1.Effect of high-dose vitamin B6 on stress-induced liver cell death in rats with severe trauma and the possible mechanism
Yinjie ZHANG ; Zhihuai WANG ; Xuelin TANG ; Haiyang ZHOU ; Peng GAO ; Chunfu ZHU ; Zhongzhi JIA ; Maoxing YUE ; Xihu QIN
Chinese Journal of Trauma 2024;40(6):558-568
Objective:To investigate the effect of high-dose vitamin B6 on stress-induced liver cell death in rats with severe trauma and its possible mechanism.Methods:Thirty-two male SD rats were selected and divided into sham surgery group, sham surgery+B6 group, trauma group, and trauma+B6 group by using a random number table, with 8 rats in each group. Rat models of severe trauma were established by inducing abdominal wall injury, bilateral femoral fractures, unilateral cranial injury, and withdrawal of 4 ml blood from the femoral artery. The sham surgery+B6 group and trauma+B6 group were treated with saline solution plus high-dose vitamin B6, while the sham surgery group and trauma group with infusion of saline solution only. At 36 hours after injury, rat liver tissues were collected for the following experiments: (1) the genes differentially expressed in the liver tissues of the rats of the trauma group and the trauma+B6 group were screened with next-generation sequencing, followed by an analysis of the possible involvement of cell death pathways; (2) validation was conducted to ascertain whether high-dose vitamin B6 could influence various cell death pathways in the liver cells in the sham surgery group, sham surgery+B6 group, trauma group, and trauma+B6 group: apoptosis was confirmed through terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining; necroptosis was verified by mixed lineage kinase domain-like protein (MLKL) immunohistochemical staining; autophagy was examined via transmission electron microscopy; ferroptosis was confirmed by detecting oxidative malondialdehyde (MDA) levels, oxidized glutathione levels, Prussian blue staining with diaminobenzidine (DAB) enhancement, transmission electron microscopy, and immunohistochemical staining for acyl-CoA synthetase long-chain family member 4 (ACSL4); (3) Biological information analyses [Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Enrichment analysis (GSEA)] were performed for biological processes and signaling pathways represented by liver tissue sequencing results of rats between the trauma group and the trauma+B6 group.Results:(1) In the liver tissues of rats, there were 344 significantly differentially expressed genes between the trauma group and trauma+B6 group, comprising 137 upregulated genes and 207 downregulated genes, of which 18 genes were associated with apoptosis, autophagy, necroptosis, ferroptosis, and pyroptosis. (2) No significant differences were found in TUNEL staining among the sham surgery group, sham surgery+B6 group, trauma group or trauma+B6 group; MLKL protein expression levels in the liver tissues after trauma were improved, of which the trauma+B6 group was lower than that of the trauma group; Electron microscopy showed that autophagic activity in the liver cells were significantly increased after trauma, which was significantly lower of the trauma+B6 group than that of the trauma group; MDA levels in the rat liver tissues were (0.20±0.05)nmol/mg, (0.17±0.07)nmol/mg, (0.69±0.11)nmol/mg and (0.52±0.07)nmol/mg in the sham surgery group, sham surgery+B6 group, trauma group, and trauma+B6 group respectively ( P<0.01), with the trauma group having the highest MDA levels and trauma+B6 group having lower MDA levels than the trauma group; Oxidized glutathione levels in the liver tissues of the four groups were (11.75±2.09)μmol/g, (11.69±1.66)μmol/g, (19.75±3.40)μmol/g, and (14.51±1.46)μmol/g respectively ( P<0.01), with the trauma group having the highest levels and trauma+B6 group having lower levels than the trauma group; Significantly increased iron deposition was observed in the liver tissues after trauma, with lower iron deposition in trauma+B6 group than the trauma group; Electron microscopy revealed significantly lower mitochondrial membrane density in the trauma+B6 group compared to the trauma group. ACSL4 protein expression level was lower in the trauma+B6 group compared to the trauma group; (3) GO, KEGG and GSEA enrichment analyses suggested that high-dose vitamin B6 may enhance cholesterol synthesis metabolism in the liver cells and alleviate oxidative stress to reduce liver cell damage and restore normal liver cell function after trauma. Conclusions:High-dose vitamin B6 attenuates stress-induced liver injury in rats with severe trauma by inhibiting the progression of necroptosis, autophagy and ferroptosis. Its molecular mechanism may be associated with enhanced hepatic cholesterol synthesis metabolism and alleviation of oxidative stress in the liver cells.
2.Establishing prediction model of community-acquired pneumonia complicated with acute respiratory distress syndrome based on artificial neural network
Jipeng MO ; Zhongzhi JIA ; Yan TANG ; Mingxia YANG ; Hui QIN
Chinese Critical Care Medicine 2022;34(4):367-372
Objective:To investigate the independent risk factors of community-acquired pneumonia (CAP) complicated with acute respiratory distress syndrome (ARDS), and the accuracy and prevention value of ARDS prediction based on artificial neural network model in CAP patients.Methods:A case-control study was conducted. Clinical data of 414 patients with CAP who met the inclusion criteria and were admitted to the comprehensive intensive care unit and respiratory department of Changzhou Second People's Hospital Affiliated to Nanjing Medical University from February 2020 to February 2021 were analyzed. They were divided into two groups according to whether they had complicated with ARDS. The clinical data of the two groups were collected within 24 hours after admission, the influencing factors of ARDS were screened out by univariate analysis, and the artificial neural network model was constructed. Through the artificial neural network model, the importance of input layer independent variables (that was, the influence factors obtained from univariate analysis) on the output layer dependent variables (whether ARDS occurred) was drawn. The artificial neural network modeling data pairs were randomly divided into training group ( n = 290) and verification group ( n = 124) in a ratio of 7∶3. The overall prediction accuracy of the training group and the verification group was calculated respectively. At the same time, the receiver operator characteristic curve (ROC curve) was drawn, and the area under the ROC curve (AUC) was calculated. Results:All 414 patients were enrolled in the analysis, including 82 patients with ARDS and 332 patients without ARDS. Univariate analysis showed that gender, age, heart rate (HR), maximum systolic blood pressure (MSBP), maximum respiratory rate (MRR), source of admission, C-reactive protein (CRP), procalcitonin (PCT), erythrocyte sedimentation rate (ESR), neutrophil count (NEUT), eosinophil count (EOS), fibrinogen equivalent unit (FEU), activated partial thromboplastin time (APTT), total bilirubin (TBil), albumin (ALB), lactate dehydrogenase (LDH), serum creatinine (SCr), hemoglobin (Hb) and blood glucose (GLU) were significantly different between the two groups, which might be the risk factors of CAP patients complicated with ARDS. Taking the above 19 risk factors as the input layer and whether ARDS occurred as the output layer, the artificial neural network model was constructed. Among the input layer independent variables, the top five indicators with the largest influence weight on the neural network model were LDH (100.0%), PCT (74.4%), FEU (61.5%), MRR (56.9%), and APTT (51.6%), indicating that that these five indicators had a greater impact on the occurrence of ARDS in patients with CAP. The overall prediction accuracy of the artificial neural network model in the training group was 94.1% (273/290), and that of the verification group was 89.5% (111/124). The AUC predicted by the aforementioned artificial neural network model for ARDS in CAP patients was 0.977 (95% confidence interval was 0.956-1.000).Conclusion:The prediction model of ARDS in CAP patients based on artificial neural network model has good prediction ability, which can be used to calculate the accuracy of ARDS in CAP patients, and specific preventive measures can be given.
3.An analysis of the problems and countermeasures of health file construction for patients with brucellosis in Qinghai Province
Hui WAN ; Liqing XU ; Jiquan LI ; Xuxin YANG ; Li MA ; Juhua TANG ; Zhongzhi ZHAO
Chinese Journal of Endemiology 2019;38(7):578-581
The construction of health file of patients with brucellosis in Qinghai Province was based on the Qinghai Provincial Public Health Network Information Platform. It was jointly participated and implemented by animal husbandry and veterinary departments, animal disease control centers, disease control centers and hospitals at all levels, to achieve standardized management of early detection, early diagnosis, early treatment, as well as follow-up, review, and efficacy evaluation. After analyzing the health file of brucellosis patients, we proposed to strengthen the propaganda of brucellosis, collect information from multiple diseases, share resources with multiple departments, integrate human resources and exert management responsibilities, and explore opinions and countermeasures suitable for local working conditions. The establishment of a health file for patients with brucellosis could fully grasp the diagnosis and treatment of brucellosis patients, facilitate the management of follow-up and review, and it was helpful to understand the distribution and prevalence of brucellosis in Qinghai Province, and to warn the epidemic trend.
4.The study on correlation between PROC gene mutation and venous thrombosis
Zhigang ZHAO ; Wenkai ZHANG ; Bo LI ; Qing CHENG ; Guoguo ZHU ; Zhongzhi TANG
The Journal of Practical Medicine 2016;32(3):421-424
Objective To study the effect of protein C mutation on venous thrombosis (VTE) by PROC gene sequencing in patients with VTE. Methods Human PROC gene sequence was designed to amplify the third exon region of the PC-Gla domain , and then the PCR products were sequenced to search for a single nucleotide mutation (SNVs). The SNVs was constructed into eukaryotic expression system and a stable expression of wild-type and mutant PC cells were also constructed. At the same time , the distribution of PC levels in normal and VTE patients were detected with ELISA. Results Three single nucleotide mutations were found in different patients. In HUVEC cells, the synthesis of PC decreased in each mutant strain. The PC level in the normal patients , VTE patients , and the mutant samples were detected , which were significantly lower in the mutant samples than that of the VTE group (P = 0.035 3) and the normal level (P < 0.000 1). Conclusion Three mutation sites PCArg-1Cys , PCVal34Met and PCArg9Cys are important genetic factors lead to a significant decrease in plasma PC levels and the increase of VTE risk.
5.Effect of training under high temperature environment on changes in IL-17, neutro-phils and lymphocytes in peripheral blood of soldiers
Zhongzhi TANG ; Ming WU ; Xiaojuan CHEN ; Jie LIU ; Qing CHENG ; Guoguo ZHU
Military Medical Sciences 2016;40(3):199-201,206
Objective To observe the effect of a high temperature environment on IL-17 concentration , neutrophil and lymphocyte in peripheral blood of soldiers on a training task .Methods Totally 160 health training soldiers were randomly and equally divided into two groups .The test group was trained and exposed to an environment of ( 32 ±1 )℃ vs control group at ( 22 ±1 )℃ for 2 hours before the IL-17 levels in venous blood were measured and peripheral neutrophils and lymphocytes were determined.Results The body temperatures were (38.74 ±0.26),(39.23 ±0.24),and (39.51 ± 0.34)℃after training for 40 min,50 min,and 60 min under a high temperature environment , respectively, and there was a significant increase of body temperature compared to the soldiers trained under normal conditions (P <0.05).Their perceived exertion rating was 17.62 ±0.66, 18.03 ±0.56, and 18.47 ±0.84, respectively, much unlike the control group (P<0.05).After 2 hours of training under high temperature , the concentration of IL-17 was (7.12 ±4.03) pg/ml, while the number of neutrophils and lymphocytes was (6.43 ±1.27) ×109/L and (9.84 ±1.36) ×109/L, respectively in peripheral blood, significantly higher than in the control group (P <0.05).Conclusion The body temperature and rating of perceived exertion of the training soldiers are significantly elevated under a high temperature environment, and the concentration of IL-17, and the neutrophils and lymphocytes counts in peripheral blood are increased .
6.Comparison between animal temperature controller and artificial climate chamber employed for the establishment of classical heat
Huimin LI ; Xiaojuan CHEN ; Fang CHEN ; Wei FU ; Zhongzhi TANG
Chinese Journal of Emergency Medicine 2015;24(4):408-412
Objective To investigate the differences between animal temperature controller (ATC) and artificial climate chamber (ACC) used for the establishment of classical heat stroke (CHS) rat model.Methods Twenty-four male specific pathogen-free Wistar rats were randomly (random number) and equally assigned to three groups,namely room temperature control (C-C) group,heat stroke under conscious state (HS-C) group,and heat stroke under anesthesia (HS-A) group.Rats of HS-C or HS-A group were placed into ACC or ATC,then exposed to 35 ℃ heat stress.The systolic blood pressure (SBP) and core body temperature (Tc) were monitored.The time required for onset of HS was recorded.The white blood cell count (WBC) in peripheral blood and serum levels of C-reactive protein (CRP),tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) were measured.The histopathological changes of major organs were also confirmed by hematoxylin-eosin (HE) staining.Results The onset time in HS-A group was significantly shorter thanthatin HS-C group [(40.0 ± 4.3) minvs.(110.1 ± 5.3) min,P<0.01].The SBP and Tc at this moment were lower in HS-A group [(159.1 ± 5.91) mmHg vs.(174.54 ± 5.77) mmHg,P<0.01;(43.5 ± 0.4)℃ vs.(44.4 ± 0.2)℃,P<0.01].TheWBC,CRP,TNF-α and IL-1 β levels of these two HS groups were dramatically elevated compared with C-C group (P <0.01).The inflammatory cytokines levels in HS-A group were significantly lower than those in HS-C group (P < 0.01),but there was no difference in WBC between them (P > 0.05).However,there was no obvious difference in histopathological change in major organ observed between HS-A and HS-C groups.Conclusions In comparison of these two methods,ATC is similar to ACC in respect of the establishment of CHS rat model.ATC is quicker in onset of HS,and more simplified and economical than ACC and could substitute ACC.
7.The clinical value of Aescuven forte in the treatment of the mild to moderate craniocerebral trauma in the young patients
Libo LIU ; Zhongzhi TANG ; Qing CHENG ; Jing HUANG
Clinical Medicine of China 2011;27(9):970-972
ObjectiveTo assess the clinical efficacy and safety of Aescuven forte in the young patients with craniocerebral trauma.Methods Eighty patients diagnosed with craniocerebral trauma were randomized into treatment group and control group,in which the patients were given Aescuven forte tablets 0.3 g t.i.d for 30 days and routine treatmentsrespectively.Barthel index, Mini-Mental State Examination (MMSE)scale, Glasgow Coma Scale(GCS) and other clinical parameters were used to evaluate the efficacy by comparing their values before and 30 days after treatment.Results In the Aescuven forte treatment group, 35.0% (14/40) and 50.0% (20/40) of the patients showed complete response and partial response with the total response rate of 85.0% (34/40) ,while they were 20.0% (8/40) ,52.5% (21/40) and 72.5% (29/40) in the control group,respectively(x2 = 18.78 ,P < 0.05) .The incidence of complications in Aescuven forte-treated group was lower than that in the control group.No severe adverse events occurred.Conclusion Aescuven forte is a safe and effective vasoactive drug for the recovery of craniocerebral trauma-caused neurological disorders and mental deterioration in young patients.
8.Origin of some medicinal materials from animals in Chinese pharmacopoeia 2010 edition 1.
Junde LI ; Luqi HUANG ; Shihuan TANG ; Zhongzhi QIAN
China Journal of Chinese Materia Medica 2010;35(16):2052-2056
OBJECTIVEEnable to make the origin of some medicinal materials from animals-national-standard Chinese Pharmacopoeia subdivision 1 more complete, to protect and utilize the wild medicinal animal resources, and to improve the quality of traditional Chinese medicines materials materials.
METHODIn accordance with author's over two decades of studies in medicinal material from animals and medicinal animals, combining with modern animal classification findings, comparing the contents of medicinal material origin from animals in Pharmacopoeia new edition.
RESULTGive amendment advices on gallnut, concha arcae, zaocys dhumnade, gadfly, and so forth the fifteen kinds of medicinal material origin from animals' taxonomic status, Chinese names and their Latin scientific names.
CONCLUSIONThe origin of accuracy of medicinal material origin from animals is critical to ensure the quality of traditional Chinese medicine. As a national-level traditional Chinese medicine code of laws, it should advance with times, actively absorb and adopt the latest achievements in scientific research. Meanwhile, modern molecular biology techniques should be used to develop the research of conventional medicinal material origin from animals in a planned and systematic manner.
Animals ; Medicine, Chinese Traditional ; Pharmacopoeias as Topic
9.Effects of dexamethasone sodium phosphate on the proliferation of in vitro cultured rabbit articular chondrocytes
Guoxin LI ; Chenfeng TANG ; Jian WEN ; Zhongzhi YUAN
Chinese Journal of Tissue Engineering Research 2009;13(50):9984-9987
BACKGROUND: Studies have demonstrated that dexamethasone can reduce type Ⅱ collagen and increase type I collagen in articular cartilage surface matrix, but the action mechanism of glucocorticoid to articular chondrocyte proliferation remains unclear. OBJECTIVE: To investigate the effects of dexamethasone sodium phosphate on rabbit articular chondrocytes cultured in vitro. DESIGN: Comparative observation.MATERIALS: New Zealand rabbits, 1 month old, were used for chondrocyte isolation and culture.METHODS: Rabbit articular chondrocytes cultured in vitro were randomly divided into control and experimental groups. Cells of the control group were cultured in DMEM media without dexamethasone sodium phosphate. Cells of experimental groups were cultured in DMEM media with different concentrations of dexamethasone sodium phosphate (0.02, 0.1, 0.5 g/L), respectively. MAIN OUTCOME MEASURES: Primary and passage chondrocytes were observed. Flow cytometry and immunocytochemistry were used to observe the effect of dexamethasone sodium phosphate on cell proliferation and type Ⅱ collagen synthesis. The ultrastructural changes of cultured chondrocytes were observed by transmission electron microscopy. RESULTS: The attachment and proliferation of experimental group chondrocytes was slower than control group. There average gray scale values of the experimental groups were significantly greater than the control group (P < 0.05). The cellular proportions of S phase and G_2+M phase of the experimental groups decreased but the cellular proportions of G0/G1 period increased. Under transmission electron microscope the amount of rough endoplasmic reticulum and mitochondria of the experimental groups decreased. CONCLUSION: Dexamethasone sodium phosphate inhibited articular chondrocyte proliferation, possibly due to the decrease of type Ⅱcollagen and protein synthesis ability of chondrocyte.
10.Bone grafting and internal fixation for treating fibrous dysplasia of the adult proximal femur A 13.case report
Zhongzhi YUAN ; Chenfeng TANG ; Guoxin LI ; Jian WEN ; Gang LIU ; Yunzhuo YANG ; Jiyun LI
Chinese Journal of Tissue Engineering Research 2009;13(30):5985-5990
The present study retrospectively analyzed 13 patients with fibrous dysplasia of the adult proximal femur who received treatment at the Department of Orthopedics,Shenzhen Hospital,Peking University between March 2000 and June 2006.These patients comprised 6 males and 7 females,with an age of 21-35 years.Among 11 patients suffering from mild hip pain,6 presented with mild limping due to worsened pains,and 2 exhibited obvious manifestations of bone fracture.All patients underwent X-ray,CT examinations,and 3 were subjected to MRI examinations,prior to bone grafting and internal fixation.Monostotic lesion was observed in 10 patients and polystotic lesion in 3 patients,but endocrine disturbance was not observed in any patient.Nine patients suffered from lesions in the femoral neck and 4 from lesions in the femoral rotator.Curettage,autogenous and allogeneic bone grafting,and internal fixation with dynamic hip screws and proximal femoral nails were successfully performed in each patient.Internal fixation time averaged 165 minutes (range 120-210 minutes).During the early stage after surgery,weight-bearing activities were not permitted.Bone fracture healed after an average of 3 months.Eighteen to forty-eight months of follow-up demonstrated that clinical symptoms were relieved,with good hip joint function and basically normal gait.X-ray plains showed thickened cortical bone,as well as compacted ossification shadow in bone grafting region.In addition,partial bone absorption,but not recurrent clinically,was observed in 4 patients.These findings imply that thorough curettage,autogenous and allogeneic bone grafting,and internal fixation with dynamic hip screws and proximal femoral nails is a reliable method to treat fibrous dysplasia of the adult proximal femur and it can acquire satisfactory curative effects.

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