, DNMT3A and DNMT3B protein in the osteosclerosis group and osteoporosis/osteomalacia group were lower than those in the control group, and the expression levels of DNMT1, DNMT3A and DNMT3B protein in the osteosclerosis/osteomalacia group were lower than those in the osteosclerosis group ( P < 0.05); the expression levels of TET1 and TET2 protein in osteosclerosis group were higher than those in the control group and osteoporosis/osteomalacia group, and the expression levels of TET1 and TET2 protein in the osteoporosis/osteomalacia group were lower than those in the control group ( P < 0.05). The results of Spearman rank correlation analysis showed that the content of SAM and the ratio of SAM/SAH in the control group, osteosclerosis group and osteoporosis/osteomalacia group were positively correlated with the level of 5-mC in bone tissue ( rs = 0.89, 0.92, 0.81, 0.73, 0.87, 0.73, P < 0.05). The levels of 5-mC and 5-hmC in bone tissue of rats in each group were negatively correlated ( rs = - 0.69, - 0.68, - 0.72, P < 0.05). Conclusions:The level of 5-mC in bone tissue of osteosclerotic fluorosis rats is low, and the level of 5-hmC is high, while those of osteoporosis/osteomalacia fluorosis rats are lower. The difference of 5-mC level in bone tissue of rats with different types of skeletal fluorosis is not significant, which may be related to the difference of 5-hmC level in bone tissue.

Discussion on the staged treatment of chronic heart failure based on the theory of "deficiency, blood stasis, water and toxin" / 国际中医中药杂志

Deficiency, stasis, water and toxin are of great significance in the pathogenesis and pathologic evolution of chronic heart failure (CHF). Based on "deficiency, blood stasis, water and toxin", the pathogenesis and treatment of CHF were discussed in this article. It was found that in the pathogenesis, deficiency--deficiency of heart qi and deficiency of heart yang were the origin of the disease, and blood stasis, water and toxin were the markers of the disease. Among them, blood stasis was the central pathological link, and also an important mechanism that could aggravate the disease and cause a vicious cycle; water-phlegm and water dampness were the basic pathological products; toxin-heat toxin, water toxin, and stasis toxin were the final results of disease progress and product accumulation. In terms of treatment, CHF can be divided into four stages: early, middle, late and end. In the early stage, tonifying qi and regulating heart can be used for the treatment of root cause, and promoting blood circulation and water can be used for the treatment of symptoms; tonifying qi and yin and reinforcing the healthy qi, reducing blood stasis, purging turbid, and eliminating pathogenic factors can be used in the middle stage; reducing blood stasis and removing toxic materials should be used in the late stage, supplemented with warming yang and increasing urine excretion; astringing yang,generating body fluids, tonifying qi and yang should be used in the end stage. At the same time of treating by stages, attention should be paid to adhering to a holistic concept and dialectical treatment; pay attention to timing and flexible medication; adopting a combination of Chinese and Western approaches and integrating them.

Analysis on risk factors for prognosis of traumatic brain injury in adults and establishment of the prediction model / 中华创伤杂志

Objective:To analyze risk factors for prognosis of adult patients with traumatic brain injury (TBI), construct the prognostic model of TBI and evaluate its predictive value.Methods:A case-control study was used to analyze the clinical data of 522 patients with TBI admitted to Xijing Hospital of Air Force Medical University from March 2011 to September 2019, including 438 males and 84 females; aged 18-75 years [(44.9±15.0)years]. According to the Glasgow outcome score (GOS) at discharge, the patients were divided into good prognosis group (GOS 4-5 points, n=165) and poor prognosis group (GOS 1-3 points, n=357). The two groups were compared with regards to qualitative data such as sex, underlying diseases, causes of injury, multiple injuries, open injuries, intracranial foreign bodies, cerebral herniation, consciousness status on admission and at discharge, surgery, lung infection on admission, tracheostomy, ventilator-assisted ventilation, hospital-acquired pneumonia/pathogenic bacteria and intracranial infection, and quantitative data such as Glasgow coma score (GCS) on admission and at discharge, age, measurements on admission [systolic blood pressure, diastolic blood pressure, mean arterial pressure, temperature, heart rate, creatinine, urea nitrogen, blood sodium, blood potassium, blood glucose, prothrombin time (PT), activated partial thromboplastin time (APTT), platelets, international normalized ratio (INR), pupil size of both eyes] and length of hospital stay. Univariate analysis and Lasso regression analysis were used to screen the risk factors affecting the prognosis of TBI patients, and the selected influencing factors were included in multivariate Logistic regression analysis to identify independent risk factors and construct regression equations. R was used to draw a visual nomogram based on regression equation for predicting the prognosis of TBI patients. The prognostic predictive value of the nomogram was evaluated by using the receiver operating characteristic (ROC) curve, and the area under the curve (AUC), Youden index, sensitivity, specificity and consistency index (C index) were calculated. Results:Univariate analysis showed that there were significant differences between the two groups in underlying diseases, open injuries, cerebral herniation, consciousness status on admission and at discharge, lung infection on admission, tracheostomy, ventilator-assisted ventilation, hospital-acquired pneumonia/pathogenic bacteria, GCS on admission and at discharge, age, and measurements on admission (systolic blood pressure, mean arterial pressure, body temperature, heart rate, creatinine, urea nitrogen, blood potassium, blood glucose, PT, INR, pupil size of right eye) (all P<0.05 or 0.01). There were no significant differences between the two groups in gender, causes of injury, multiple injuries, intracranial foreign bodies, surgery, intracranial infection, measurements on admission (diastolic blood pressure, blood sodium, APTT, platelets, pupil size of left eye) and length of hospital stay (all P>0.05). After screening by Lasso regression model, the results of multivariate Logistic regression analysis showed that GCS on admission ( OR=0.67, 95% CI 0.62, 0.73, P<0.01), age ( OR=1.03, 95% CI 1.01, 1.04, P<0.01), blood glucose on admission ( OR=1.17, 95% CI 1.06, 1.30, P<0.01) and INR on admission ( OR=17.08, 95% CI 2.12, 137.89, P<0.01) could be used as the main risk factors to construct the prediction model, and the regression equation was constructed: Logit [ P/(1- P)]=-0.398× "GCS on admission"+0.024× "age"+0.158×"blood glucose on admission"+2.838×"INR on admission"-1.693. The AUC for the prognosis prediction in adult patients with TBI using R based on a visual nomogram model was 0.87 (95% CI 0.83, 0.89, P<0.01). The Youden index for the predicted probability was 0.60 (sensitivity of 85.2% and specificity of 75.2%), with the C index of 0.87. Conclusion:Age, GCS on admission, blood glucose on admission and INR on admission are the main risk factors affecting the prognosis of TBI in adults, and the nomogram drawn by these parameters can better predict their clinical outcome.

Unventilated indoor coal-fired stoves in Guizhou province, China: cellular and genetic damage in villagers exposed to arsenic in food and air.

BACKGROUND: Inorganic arsenic (iAs) is a well-known human carcinogen recognized by the World Health Organization and the International Agency for Research on Cancer. Currently, most iAs studies in populations are concerned with drinking water and occupational arsenicosis. In Guizhou province, arsenicosis caused by the burning of coal in unventilated indoor stoves is an unusual type of exposure. Because the poisoning mechanism involved in arsenicosis is as yet unknown and no effective therapy exists, progress has been slow on the prevention and therapy of arsenicosis. OBJECTIVES: We examined the relationship between arsenic (As) exposure from the burning of coal in unventilated indoor stoves and genetic damage in humans, using cellular and molecular indices. We selected villagers from Jiaole township, Guizhou province, China, who had been exposed to milligram levels of As daily via food and air contaminated by the burning of As-containing coal in unventilated indoor stoves. RESULTS: The As-exposed subjects from Jiaole were divided into four groups according to skin lesion symptoms: nonpatients, mild, intermediate, and severe arsenicosis. Another 53 villagers from a town 12 km from Jiaole were recruited as the external control group. In the four groups of exposed subjects, As concentrations in urine and hair were 76-145 microg/L and 5.4-7.9 microg/g, respectively. These values were higher than those in the external control group, which had As concentrations of 46 microg/L for urine and 1.6 microg/g for hair. We measured sister chromatid exchange and chromosomal aberrations to determine human chromosome damage, and for DNA damage, we measured DNA single-strand breaks and DNA-protein cross-links. All measurements were higher in the four exposed groups compared with the external control group. DNA repair was impaired by As exposure, as indicated by the mRNA of O-6-methylguanine-DNA methyltransferase (MGMT), X-ray repair complementing defective repair in Chinese hamster cells 1 (XRCC1), and, to a lesser extent, by the mismatch repair gene hMSH2 mRNA. The expression of mutant-type p53 increased with aggravation of arsenicosis symptoms, whereas the expression of p16-INK4(p16) decreased. p53 mutated at a frequency of 30-17% in the carcinoma (n = 10) and precarcinoma (n = 12) groups. No mutation was found in p16, although deletion was evident. Deletion rates were 8.7% (n = 23) and 38.9% (n = 18) in noncarcinoma and carcinoma groups, respectively. CONCLUSIONS: The results showed that long-term As exposure may be associated with damage of chromosomes and DNA, gene mutations, gene deletions, and alterations of DNA synthesis and repair ability.

Diverse coagulopathies in a rabbit model with different abdominal injuries.

BACKGROUND: Although coagulopathy can be very common in severe traumatic shock patients, the exact incidence and mechanism remain unclear. In this study, a traumatic shock rabbit model with special abdomen injuries was developed and evaluated by examining indicators of clotting and fibrinolysis. METHODS: Forty New Zealand white rabbits were randomly divided into four groups: group 1 (sham), group 2 (hemorrhage), group 3 (hemorrhage-liver injury), and group 4 (hemorrhage-liver injury/intestinal injury-peritonitis). Coagulation was detected by thromboelastography before trauma (T0), at 1 hour (T1) and 4 hours (T2) after trauma. RESULTS: Rabbits that suffered from hemorrhage alone did not differ in coagulation capacity compared with the sham group. The clot initiations (R times) of group 3 at T1 and T2 were both shorter than those of groups 1, 2, and 4 (P<0.05). In group 4, clot strength was decreased at T1 and T2 compared with those in groups 1, 2, and 3 (P<0.05), whereas the R time and clot polymerization were increased at T2 (P<0.05). The clotting angle significantly decreased in group 4 compared with groups 2 and 3 at T2 (P<0.05). CONCLUSION: This study suggests that different abdominal traumatic shock show diverse coagulopathy in the early phase. Isolated hemorrhagic shock shows no obvious effect on coagulation. In contrast, blunt hepatic injury with hemorrhage shows hypercoagulability, whereas blunt hepatic injury with hemorrhage coupled with peritonitis caused by a ruptured intestine shows a tendency toward hypocoagulability.

Effects of ginkgo biloba tablets on liver injury in patients with coal-burning-borne arsenism based on DNA damage and repair inhibition / 中华地方病学杂志

Objective:To investigate the role of DNA damage and repair inhibition in the effect of ginkgo biloba on liver injury in patients with coal-burning-borne arsenism.Methods:In March 2017, the investigation was conducted in Jiaole village arsenic poisoning area in Yuzhang Town, Xingren County, Guizhou Province. According to the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015) and the "Diagnostic Criteria of Occupational Toxic Hepatopathy" (GBZ 59-2010), 52 patients with arsenism were selected as the ginkgo biloba intervention group, and 49 cases of arsenism patients as intervention control group. Ginkgo biloba tablets were given orally for 3 months (1 tablet/time, 3 times/d) according to the commonly used clinical methods, and no other drugs were given to all subjects during the intervention period. The intervention control group was given placebo in the same way as that of ginkgo biloba intervention group. A total of 41 residents who did not burn high arsenic coal 12 km away with no abnormal liver function were selected as normal control group. Physical examinations were performed before the intervention and at the end of the intervention at 3 months. After receiving signed informed consent, morning urine and peripheral venous blood samples were collected to detect urinary arsenic content by inductively coupled plasma mass spectrometry (ICP-MS); liver function biochemical indexes [albumin (ALB), albumin/globulin (A/G), cholinesterase (CHE), total bile acid (TBA)] were determined by automatic biochemical analyzer, DNA damage by single-cell gel electrophoresis assay, and the expression of miR-145 (repair inhibition index) by qRT-PCR.Results:There were 116 subjects, 41 in normal control group, 39 in ginkgo biloba intervention group and 36 in intervention control group. In ginkgo biloba and intervention and intervention control groups, there was no significant difference in age, gender, smoking habits and drinking compared with normal control group ( P > 0.05). Urinary arsenic content, TBA level, DNA damage degree [comet tail DNA percentage (TailDNA%) and olive tail moment (OTM)] and plasma miR-145 expression level [(38.75 ± 19.09) μg/g Cr, (11.13 ± 1.55) μmol/L, 8.50 ± 0.88, 7.43 ± 0.68, 5.78 ± 0.75, respectively] in ginkgo biloba intervention group patients before intervention were higher than those in normal control group [(11.62 ± 5.33) μg/g Cr, (5.36 ± 0.87) μmol/L, 5.24 ± 0.33, 4.71 ± 0.29, 2.05 ± 0.27, respectively], the differences were statistically significant ( P < 0.05); the levels of ALB, A/G and CHE were significantly lower than those in normal control group ( P < 0.05). After the intervention of ginkgo biloba, urinary arsenic content, TBA level, DNA damage degree (TailDNA% and OTM) and plasma miR-145 expression level in patients were significantly lower than those before the intervention ( P < 0.05); the levels of ALB, A/G and CHE were significantly higher than those before the intervention ( P < 0.05). There was no significant difference in the above indexes before and after intervention in the intervention control group ( P > 0.05). The results of correlation analysis between DNA damage degree, miR-145 and liver function indexes after the intervention of ginkgo biloba showed that, DNA damage degree (TailDNA% and OTM) was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.34, - 0.33, - 0.48, - 0.31, - 0.31, - 0.42, P < 0.05), and positively correlated with the level of TBA ( r = 0.49, 0.48, P < 0.05); miR-145 was negatively correlated with the levels of ALB, A/G and CHE ( r = - 0.26, - 0.23, - 0.38, P < 0.05), which was positively correlated with the level of TBA ( r = 0.32, P < 0.05); and DNA damage degree was positively correlated with the expression of miR-145 ( r = 0.65, 0.52, P < 0.05). Conclusion:Ginkgo biloba tablets can alleviate the liver damage caused by arsenic through coal burning, and the mechanism of this process is related to its inhibition of miR-145 expression and reduction of DNA damage.

Micro RNA-373-3p is involved in regulation of autophagy and sunitinib sensitivity of glioblastoma cells via targeting autophagy-related gene 7 / 中华神经医学杂志

Objective:To investigate the influence and mechanism of micro RNA (miR)-373-3p in autophagy and sunitinib sensitivity of glioblastoma cells.Methods:U251 cells were routinely cultured in vitro; and some U251 cells were subjected to 50 μmol/L sunitinib treatment for 72 h to construct sunitinib-resistant U251 cell line. (1) Real-time reverse transcription quantitative PCR (RT-qPCR) was used to detect the miR-373-3p expression in U251 and sunitinib-resistant U251 cells. Sunitinib-resistant U251 cells were divided into blank control group, nonsense sequence group and miR-373-3p mimic group; cells in the latter 2 groups were transfected with nonsense sequence and miRNA-337-3p mimic, respectively; miR-373-3p expression was detected by RT-qPCR. Cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, and sunitinib-resistant U251+miR-373-3p mimic group; after each transfection, CCK-8 assay was used to evaluate the cell viability; TUNEL was used to detect the apoptotic rate; immunofluorescent assay was used to detect the expression of microtubule-associated protein light chain 3 (LC3); Western blotting was used to detect the expressions of apoptosis- and autophagy-associated proteins. (2) The pGL3-autophagy-related gene 7 (ATG7) wild-type (WT) and pGL3-ATG7 mutant type (MUT) plasmids were established; dual-luciferase reporter system was used to detect the cell luciferase activity in the miR-373-3p mimic group and nonsense sequence group. Cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, and sunitinib-resistant U251+miR-373-3p mimic group; after each transfection, RT-qPCR and Western blotting were used to detect the mRNA and protein expressions of ATG7 in the cells. (3) The sunitinib-resistant U251 cells were divided into blank control group, ATG7 negative control group, and ATG7 overexpression group; after each transfection, RT-qPCR and Western blotting were used to detect the ATG7 mRNA and protein expressions. U251 and sunitinib-resistant U251 cells were divided into U251 group, sunitinib-resistant U251 group, sunitinib-resistant U251+nonsense sequence group, sunitinib-resistant U251+miR-373-3p mimic group, sunitinib-resistant U251+miR-373-3p mimic+ATG7 negative control group, and sunitinib-resistant U251+miR-373-3p mimic+ATG7 overexpression group; after each transfection, CCK-8 assay was used to evaluate the cell apoptosis, TUNEL was used to examine the apoptotic rate, and Western blotting was employed to detect the expressions of apoptosis- and autophagy-associated proteins. Results:(1) As compared with that in the U251 cells, miR-373-3p was lowly expressed in sunitinib-resistant U251 cells, with statistic difference ( P<0.05). As compared with that in the blank control group and nonsense sequence group, miR-373-3p expression was significantly elevated in the miR-373-3p mimic group ( P<0.05). As compared with the U251 group, the sunitinib-resistant U251 group had significantly increased cell viability, significantly decreased cell apoptotic rate, statistically increased B lymphocytoma-2 (Bcl-2) and Beclin 1 protein expressions, and significantly increased LC3II/LC3I values, significantly decreased Bcl-2 associated X protein (Bax) and p62 protein expressions and cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had significantly decreased cell viability, significantly increased cell apoptotic rate, statistically decreased Bcl-2 and Beclin 1 protein expressions, and significantly decreased LC3II/LC3I values, significantly increased Bax and p62 protein expressions and cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the U251 group, the sunitinib-resistant U251 group had increased number of fluorescent particles labeled with LC3 and enhanced fluorescent intensity; as compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had decreased number of fluorescent particles labeled with LC3 and reduced fluorescent intensity. (2) The luciferase activity of pGL3-ATG7 WT plasmids in the miR-373-3p mimic group was signficantly lower than that in nonsense sequence group ( P<0.05). As compared with those in the U251 group, ATG7 mRNA and protein expressions were both significantly increased in the sunitinib-resistant U251 group ( P<0.05); as compared with those in the sunitinib-resistant U251+nonsense sequence group, ATG7 mRNA and protein expressions were both significantly decreased in the sunitinib-resistant U251+miR-373-3p mimic group ( P<0.05). (3) As compared with the blank control group and ATG7 negative control group, the ATG7 overexpression group had significantly increased ATG7 mRNA and protein expressions ( P<0.05). As compared with the sunitinib-resistant U251+nonsense sequence group, the sunitinib-resistant U251+miR-373-3p mimic group had significantly decreased cell viability, significantly increased cell apoptotic rate, statistically decreased Bcl-2 and Beclin 1 protein expressions, significantly decreased LC3II/LC3I values, significantly increased Bax and p62 protein expressions, and significantly increased cleaved Caspase3/Caspase 3 values ( P<0.05). As compared with the sunitinib-resistant U251+miR-373-3p mimic+ATG7 negative control group, the sunitinib-resistant U251+miR-373-3p mimic+ATG7 overexpression group had significantly increased cell viability, significantly decreased apoptotic rate, statistically increased Bcl-2 and Beclin 1 protein expressions, significantly increased LC3II/LC3I values, significantly decreased Bax and p62 protein expressions, and significantly decreased cleaved Caspase3/Caspase 3 values ( P<0.05) Conclusion:MiR-373-3p can enhance sunitinib sensitivity by regulating autophagy in glioblastoma cells, whose mechanism might be related to targeting ATG7.

Application value of right minimal invasive three-port technique of laparoscopic sleeve gastrec-tomy for the treatment of obesity / 中华消化外科杂志

Objective:To investigate the application value of right minimal invasive three-port technique of laparoscopic sleeve gastrectomy (RMIT-LSG) for the treatment of obesity.Methods:The retrospective and descriptive study was conducted. The clinical data of 66 obesity patients who underwent RMIT-LSG in the Sir Run Run Shaw Hospital of Zhejiang University School of Medicine from January to October 2021 were collected. There were 15 males and 51 females, aged 28.5(range, 16.0?54.0)years. The body mass index (BMI) of the 66 patients was (36.9±4.3)kg/m 2. There were 20 of the 66 patients combined with type 2 diabetes. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination or the WeChat to detect postoperative recovery of patients including body mass changing, BMI and complications 6 months after operation. The follow-up was up to December 2021. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical situations. All the 66 patients underwent RMIT-LSG successfully, without conversion to laparotomy or changing surgical method. The operation time and the volume of intraoperative blood loss of the 66 patients were (132±22)minutes and (14±8)mL, respectively. (2) Postoperative situations. The time to postoperative initial out-of-bed activities, time to postoperative first flatus, time to postoperative initial water intake, time to postoperative initial liquid food intake and duration of postoperative hospital stay of the 66 patients were (15±6)hours, (1.80±0.60)days, (1.00±0.20)days, (2.00±0.20)days and (3.40±0.60)days, respectively. Of the 66 patients, one case underwent post-operative abdominal hemorrhage at postoperative day 1 and received a second surgery for hemostasis. The patient with postoperative abdominal hemorrhage and other 65 patients recovered well without gastroparesis, gastric fistula, abdominal infection and other complication. (3) Follow-up. All the 66 patients were followed up for 6(range, 1?11)months. All the 66 patients completed the postoperative scar photography at postoperative 1 month, and results of scar photography showed concealed scar with good cosmetic effects. Twenty-seven of the 66 patients were followed up for 6 months after operation, with the weight loss, percentage of weight loss and decrease of BMI were (42±7)kg, 34.8%±2.9%, (14.2±1.9)kg/m 2, respectively. None of the 66 patient had innutrition during the follow-up. Conclusion:The RMIT-LSG is safe and feasible for the treatment of obesity, with a good cosmetic effect of the wound.

Application of the Mathieu combined tunnel technique for repairing glans dehiscence after failed hypospadias repair / 亚洲男科学杂志(英文版)

Repairing glans dehiscence after failed hypospadias repair is challenging for pediatric surgeons. Here, we introduced and evaluated a newly modified Mathieu technique, Mathieu combined tunnel (MCT), which involves multiple custom-designed flaps for the shortage of flap source material after repeated operations; we also constructed a tunnel to avoid the glans incision that may carry new risks of dehiscence. This retrospective study included 26 patients who were consecutively admitted to the First Affiliated Hospital of Sun Yat-Sen University (Guangzhou, China) for glans dehiscence repair after failed hypospadias repair from October 2014 to October 2020; sixteen patients underwent surgery using the MCT (MCT group) and ten patients underwent surgery using the tubularized incised plate (TIP) technique (TIP group). The operative time, blood loss, postoperative complications, normal urethral meatus rate, success rate, and Hypospadias Objective Penile Evaluation (HOPE) score were compared between the two groups. The MCT group achieved an overall satisfactory penile appearance and voiding function, with a higher rate of normal urethral meatus (15/16, 93.8%) and a lower rate of glans dehiscence (1/16, 6.2%), compared with the TIP group (70.0% and 30.0%, respectively). However, these differences were not statistically significant, possibly because of the limited number of patients (all P > 0.05). Mean postoperative HOPE scores were similar in the MCT group (mean ± standard deviation: 8.83 ± 0. 89) and TIP group (8.94 ± 0.57) (P > 0.05). No significant differences were found between the two groups in terms of blood loss and success rate, nor in the rates of various complications (e.g., fistula, urethral stricture, and glans dehiscence). In conclusion, the MCT technique appears to be feasible and reliable for repairing glans dehiscence after failed hypospadias repair.