1.Quality evaluation of Huocao based on UPLC fingerprint and multi-component content determination.
Zheng-Ming YANG ; Ci-Ga DIJIU ; Jian-Long LAN ; Jiang LUO ; Yue-Bu HAILAI ; Tao WANG ; Wen-Bing LI ; Ying LI ; Yuan LIU
China Journal of Chinese Materia Medica 2023;48(11):3000-3013
		                        		
		                        			
		                        			Huocao(a traditional Chinese herbal medicine) moxibustion is a characteristic technology in Yi medicine suitable for cold-dampness diseases. Huocao, as the moxibustion material, is confusedly used in clinical practice and little is known about its quality control. In this study, UPLC method was used to establish the chemical fingerprint of non-volatile components in Huocao, and the contents of eight phenolic acids such as chlorogenic acid were determined. Multivariate statistical analysis was performed to obtain the indicator components of Huocao for quality evaluation, and thus a comprehensive evaluation system for the quality of Huocao was built. The UPLC fingerprints of 49 batches of Huocao were established, and there were 20 common peaks, of which eight phenolic acids including neochlorogenic acid and chlorogenic acid were identified. Except for three batches of Huocao, the similarity of the other 46 batches was higher than 0.89, suggesting that the established fingerprint method could be used for quality control of the medicinal herb. The correlation coefficient between entropy weight score of the eight phenolic acids and comprehensive fingerprint score in Huocao was 0.875(P<0.01), which indicated that the eight phenolic acids could be used as indicator components for the quality evaluation of Huocao. Furthermore, in multivariate statistical analysis on the common peaks of fingerprint and the contents of the eight phenolic acids, chlorogenic acid, isochlorogenic acid A and isochlorogenic acid C were screened to be the indicator components. The results revealed that the proposed method achieved a simple and accurate quality control of Huocao based on UPLC fingerprint and multi-component content determination, which provided useful data for establishing the quality standard of Huocao.
		                        		
		                        		
		                        		
		                        			Chlorogenic Acid
		                        			;
		                        		
		                        			Entropy
		                        			;
		                        		
		                        			Hydroxybenzoates
		                        			;
		                        		
		                        			Quality Control
		                        			
		                        		
		                        	
2.Yinlai Decoction Protects Microstructure of Colon and Regulates Serum Level of D-Lactic Acid in Pneumonia Mice Fed with High-Calorie and High-Protein Diet.
Yun-Hui WANG ; He YU ; Tie-Gang LIU ; Teck Chuan KONG ; Zi-An ZHENG ; Yu-Xiang WAN ; Chen BAI ; Yu HAO ; Ying-Qiu MAO ; Jun WU ; Jing-Nan XU ; Li-Jun CUI ; Yu-Han WANG ; Yan-Ran SHAN ; Ying-Jun SHAO ; Xiao-Hong GU
Chinese journal of integrative medicine 2023;29(8):714-720
		                        		
		                        			OBJECTIVE:
		                        			To investigate the effect of Yinlai Decoction (YD) on the microstructure of colon, and activity of D-lactic acid (DLA) and diamine oxidase (DAO) in serum of pneumonia mice model fed with high-calorie and high-protein diet (HCD).
		                        		
		                        			METHODS:
		                        			Sixty male Kunming mice were randomly divided into 6 groups by the random number table method: normal control, pneumonia, HCD, HCD with pneumonia (HCD-P), YD (229.2 mg/mL), and dexamethasone (15.63 mg/mL) groups, with 10 in each group. HCD mice were fed with 52% milk solution by gavage. Pneumonia mice was modeled with lipopolysaccharide inhalation and was fed by gavage with either the corresponding therapeutic drugs or saline water, twice daily, for 3 days. After hematoxylin-eosin staining, the changes in the colon structure were observed under light microscopy and transmission electron microscope, respectively. Enzyme-linked immunosorbent assay was used to detect the protein levels of DLA and DAO in the serum of mice.
		                        		
		                        			RESULTS:
		                        			The colonic mucosal structure and ultrastructure of mice in the normal control group were clear and intact. The colonic mucosal goblet cells in the pneumonia group tended to increase, and the size of the microvilli varied. In the HCD-P group, the mucosal goblet cells showed a marked increase in size with increased secretory activity. Loose mucosal epithelial connections were also observed, as shown by widened intercellular gaps with short sparse microvilli. These pathological changes of intestinal mucosa were significantly reduced in mouse models with YD treatment, while there was no significant improvement after dexamethasone treatment. The serum DLA level was significantly higher in the pneumonia, HCD, and HCD-P groups as compared with the normal control group (P<0.05). Serum DLA was significantly lower in the YD group than HCD-P group (P<0.05). Moreover, serum DLA level significantly increased in the dexamethasone group as compared with the YD group (P<0.01). There was no statistical significance in the serum level of DAO among groups (P>0.05).
		                        		
		                        			CONCLUSIONS
		                        			YD can protect function of intestinal mucosa by improving the tissue morphology of intestinal mucosa and maintaining integrity of cell connections and microvilli structure, thereby reducing permeability of intestinal mucosa to regulate the serum levels of DLA in mice.
		                        		
		                        		
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Lactic Acid/pharmacology*
		                        			;
		                        		
		                        			Intestinal Mucosa
		                        			;
		                        		
		                        			Colon/pathology*
		                        			;
		                        		
		                        			Dexamethasone/pharmacology*
		                        			;
		                        		
		                        			Diet, High-Protein
		                        			;
		                        		
		                        			Pneumonia/pathology*
		                        			
		                        		
		                        	
3.Sodium butyrate activates HMGCS2 to promote ketone body production through SIRT5-mediated desuccinylation.
Yanhong XU ; Xiaotong YE ; Yang ZHOU ; Xinyu CAO ; Shiqiao PENG ; Yue PENG ; Xiaoying ZHANG ; Yili SUN ; Haowen JIANG ; Wenying HUANG ; Hongkai LIAN ; Jiajun YANG ; Jia LI ; Jianping YE
Frontiers of Medicine 2023;17(2):339-351
		                        		
		                        			
		                        			Ketone bodies have beneficial metabolic activities, and the induction of plasma ketone bodies is a health promotion strategy. Dietary supplementation of sodium butyrate (SB) is an effective approach in the induction of plasma ketone bodies. However, the cellular and molecular mechanisms are unknown. In this study, SB was found to enhance the catalytic activity of 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), a rate-limiting enzyme in ketogenesis, to promote ketone body production in hepatocytes. SB administrated by gavage or intraperitoneal injection significantly induced blood ß-hydroxybutyrate (BHB) in mice. BHB production was induced in the primary hepatocytes by SB. Protein succinylation was altered by SB in the liver tissues with down-regulation in 58 proteins and up-regulation in 26 proteins in the proteomics analysis. However, the alteration was mostly observed in mitochondrial proteins with 41% down- and 65% up-regulation, respectively. Succinylation status of HMGCS2 protein was altered by a reduction at two sites (K221 and K358) without a change in the protein level. The SB effect was significantly reduced by a SIRT5 inhibitor and in Sirt5-KO mice. The data suggests that SB activated HMGCS2 through SIRT5-mediated desuccinylation for ketone body production by the liver. The effect was not associated with an elevation in NAD+/NADH ratio according to our metabolomics analysis. The data provide a novel molecular mechanism for SB activity in the induction of ketone body production.
		                        		
		                        		
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Butyric Acid/metabolism*
		                        			;
		                        		
		                        			Ketone Bodies/metabolism*
		                        			;
		                        		
		                        			Liver/metabolism*
		                        			;
		                        		
		                        			Hydroxybutyrates/metabolism*
		                        			;
		                        		
		                        			Down-Regulation
		                        			;
		                        		
		                        			Sirtuins/metabolism*
		                        			;
		                        		
		                        			Hydroxymethylglutaryl-CoA Synthase/metabolism*
		                        			
		                        		
		                        	
4.Prognostic value of serum sodium variability within 72 hours and lactic acid combined with severity score in patients with sepsis.
Hang CHI ; Hongliang WANG ; Qingshu LI ; Zhiyuan LIAN ; Canxiu ZHANG ; Shuhu ZHANG ; Dan HU
Chinese Critical Care Medicine 2023;35(5):458-462
		                        		
		                        			OBJECTIVE:
		                        			To explore the predictive value of serum sodium variability within 72 hours, lactic acid (Lac), sequential organ failure assessment (SOFA) and acute physiology and chronic health evaluation II (APACHE II) in predicting the 28-day prognosis of sepsis patients.
		                        		
		                        			METHODS:
		                        			The clinical data of patients with sepsis admitted to the department of intensive care unit (ICU) of the Affiliated Qingdao Municipal Hospital of Qingdao University from December 2020 to December 2021 were retrospectively analyzed, including age, gender, previous medical history, temperature, heart rate, respiratory rate, systolic pressure, diastolic pressure, white blood cell count (WBC), hemoglobin (Hb), platelet count (PLT), C-reactive protein (CRP), pH value, arterial partial pressure of oxygen (PaO2), arterial partial pressure of carbon dioxide (PaCO2), Lac, prothrombin time (PT), activated partial thromboplastin time (APTT), serum creatinine (SCr), total bilirubin (TBil), albumin (Alb), SOFA, APACHE II score, and 28-day prognosis. Multivariate Logistic regression was used to analyze the risk factors of death in sepsis patients. Receiver operator characteristic curve (ROC curve) was used to analyze the predictive value of serum sodium variability within 72 hours, Lac, SOFA, APACHE II alone and in combination on the prognosis of patients with sepsis.
		                        		
		                        			RESULTS:
		                        			A total of 135 patients with sepsis were included, 73 survived and 62 died at 28 days, with 28-day mortality of 45.93%. (1) Compared with the survival group, SOFA, APACHE II, Lac and serum sodium variability within 72 hours in the death group were significantly higher [SOFA: 10.00 (8.00, 12.00) vs. 6.00 (5.00, 8.00), APACHE II: 18.00 (16.00, 21.25) vs. 13.00 (11.00, 15.00), Lac (mmol/L): 3.55 (2.90, 4.60) vs. 2.00 (1.30, 2.80), serum sodium variability within 72 hours: 3.4% (2.6%, 4.2%) vs. 1.4% (1.1%, 2.5%)], the differences were statistically significant (all P < 0.01). (2) Multivariate Logistic regression showed that SOFA, APACHE II, Lac, serum sodium variability within 72 hours were independent risk factors of prognosis in patients with sepsis [SOFA: odds ratio (OR) = 1.479, 95% confidence interval (95%CI) was 1.114-1.963, P = 0.007; APACHE II: OR = 1.163, 95%CI was 1.009-1.340, P = 0.037; Lac: OR = 1.387, 95%CI was 1.014-1.896, P = 0.040; serum sodium variability within 72 hours: OR = 1.634, 95%CI was 1.102-2.423, P = 0.015]. (3) ROC curve analysis showed that SOFA, APACHE II, Lac and serum sodium variability within 72 hours had certain predictive value for the prognosis of sepsis patients [SOFA: the area under the ROC curve (AUC) = 0.858, 95%CI was 0.795-0.920, P = 0.000; APACHE II: AUC = 0.845, 95%CI was 0.776-0.913, P = 0.000; Lac: AUC = 0.840, 95%CI was 0.770-0.909, P = 0.000; serum sodium variability within 72 hours: AUC = 0.842, 95%CI was 0.774-0.910, P = 0.000]. The combined predictive value of the four indicators (AUC = 0.917, 95%CI was 0.870-0.965, P = 0.000) was higher than that of any single indicator, and has higher specificity (79.5%) and sensitivity (93.5%), indicating that the combined index has higher predictive value for the prognosis of sepsis patients than any single index.
		                        		
		                        			CONCLUSIONS
		                        			SOFA, APACHE II, Lac, serum sodium variability within 72 hours are independent risk factors for 28-day death in patients with sepsis. The combination of SOFA score, APACHE II score, Lac and serum sodium variability within 72 hours has higher predictive value for prognosis than single index.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Lactic Acid
		                        			;
		                        		
		                        			Prognosis
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Sepsis
		                        			;
		                        		
		                        			Sodium
		                        			
		                        		
		                        	
5.Correlation between four limbs perfusion index and lactic acid in patients with severe neurological diseases.
Wen GUO ; Long MA ; Tuerxun TUERHONG ; Xiaopeng LI ; Bo LIU ; Zhiyi XIE ; Xiangyou YU
Chinese Critical Care Medicine 2023;35(5):509-512
		                        		
		                        			OBJECTIVE:
		                        			To observe the correlation between the four limbs perfusion index (PI) and blood lactic acid in patients with neurosis, and evaluate the predictive value of PI on microcirculation perfusion metabolic disorder in patients with neurosis.
		                        		
		                        			METHODS:
		                        			A prospective observational study was conducted. Adult patients admitted to the department of neurological intensive care unit (NICU) of the First Affiliated Hospital of Xinjiang Medical University from July 1 to August 20 in 2020 were enrolled. Under the condition of indoor temperature controlled at 25 centigrade, all patients were placed in the supine position, and the blood pressure, heart rate, PI of both fingers and thumb toes and arterial blood lactic acid were measured within 24 hours and 24-48 hours after NICU. The difference of four limbs PI at different time periods and its correlation with lactic acid were compared. Receiver operator characteristic curve (ROC curve) was used to evaluate the predictive value of four limbs PI on patients with microcirculatory perfusion metabolic disorder.
		                        		
		                        			RESULTS:
		                        			A total of 44 patients with neurosis were enrolled, including 28 males and 16 females; average age (61.2±16.5) years old. There were no significant differences in PI of the left index finger and the right index finger [2.57 (1.44, 4.79) vs. 2.70 (1.25, 5.33)], PI of the left toe and the right toe [2.09 (0.85, 4.76) vs. 1.88 (0.74, 4.32)] within 24 hours after entering the NICU, and the PI of the left index finger and the right index finger [3.17 (1.49, 5.07) vs. 3.14 (1.33, 5.36)], PI of the left toe and the right toe [2.07 (0.75, 5.20) vs. 2.07 (0.68, 4.67)] at 24-48 hours after NICU admission (all P > 0.05). However, compared to the PI of the upper and lower limbs on the same side, except for the 24-48 hours after ICU of the PI difference between the left index finger and the left toe (P > 0.05), the PI of the toe was lower than that of the index finger at the other time periods (all P < 0.05). The correlation analysis showed that the PI value of four limbs of patients in both time periods were significantly negatively correlated with arterial blood lactic acid (the r values of the left index finger, the right index finger, the left toe and the right toe were -0.549, -0.482, -0.392 and -0.343 respectively within 24 hours after entering the NICU; the r values of the left index finger, the right index finger, the left toe and the right toe were -0.331, -0.292, -0.402 and -0.442 respectively after entering the NICU 24-48 hours, all P < 0.05). Taking lactic acid ≥ 2 mmol/L as the diagnostic standard for metabolic disorder of microcirculation perfusion (total 27 times, accounting for 30.7%). The efficacy of four limbs PI in predicting microcirculation perfusion metabolic disorder were compared. ROC curve analysis showed that the area under the curve (AUC) and 95% confidence interval (95%CI) of left index finger, right index finger, left toe and right toe predicting microcirculation perfusion metabolic disorder were 0.729 (0.609-0.850), 0.767 (0.662-0.871), 0.722 (0.609-0.835), 0.718 (0.593-0.842), respectively. There was no significant difference in AUC compare with each other (all P > 0.05). The cut-off value of PI of right index finger for predicting microcirculation perfusion metabolic disorder was 2.46, the sensitivity was 70.4%, the specificity was 75.4%, the positive likelihood ratio was 2.86, and the negative likelihood ratio was 0.30.
		                        		
		                        			CONCLUSIONS
		                        			There are no significant differences in PI of bilateral index fingers, bilateral toes in patients with neurosis. However, unilateral upper and lower limbs showed lower PI in the toe than in the index finger. There is a significantly negatively correlation between PI and arterial blood lactic acid in all four limbs. PI can predict the metabolic disorder of microcirculation perfusion, and its cut-off value is 2.46.
		                        		
		                        		
		                        		
		                        			Adult
		                        			;
		                        		
		                        			Female
		                        			;
		                        		
		                        			Male
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Middle Aged
		                        			;
		                        		
		                        			Aged
		                        			;
		                        		
		                        			Lactic Acid
		                        			;
		                        		
		                        			Microcirculation
		                        			;
		                        		
		                        			Perfusion Index
		                        			;
		                        		
		                        			Lower Extremity
		                        			;
		                        		
		                        			Area Under Curve
		                        			;
		                        		
		                        			Nervous System Diseases
		                        			
		                        		
		                        	
6.Analysis of clinical characteristics and risk factors of early heat stroke-related acute liver injury.
Aiming LIU ; Zunguo PU ; Lulu CHU ; Hongsheng DING ; Yaqing ZHOU
Chinese Critical Care Medicine 2023;35(7):724-729
		                        		
		                        			OBJECTIVE:
		                        			To analyze the clinical characteristics and risk factors of early acute liver injury in patients with heat stroke (HS), and to provide basis for early identification of HS-related liver injury and its pathogenesis in clinical practice.
		                        		
		                        			METHODS:
		                        			The clinical data of patients with HS admitted to the department of critical care medicine of Haian People's Hospital from June 2015 to August 2022 were retrospectively analyzed. The patients with HS were divided into early liver injury group and early non-liver injury group according to the occurrence of acute liver injury within 24 hours of admission. The differences of basic data, clinical data, laboratory indexes and clinical outcomes of the two groups were analyzed. Logistic regression was used to analyze the risk factors for early HS-related acute liver injury, and receiver operator characteristic (ROC) curves were drawn to evaluate their value in predicting the occurrence of early HS-related acute liver injury.
		                        		
		                        			RESULTS:
		                        			A total of 76 patients with HS were enrolled, and 46 patients with acute liver injury, accounting for 60.53%. In the early liver injury group, 14 patients (30.43%) had elevated aminotransferase alone, 9 patients (19.57%) had elevated total bilirubin (TBil) alone, and 23 patients (50.00%) had elevated both aminotransferase and TBil. Among the patients with elevated aminotransferases, 24 patients (64.87%) had mild elevation, 5 patients (13.51%) had moderate elevation, 8 patients (21.62%) had severe elevation. Compared with the early non-liver injury group, acute physiology and chronic health evaluation II (APACHE II), sequential organ failure assessment (SOFA), arterial blood lactate (Lac), interleukin-6 (IL-6), procalcitonin (PCT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), TBil, γ-gamma glutamyl transferase (γ-GGT), lactate dehydrogenase (LDH), creatine kinase (CK), MB isoenzyme of creatine kinase (CK-MB), cardiac troponin I (cTnI), myoglobin (MYO), N-terminal B-type pro-brain natriuretic peptide (NT-proBNP), prothrombin time (PT), activated partial thromboplastin time (APTT), D-dimer in the early liver injury group were significantly increased, while platelet count (PLT) were significantly decreased within 24 hours after admission, the 28-day mortality was significantly increased [28.26% (13/46) vs. 6.67% (2/30)], and the differences were statistically significant (all P < 0.05). Univariate Logistic regression analysis showed that APACHE II score, SOFA score, PLT, Lac, IL-6, PCT, γ-GGT, LDH, CK, CK-MB, cTnI, MYO, PT, APTT, D-dimer were risk factors of early HS-related acute liver injury (all P < 0.05). Multivariate Logistic regression analysis showed that PLT, IL-6, and LDH were independent risk factors of early HS-related acute liver injury [odds ratio (OR) and 95% confidence interval (95%CI) were 0.986 (0.974-0.998), 1.027 (1.012-1.041), and 1.002 (1.000-1.004), all P < 0.05]. The ROC curve analysis showed that the area under the ROC curve (AUC) of PLT, IL-6 and LDH for predicting the occurrence of early HS-related acute liver injury was 0.672 (95%CI was 0.548-0.797), 0.897 (95%CI was 0.824-0.971) and 0.833 (95%CI was 0.739-0.927), respectively. IL-6 had the highest predictive value for early HS-related liver injury. When the optimal diagnostic threshold of IL-6 was 48.25 ng/L, the sensitivity was 95.7%, the specificity was 73.3%, and the predictive value of PLT was the lowest.
		                        		
		                        			CONCLUSIONS
		                        			The early HS-related liver injury is mainly manifested as the simultaneous elevation of aminotransferase and TBil, and most of cases are mild liver injury. PLT, IL-6 and LDH are independent risk factors of early HS-related acute liver injury.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Prognosis
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Interleukin-6
		                        			;
		                        		
		                        			ROC Curve
		                        			;
		                        		
		                        			Sepsis/diagnosis*
		                        			;
		                        		
		                        			Heat Stroke/complications*
		                        			;
		                        		
		                        			Risk Factors
		                        			;
		                        		
		                        			Alanine Transaminase
		                        			;
		                        		
		                        			Creatine Kinase, MB Form
		                        			;
		                        		
		                        			Lactic Acid
		                        			;
		                        		
		                        			Creatine Kinase
		                        			
		                        		
		                        	
7.Lactate-induced up-regulation of PLEKHA4 promotes proliferation and apoptosis of human glioma cells.
Jingjing YE ; Wenqin XU ; Bangsheng XI ; Nengqian WANG ; Tianbing CHEN
Journal of Southern Medical University 2023;43(7):1071-1080
		                        		
		                        			OBJECTIVE:
		                        			To investigate the effect of lactic acid-induced upregulation of PLEKHA4 expression on biological behaviors of glioma cells and the possible molecular mechanism.
		                        		
		                        			METHODS:
		                        			GEO database and GEPIA2 website were used to analyze the relationship between PLEKHA4 expression level and the pathological grade of glioma. A specific PLEKHA4 siRNA was transfected in glioma U251 and T98G cells, and the changes in cell proliferation ability were assessed by real-time cell analysis technology and Edu experiment. The colony-forming ability of the cells was evaluated using plate cloning assay, and cell cycle changes and cell apoptosis were analyzed with flow cytometry. The mRNA expression of PLEKHA4 was detected by PCR in glioma samples and controls and in glioma cells treated with lactic acid and glucose. Xenograft mice in vivo was used to detect tumor formation in nude mice; Western blotting was used to detect the expressions of cyclinD1, CDK2, Bcl2, β-catenin and phosphorylation of the key proteins in the MAPK signaling pathway.
		                        		
		                        			RESULTS:
		                        			The results of GEO database and online website analysis showed that PLEKHA4 was highly expressed in glioma tissues and was associated with poor prognosis; PLEKHA4 knockdown obviously inhibited the proliferation and attenuated the clone-forming ability of the glioma cells (P < 0.05). Flow cytometry showed that PLEKHA4 knockdown caused cell cycle arrest in G1 phase and promoted apoptosis of the cells (P < 0.01). PLEKHA4 gene mRNA expression was increased in glioma samples and glioma cells after lactate and glucose treatment (P < 0.01). PLEKHA4 knockdown, tumor formation ability of nude mice decreased; PLEKHA4 knockdown obviously lowered the expression of cyclinD1, CDK2, Bcl2 and other functional proteins, inhibited the phosphorylation of ERK and p38 and reduced the expression of β-catenin protein (P < 0.01).
		                        		
		                        			CONCLUSION
		                        			PLEKHA4 knockdown inhibited the proliferation of glioma cells and promoted apoptosis by inhibiting the activation of the MAPK signaling pathway and expression of β-catenin. Lactic acid produced by glycolysis upregulates the expression of PLEKHA4 in glioma cells.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Up-Regulation
		                        			;
		                        		
		                        			beta Catenin/metabolism*
		                        			;
		                        		
		                        			Mice, Nude
		                        			;
		                        		
		                        			Brain Neoplasms/pathology*
		                        			;
		                        		
		                        			Lactic Acid
		                        			;
		                        		
		                        			Cell Line, Tumor
		                        			;
		                        		
		                        			Glioma/pathology*
		                        			;
		                        		
		                        			Cell Proliferation
		                        			;
		                        		
		                        			Apoptosis
		                        			;
		                        		
		                        			Proto-Oncogene Proteins c-bcl-2/metabolism*
		                        			;
		                        		
		                        			RNA, Messenger/genetics*
		                        			;
		                        		
		                        			Gene Expression Regulation, Neoplastic
		                        			
		                        		
		                        	
8.Physiological regulation of salicylic acid on Helianthus tubeuosus upon copper stress and root FTIR analysis.
Jinxiang AI ; Jieke GE ; Ziyi ZHANG ; Wenqian CHEN ; Jiayi LIANG ; Xinyi WANG ; Qiaoyuan WU ; Jie YU ; Yitong YE ; Tianyi ZHOU ; Jinyi SU ; Wenwen LI ; Yuhuan WU ; Peng LIU
Chinese Journal of Biotechnology 2023;39(2):695-712
		                        		
		                        			
		                        			Phytoremediation plays an important role in the treatment of heavy metal pollution in soil. In order to elucidate the mechanism of salicylic acid (SA) on copper absorption, seedlings from Xuzhou (with strong Cu-tolerance) and Weifang Helianthus tuberosus cultivars (with weak Cu-tolerance) were selected for pot culture experiments. 1 mmol/L SA was sprayed upon 300 mg/kg soil copper stress, and the photosynthesis, leaf antioxidant system, several essential mineral nutrients and the changes of root upon copper stress were analyzed to explore the mechanism of copper resistance. The results showed that Pn, Tr, Gs and Ci upon copper stress decreased significantly compared to the control group. Meanwhile, chlorophyll a, chlorophyll b and carotenoid decreased with significant increase in initial fluorescence (F0), maximum photochemical quantum yield of PSⅡ (Fv/Fm), electron transfer rate (ETR) and photochemical quenching coefficient (qP) content all decreased. The ascorbic acid (AsA) content was decreased, the glutathione (GSH) value was increased, the superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activity in the leaves were decreased, and the peroxidase (POD) activity was significantly increased. SA increased the Cu content in the ground and root system, and weakened the nutrient uptake capacity of K, Ca, Mg, and Zn in the root stem and leaves. Spray of exogenous SA can maintain the opening of leaf stomata, improve the adverse effect of copper on photosynthetic pigment and PSⅡ reaction center. Mediating the SOD and APX activity started the AsA-GSH cycle process, effectively regulated the antioxidant enzyme system in chrysanthemum taro, significantly reduced the copper content of all parts of the plant, and improved the ion exchange capacity in the body. External SA increased the content of the negative electric group on the root by changing the proportion of components in the root, promoted the absorption of mineral nutrient elements and the accumulation of osmoregulatory substances, strengthened the fixation effect of the root on metal copper, and avoided its massive accumulation in the H. tuberosus body, so as to alleviate the inhibitory effect of copper on plant growth. The study revealed the physiological regulation of SA upon copper stress, and provided a theoretical basis for planting H. tuberosus to repair soil copper pollution.
		                        		
		                        		
		                        		
		                        			Antioxidants
		                        			;
		                        		
		                        			Copper
		                        			;
		                        		
		                        			Helianthus/metabolism*
		                        			;
		                        		
		                        			Salicylic Acid/pharmacology*
		                        			;
		                        		
		                        			Chlorophyll A/pharmacology*
		                        			;
		                        		
		                        			Spectroscopy, Fourier Transform Infrared
		                        			;
		                        		
		                        			Chlorophyll/pharmacology*
		                        			;
		                        		
		                        			Ascorbic Acid
		                        			;
		                        		
		                        			Superoxide Dismutase/metabolism*
		                        			;
		                        		
		                        			Photosynthesis
		                        			;
		                        		
		                        			Glutathione
		                        			;
		                        		
		                        			Plant Leaves
		                        			;
		                        		
		                        			Stress, Physiological
		                        			;
		                        		
		                        			Seedlings
		                        			
		                        		
		                        	
9.Immobilization of Lactobacillus bulgaricus with gellan gum and its application in continuous fermentation of D-lactic acid from corn straw.
Yongxin GUO ; Gang WANG ; Kexin LI ; Jiaqi HAN ; Huan CHEN ; Sitong ZHANG ; Yanli LI ; Guang CHEN
Chinese Journal of Biotechnology 2023;39(3):1083-1095
		                        		
		                        			
		                        			Biorefinery of chemicals from straw is an effective approach to alleviate the environmental pollution caused by straw burning. In this paper, we prepared gellan gum immobilized Lactobacillus bulgaricus T15 gel beads (LA-GAGR-T15 gel beads), characterized their properties, and established a continuous cell recycle fermentation process for D-lactate (D-LA) production using the LA-GAGR-T15 gel beads. The fracture stress of LA-GAGR-T15 gel beads was (91.68±0.11) kPa, which was 125.12% higher than that of the calcium alginate immobilized T15 gel beads (calcium alginate-T15 gel beads). This indicated that the strength of LA-GAGR-T15 gel beads was stronger, and the strain was less likely to leak out. The average D-LA production was (72.90±2.79) g/L after fermentation for ten recycles (720 h) using LA-GAGR-T15 gel beads as the starting strain and glucose as the substrate, which was 33.85% higher than that of calcium alginate-T15 gel beads and 37.70% higher than that of free T15. Subsequently, glucose was replaced by enzymatically hydrolyzed corn straw and fermented for ten recycles (240 h) using LA-GAGR-T15 gel beads. The yield of D-LA reached (1.74±0.79) g/(L·h), which was much higher than that of using free bacteria. The wear rate of gel beads was less than 5% after ten recycles, which indicated that LA-GAGR is a good carrier for cell immobilization and can be widely used in industrial fermentation. This study provides basic data for the industrial production of D-LA using cell-recycled fermentation, and provides a new way for the biorefinery of D-LA from corn straw.
		                        		
		                        		
		                        		
		                        			Fermentation
		                        			;
		                        		
		                        			Lactobacillus delbrueckii
		                        			;
		                        		
		                        			Zea mays
		                        			;
		                        		
		                        			Lactic Acid
		                        			;
		                        		
		                        			Alginates/chemistry*
		                        			;
		                        		
		                        			Glucose
		                        			
		                        		
		                        	
10.Effects of Huangqin Tang on NLRP3/Caspase-1 pathway in mice model of ulcerative colitis.
Meng-Ru LIU ; Hui LI ; Lan-Fu WEI ; Xiao-Tong LIU ; Zhen-Tao AN ; Li-Mei GU ; Yao-Zhou TIAN
China Journal of Chinese Materia Medica 2023;48(1):226-233
		                        		
		                        			
		                        			The aim of this study was to explore the effects of Huangqin Tang(HQT) on the NLRP3/Caspase-1 signaling pathway in mice with DSS-induced ulcerative colitis(UC). C57BL/6J mice were randomly divided into a blank group, a model group(DSS group), and low-, medium-and high-dose HQT groups(HQT-L, HQT-M, and HQT-H), and western medicine mesalazine group(western medicine group). The UC model was induced in mice. Subsequently, the mice in the HQT-L, HQT-M, HQT-H groups, and the western medicine group were given low-, medium-, high-dose HQT, and mesalazine suspension by gavage, respectively, while those in the blank and DSS groups were given an equal volume of distilled water by gavage. After 10 days of administration, the body weight, DAI scores, and colonic histopathological score of mice in each group were determined. The levels of IL-6, IL-10, IL-1β, and TNF-α in serum were determined by ELISA. The mRNA expression of NLRP3 and Caspase-1 in colon tissues was determined by RT-qPCR. The protein expression of NLRP3 and Caspase-1 in colon tissues was detected by immunohistochemistry. The results showed that compared with the blank group, the DSS group showed decreased body weight of mice and increased DAI scores and intestinal histopathological score. Compared with the DSS group, the HQT groups and the western medicine group showed improved DAI scores, especially in the HQT-M, HQT-H, and the western medicine groups(P<0.05). The intestinal histopathological scores of the HQT groups and the western medicine group significantly decreased, especially in the HQT-M, HQT-H, and the western medicine groups(P<0.05). In addition, compared with the blank group, the DSS group showed elevated expression of NLRP3 and Caspase-1 in colon tissues, increased serum levels of IL-6, IL-1β, and TNF-α, and decreased IL-10 level. Compared with the DSS group, the HQT groups and the western medicine group displayed decreased expression of NLRP3 and Caspase-1 in colon tissues, reduced serum levels of IL-6, IL-1β, and TNF-α, and increased IL-10 level. The improvement was the most significant in the HQT-H group and the western medicine group(P<0.01). In conclusion, HQT may reduce the expression of NLRP3 and Caspase-1 in colon tissues, reduce the se-rum levels of IL-6, IL-1β, and TNF-α, and increase the expression of IL-10 by regulating the classic pyroptosis pathway of NLRP3/Caspase-1, thereby improving the symptoms of intestinal injury and inflammatory infiltration of intestinal mucosa in DSS mice to achieve its therapeutic effect.
		                        		
		                        		
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Caspase 1/genetics*
		                        			;
		                        		
		                        			Colitis, Ulcerative/genetics*
		                        			;
		                        		
		                        			Colon
		                        			;
		                        		
		                        			Dextran Sulfate/adverse effects*
		                        			;
		                        		
		                        			Disease Models, Animal
		                        			;
		                        		
		                        			Interleukin-10/genetics*
		                        			;
		                        		
		                        			Interleukin-6/genetics*
		                        			;
		                        		
		                        			Mesalamine/pharmacology*
		                        			;
		                        		
		                        			Mice, Inbred C57BL
		                        			;
		                        		
		                        			NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
		                        			;
		                        		
		                        			Scutellaria baicalensis/chemistry*
		                        			;
		                        		
		                        			Tumor Necrosis Factor-alpha/metabolism*
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/pharmacology*
		                        			
		                        		
		                        	
            
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