1.Analysis of Tongue Coating Microbiota Characteristics in Coronary Heart Disease with Qi Deficiency and Blood Stasis Syndrome
Chuhao WANG ; Yongyue LIU ; Zhaoxuan DING ; Xiaoqing ZHANG
Journal of Traditional Chinese Medicine 2025;66(5):501-508
ObjectiveTo explore the characteristics of the tongue coating microbiota in patients of coronary heart disease (CHD) with qi deficiency and blood stasis syndrome. MethodsA total of 27 CHD patients with qi deficiency and blood stasis syndrome, 29 patients with non-qi deficiency and blood stasis syndrome, and 20 healthy individuals were included in this study. The tongue coating microbiota of the participants was analyzed using 16S rDNA high-throughput sequencing technology, followed by Alpha and Beta diversity analyses and comparisons of microbial abundance. ResultsA total of 479 operational taxonomic units (OTUs) were detected, among which 245 OTUs were shared across all three groups. There were 33 OTUs unique to the qi deficiency and blood stasis syndrome group, 21 OTUs unique to the non-qi deficiency and blood stasis syndrome group, and 121 OTUs unique to the healthy group. The observed species count (Sob), total species richness (Chao1), abundance-based coverage estimator (ACE), and Shannon diversity index were significantly lower in the qi deficiency and blood stasis syndrome and non-qi deficiency and blood stasis syndrome groups compared to the healthy group (P<0.05). Principal coordinate analysis (PCoA) of the tongue coating microbiota showed significant differences in distance matrices among the three groups (P<0.05). Compared with the healthy group, the qi deficiency and blood stasis syndrome group exhibited an increased abundance of Actinobacteria, Patescibacteria, Spirochaetes, Verrucomicrobia, Rothia, TM7X, Gemella, and Corynebacterium, while Fusobacteria, Cyanobacteria, Leptotrichia, and Lactobacillus decreased (P<0.05). In the non-qi deficiency and blood stasis syndrome group, Actinobacteria, Verrucomicrobia, Rothia, and Corynebacterium increased, whereas Cyanobacteria and Lactobacillus reduced (P<0.05). When comparing with the non-qi deficiency and blood stasis syndrome group, the qi deficiency and blood stasis syndrome group had a significantly higher abundance of Patescibacteria, Peptostreptococcus, Solobacterium, Filifactor, Moraxella, Porphyromonas endodontalis, and Capnocytophaga, while Cyanobacteria reduced (P<0.05). Conclusuion Patients with CHD of qi deficiency and blood stasis syndrome exhibit a decrease in beneficial bacteria and an increase in pathogenic bacteria. Patescibacteria, Peptostreptococcus, Solobacterium, Filifactor, Moraxella, Porphyromonas endodontalis, and Capnocytophaga were identified as the key differential microbiota distinguishing qi deficiency and blood stasis syndrome from non-qi deficiency and blood stasis syndrome patients.
2.The assessment of hepatic hemodynamic characteristics in cirrhotic patients with splenectomy based on iodine map of dual-source CT
Yongyue ZHU ; Zhou ZHOU ; Chengli ZHAO ; Yinghan SUN ; Cong ZHOU ; Daoqing WANG
Journal of Practical Radiology 2024;40(2):302-305
Objective To investigate the hepatic hemodynamic characteristics of cirrhotic patients with splenectomy using iodine map of dual-source computed tomography(DSCT).Methods Twenty-four cirrhotic patients with splenectomy were selected as a study group,41 cirrhotic patients without splenectomy as a cirrhosis group and other 32 patients with normal liver as a control group.The iodine concentration(IC)in hepatic arterial and venous phases was measured on the iodine map,and the arterial iodine fraction(AIF)and portal venous iodine concentration(PVIC)were calculated.Receiver operating characteristic(ROC)curves were plotted and the area under the curve(AUC)was recorded to evaluate the diagnostic efficacy of each parameter using the DeLong test.Results IC in arterial phase and AIF were significantly higher,and IC in venous phase and PVIC were significantly lower in study group(P<0.05).The AUC values of the four parameters between study group and cirrhosis group were 0.735,0.992,0.943,and 0.994,respectively.Conclusion DSCT iodine map is helpful for clinical quantitative assessment of hepatic hemodynamic characteristics in cirrhotic patients with splenectomy,and the PVIC has optimal independent diagnostic performance.
3.Epidemiological survey of 2019-nCoV infection in staff and students in some public health schools in China
Yongyue WEI ; Wenjing GAO ; Longyao ZHANG ; Shaoguan WANG ; Siyan ZHAN ; Tao REN ; Yuantao HAO ; Jun LYU ; Liming LI
Chinese Journal of Epidemiology 2023;44(2):175-183
Objective:To understand the infection status and characteristics of 2019-nCoV infection in different areas in China after the adjustment of the national prevention and control strategy of 2019-nCoV infection.Methods:The online questionnaire survey was conducted among staff and students of 39 public health schools in 23 provinces (municipalities) in China from 12: 00 on December 20 to 9: 00 on December 23, 2022. The infection rates in staff and students in all the provinces were estimated. The risk factors, demographic and clinical characteristics of 2019-nCoV infections were explored.Results:A total of 28 901 valid questionnaires were obtained (26 355 from students and 2 546 from staff) with a qualified rate of 94.3%. The infection rates varied greatly among provinces and cities; the infection rates in students and staff in Beijing reached 78.55% and 76.40%, respectively. Infection rates in students and staff in Tianjin and Hebei also exceeded 65.00%, and 96.76% of infections occurred on and after December 1, 2022. Students had lower risk for the infection compared with staff ( OR=0.72, 95% CI: 0.60-0.86). Compared with age group ≤20 years, the OR of age groups 21-30, 31-40, 41-50, 51-60 and > 60 years were 1.22 (95% CI: 1.14-1.30), 1.54 (95% CI: 1.30-1.84), 1.25 (95% CI: 0.99-1.58), 1.29 (95% CI: 0.94-1.78) and 1.19 (95% CI: 0.51-2.80), respectively. The longer the period after the last vaccination, the higher the risk for the infection. Compared with those who received the last vaccination in the past 3 months, the OR of those who received the last vaccination in the past 4-6 months, 7-9 months, 10-12 months, 13-15 months and ≥16 months were 1.56 (95% CI: 1.34-1.82), 1.59 (95% CI: 1.36-1.86), 1.67 (95% CI: 1.45-1.93), 1.86 (95% CI:1.58-2.19) and 2.46 (95% CI: 2.09-2.90), respectively. Compared with those living alone, the OR of those living with 1-2, 3-4 and ≥5 roommates were 17.55 (95% CI: 15.91-19.39), 20.22 (95% CI: 18.25-22.43) and 11.78 (95% CI: 10.40-13.36), respectively. Only 5.94% of the staff and 7.19% of the students reported asymptomatic infections. Among those with symptoms, 88.18% of students and 85.65% of staff reported symptom of fever. Conclusions:The transmission dynamics of 2019-nCoV infection varied significantly across the country. The speed of transmission of 2019-nCoV and clinical severity of the infection were far beyond our knowledge. Organized epidemiological survey should be regularly carried out to provide reliable data support for more accurate prediction of the epidemic and medical resource allocation.
4.Advances in several important antimicrobial lipopeptids from Bacillus spp.
Daoming LI ; Ying WANG ; Chao CHEN ; Mingbai ZENG ; Qianru LI ; Qingyun JIA ; Xiuli LIU ; Yongyue HOU ; Chengming FAN ; Yuhong CHEN ; Zanmin HU
Chinese Journal of Biotechnology 2022;38(5):1768-1783
Bacillus spp. are probiotics and can secrete a variety of natural antimicrobiol active substances, of which lipopeptides are an important class. Up to now, about 90 lipopeptides have been identified, and most of them are cyclic lipopeptides. surfactin, iturin, fengycin, bacillomycin and polymyxins are widely studied, and the first three have huge potential for application due to their properties of surfactants and anti-fungal, anti-bacterial, anti-viral, anti-tumor and anti-inflammatory functions. In this paper, the research progress in the structure, function, synthesis regulation, separation, purification and production of surfactin, iturin and fengycin was reviewed. Synthetic biology is a vital means to increase the yield of lipopeptides, and in the future, lipopeptides can be used in crop cultivation, animal farming, food, medicine and petroleum industries as well as environmental protection. Future research should be strengthened on the discovery of new lipopeptides, synthesis of high-activity lipopeptides, economical production of lipopeptides on a large scale and their safety evaluation.
Anti-Bacterial Agents
;
Anti-Infective Agents/pharmacology*
;
Bacillus
;
Bacillus subtilis
;
Lipopeptides/pharmacology*
;
Peptides, Cyclic/pharmacology*
5.Inhibitory effect of RMT1-10-induced tolerogenic dendritic cells in vitro on high-risk corneal allograft rejection in mice and its mechanism
Min ZHAO ; Liuqing YANG ; Mengyu WANG ; Yu TAO ; Yongyue GUO ; Ruifeng SU ; Jing SHI ; Xiaobo TAN
Chinese Journal of Experimental Ophthalmology 2022;40(8):725-733
Objective:To investigate the inhibitory effect of RMT1-10-induced tolerogenic dendritic cells (Tol-DCs) in vitro on high-risk corneal allograft rejection in mice and its mechanism. Methods:One hundred SPF male BALB/c mice and fifty SPF male C57BL/6 mice were selected.Bone marrow-derived immature dendritic cells (imDCs) obtained from C57BL/6 mice were divided into imDCs group, mature dentritic cells (mDCs) group, RMT1-10 group, and IgG isotype control group.The imDCs in the four groups were cultured with no intervention, lipopolysaccharide, RMT1-10 and lipopolysaccharide, or IgG isotype antibody and lipopolysaccharide for 7 days according to grouping.The expression levels of different phenotypes of DCs including CD11c, CD80, CD86, major histocompatibility complex (MHC)-Ⅱ, T cell immunoglobulin and mucin domain containing molecule (Tim)-4 and CD103 in the four groups were detected by flow cytometry.The concentrations of interleukin-10 (IL-10) and transforming growth factor-β (TGF-β) in the DCs supernatants were determined by enzyme-linked immunosorbent assay.A mixed lymphocyte culture system was established, and the stimulation index (SI) of CD4 + T cell proliferation stimulated with DCs was detected by cell counting kit 8 method.Corneal neovascularization was induced by corneal stromal suture in BALB/c mice, and the 80 mice with neovascularization in 4 quadrants growing into the middle and peripheral cornea were used as recipients.The recipient mice were randomized into imDCs group, mDCs group, RMT1-10 group, and IgG isotype control group using the random number table method, with 20 mice in each group.An injection of corresponding DCs (1×10 6 cells/100 μl) was administered to the recipient mice via the tail vein according to grouping.At 7 days following the injection, C57BL/6 mice were used as donors and penetrating keratoplasty was performed.Within one month after the operation, signs of corneal grafts rejection, including opacity, edema and neovascularization, were observed by slit lamp biomicroscopy and scored every day.At 21 days after the operation, 5 recipients selected from each group were subcutaneously injected with naive C57BL/6 splenocytes (1×10 6 cells/100 μl) behind the ear.The delayed type hypersensitivity (DTH) was evaluated by ear swelling at 24 hours after the subcutaneous injection.The use and care of experimental animals complied with the Regulations on the Management of Experimental Animals promulgated by the State Science and Technology Commission.This study protocol was approved by an Ethics Committee of the Affiliated Hospital of Chengde Medical University (No.CYFYLL2020055). Results:Compared with mDCs group, the expressions of CD80, CD86 and MHC-Ⅱ, and the percentage of Tim-4-positive cells in CD11c-positive cells were significantly decreased in RMT1-10 group, showing statistically significant differences (all at P<0.001). The percentage of Tim-4-positive cells were significantly decreased in RMT1-10 group than imDCs group, and the percentage of CD103-positive cells in RMT1-10 group was significantly higher than imDCs group, mDCs group and IgG isotype control group (all at P<0.001). The concentrations of IL-10 and TGF-β in the cell culture supernatant of RMT1-10 group were significantly higher than those of the other three groups, with statistically significant differences (all at P<0.001). There were statistically significant differences in the SI of CD4 + T cell proliferation simulated by DCs ( Fgroup=1 833.00, P<0.001; Fratio=230.40, P<0.001; Finteraction=3.06, P=0.01). The SI of DCs/CD4 + T cells ratio at 1∶5, 1∶10, 1∶20 and 1∶40 were all significantly lower in imDCs group than mDCs group, and were all significantly lower in RMT1-10 group than imDCs group (all at P<0.05). There was a statistically significant difference in corneal graft survival curve among various groups ( χ2=77.69, P<0.001). The survival rate of RMT1-10 group was significantly higher than that of imDCs group ( χ2=9.74, P=0.002), and the survival rate of imDCs group was significantly higher than that of mDCs group ( χ2=31.02, P<0.001). The ear swelling of recipient mice of positive control group, mDCs group, IgG isotype control group, imDCs group and RMT1-10 group was (503.6±17.2), (475.7±17.6), (456.2±18.8), (225.2±39.4), (118.1±12.6), and (106.4±7.4) μm, with a statistically significant difference among them ( F=377.10, P<0.001). The mice ear swelling was more serious in positive control group than mDCs group, more serious in IgG isotype control group than imDCs group, and more serious in imDCs group than RMT1-10 group (all at P<0.05). Conclusions:RMT1-10 can inhibit the rejection of high-risk corneal transplantation in mice, the mechanism of which may be attributed to inducing imDCs to transform into Tol-DCs in vitro and up-regulating the expression of TGF-β and IL-10, which promotes antigen-specific immune tolerance after adoptive transfer, thereby indirectly prolongs the survival of corneal grafts.
6.Effects of parathyroidectomy on heart rate circadian rhythm in patients with stage 5 chronic kidney disease combined with severe secondary hyperparathyroidism
Ying CUI ; Hui HUANG ; Wenkai REN ; Guang YANG ; Ming ZENG ; Xiaoming ZHA ; Shaowen TANG ; Yaoyu HUANG ; Zhanhui GAO ; Fangyan XU ; Wenbin ZHOU ; Hanyang QIAN ; Jing WANG ; Chun OUYANG ; Xueyan GAO ; Changying XING ; Yongyue WEI ; Ningning WANG
Chinese Journal of Nephrology 2021;37(5):414-423
Objective:To observe heart rate circadian rhythm in patients with chronic kidney disease (CKD) stage 5 and to analyze the effects of parathyroidectomy (PTX) on heart rate circadian rhythm in severe secondary hyperparathyroidism (SHPT) patients.Methods:A cross-sectional observation was performed in 213 patients with CKD stage 5 and 96 controls, and the patients were divided into those with severe SHPT (PTX group, n=70) and without severe SHPT (non-PTX group, n=143). Forty-six PTX patients were followed up prospectively. The baseline data were compared among these groups. Holter electrocardiogram was performed for each participant. Non-dipping heart rate was defined as night/day heart rate ratio greater than 0.9. Multiple linear regression analysis was used to analyze the related factors of heart rate circadian rhythm in patients with CKD stage 5. Results:The 24-hour, daytime and nighttime mean heart rate in patients with CKD stage 5 were all higher than those in controls, especially in PTX group (all P<0.05). The night/day heart rate ratios of controls and CKD stage 5 patients were (0.81±0.08) and (0.91±0.08) respectively ( P<0.01). Correlation analysis showed 24-hour and daytime or nighttime mean heart rate in patients with CKD stage 5 were positively correlated with serum levels of phosphorus and ln(alkaline phosphatase), while nighttime mean heart rate and night/day heart rate ratio were positively related with serum intact parathyroid hormone level. After adjusting with postoperative follow-up period (median time: 10.9 months), 24-hour and nighttime mean heart rate, and night/day heart rate ratio in PTX patients all decreased significantly (all P<0.01). Conclusions:Heart rate is increased and circadian rhythm is abnormal in patients with CKD stage 5, which are related with mineral and bone disorder. PTX significantly decreases 24-hour and nighttime mean heart rate in severe SHPT patients, and improves the heart rate circadian rhythm.
7.Value of abnormal circadian rhythm of heart rate predicting the all-cause mortality in stage 5 chronic kidney disease patients
Wenkai REN ; Ying CUI ; Ming ZENG ; Hui HUANG ; Shaowen TANG ; Guang YANG ; Yaoyu HUANG ; Zhanhui GAO ; Fangyan XU ; Hanyang QIAN ; Jing WANG ; Chun OUYANG ; Xueyan GAO ; Yifei GE ; Yujie XIAO ; Changying XING ; Yongyue WEI ; Ningning WANG
Chinese Journal of Nephrology 2021;37(7):558-566
Objective:To investigate the predictive value of abnormal heart rate circadian rhythm for all-cause mortality in stage 5 chronic kidney disease (CKD 5) patients.Methods:The retrospective study was performed in CKD 5 patients enrolled from the First Affiliated Hospital of Nanjing Medical University (Jiangsu Province Hospital) and the Affiliated BenQ Hospital of Nanjing Medical University from February, 2011 to December, 2019. A total of 159 healthy volunteers were enrolled as the healthy control group during the same period. The circadian rhythm of heart rate was monitored by 24-hour Holter. Related indices (including 24-hour, daytime and nighttime mean heart rate, night/day heart rate ratio, 24-hour maximum heart rate, 24-hour minimum heart rate and difference between maximum and minimum of 24-hour heart rate) were calculated. Non-dipping heart rate was defined as night/day heart rate ratio greater than 0.9. Cox regression model was used to analyze the risk factors of all-cause mortality in CKD 5 patients. Kaplan-Meier survival curve and Log-rank test were used to compare the differences of cumulative mortality between high ratio group (night/day heart rate ratio>0.91) and low ratio group (night/day heart rate ratio≤0.91). The nonlinear relationship between night/day heart rate ratio and all-cause mortality was analyzed by restricted cubic spline plot. Time-dependent receiver operating characteristic (ROC) curve was used to analyze the predictive value of night/day heart rate ratio for all-cause mortality in CKD 5 patients.Results:A total of 159 healthy volunteers and 221 CKD 5 patients were included in this study. There were 123 males (55.66%) and the age was (52.72±13.13) years old in CKD 5 patients. The total median follow-up time was 50.0 months. Compared with controls, 24-hour, nighttime mean heart rate, 24-hour minimum heart rate in CKD 5 patients were increased (all P<0.05), furthermore, the night/day heart rate ratio was higher [(0.91±0.09) vs (0.81±0.08), P<0.001], showing "non-dipping heart rate". However, the 24-hour maximum heart rate and the difference between maximum and minimum of 24-hour heart rate in CKD 5 patients were lower than controls (both P<0.05). Multivariate Cox regression analysis showed that the increased night/day heart rate ratio (per 0.1 increase, HR=1.557, 95% CI 1.073-2.258, P=0.020) was an independent influencing factor for all-cause mortality in CKD 5 patients. Kaplan-Meier survival curve analysis showed that the cumulative mortality of the high ratio group was significantly increased than that of the low ratio group (Log-rank test χ 2=7.232, P=0.007). From the restricted cubic spline plot, there was a linear effect between night/day heart rate ratio and all-cause mortality ( P=0.141), and when night/day heart rate ratio was above 0.91, the risk of all-cause mortality was significantly increased in CKD 5 patients. According to time-dependent ROC curve, the accuracy of night/day heart rate ratio in predicting all-cause mortality was 70.90% even when the survival time was up to 70.0 months. Conclusions:The circadian rhythm of heart rate in CKD 5 patients displays "non-dipping" state. High night/day heart rate ratio is an independent influencing factor for all-cause mortality in CKD 5 patients.
8. Fitting and forecasting the trend of COVID-19 by SEIR+ CAQ dynamic model
Yongyue WEI ; Zhenzhen LU ; Zhicheng DU ; Zhijie ZHANG ; Yang ZHAO ; Sipeng SHEN ; Bo WANG ; Yuantao HAO ; Feng CHEN
Chinese Journal of Epidemiology 2020;41(4):470-475
Objectives:
Fitting and forecasting the trend of COVID-19 epidemics.
Methods:
Based on SEIR dynamic model, considering the COVID-19 transmission mechanism, infection spectrum and prevention and control procedures, we developed SEIR+ CAQ dynamic model to fit the frequencies of laboratory confirmed cases obtained from the government official websites. The data from January 20, 2020 to February 7, 2020 were used to fit the model, while the left data between February 8-12 were used to evaluate the quality of forecasting.
Results:
According to the cumulative number of confirmed cases between January 29 to February 7, the fitting bias of SEIR+ CAQ model for overall China (except for cases of Hubei province), Hubei province (except for cases of Wuhan city) and Wuhan city was less than 5%. For the data of subsequent 5 days between February 8 to 12, which were not included in the model fitting, the prediction biases were less than 10%. Regardless of the cases diagnosed by clinical examines, the numbers of daily emerging cases of China (Hubei province not included), Hubei Province (Wuhan city not included) and Wuhan city reached the peak in the early February. Under the current strength of prevention and control, the total number of laboratory- confirmed cases in overall China will reach 80 417 till February 29, 2020, respectively.
Conclusions
The proposed SEIR+ CAQ dynamic model fits and forecasts the trend of novel coronavirus pneumonia well and provides evidence for decision making.
9.Dexmedetomidine-induced cardioprotection in a mouse model of lung ischemia-reperfusion: the relationship with endoplasmic reticulum stress
Bingqian XIANG ; Ziyin LUO ; Hui GAO ; Yongyue DAI ; Wantie WANG
Chinese Journal of Anesthesiology 2017;37(1):61-65
Objective To evaluate dexmedetomidine-induced cardioprotection in a mouse model of lung ischemia-reperfusion (I/R) and the relationship with endoplasmic reticulum stress.Methods Forty healthy SPF male C57BL/6J mice,weighing 20-24 g,aged 8-10 weeks,were divided into 4 groups (n=10 each) using a random number table:sham operation group (Sham group),lung I/R group (I/R group),dexmedetomidine group (Dex group) and dexmedetomidine plus atipamezole (specific α2-adrenergic receptor antagonist) group (DA group).The model of lung I/R injury was established by clamping the left hilum of lung for 30 min followed by 180 min of reperfusion.In group Sham,only sternotomy was performed,the hilum of lung was not clamped,and the mice were mechanically ventilated for 210 min.In Dex and DA groups,dexmedetomidine 20 μg/kg and dexmedetomidine 20 μg/kg plus atipamezole 250 μg/kg were injected intraperitoneally,respectively,at 30 min before establishment of the model.At 180 min of reperfusion,blood samples were collected from the orbit for determination of creatine kinase-MB (CK-MB) and lactic dehydrogenase (LDH) activities in serum.The animals were then sacrificed,and hearts were removed for determination of apoptosis in cardiomyocytes (by TUNEL) and expression of phosphorylated c-Jun N-terminal kinase (p-JNK),caspase-12,CCAAT/enhancer-binding protein homologous protein (CHOP) and glucose-regulated protein 78 (GRP78) in myocardial tissues (by Western blot),and expression of JNK,caspase-12,CHOP,GRP78 mRNA in myocardial tissues (by real-time polymerase chain reaction).Apoptosis index was calculated.Results Compared with Sham group,the serum CKMB and LDH activities and apoptosis index were significantly increased,the expression of p-JNK,JNK mRNA,and caspase-12,CHOP and GRP78 protein and mRNA was up-regulated in I/R,Dex and DA groups (P<0.01).Compared with I/R group,the serum CK-MB and LDH activities and apoptosis index were significantly decreased,the expression of p-JNK,JNK mRNA,and caspase-12 and CHOP protein and mRNA was down-regulated,the expression of GRP78 protein and mRNA was up-regulated in group Dex,and the expression of GRP78 protein and mRNA was significantly up-regulated (P<0.01),and no significant change was found in the other parameters in group DA (P>0.05).Compared with DEX group,the serum CK-MB and LDH activities and apoptosis index were significantly increased,the expression of pJNK,JNK mRNA,and caspase-12 and CHOP protein and mRNA was up-regulated (P<0.01),and no significant change was found in the expression of GRP78 protein and mRNA in DA group (P>0.05).Conclusion Dexmedetomidine can reduce myocardial injury induced by lung I/R,and the mechanism may be related to activation of α2-adrenergic receptors and inhibition of endoplasmic reticulum stress in myocardial cells of mice.
10.Effects of traditional Chinese medicine Yiqi Huoxue Tongluo Jiedu prescription on pneumocyte apoptosis and c-Jun N-terminal protein kinase pathway in mice after lung ischemia/reperfusion injury
Lu SHI ; Dong SONG ; Xuguang JIA ; Ziyin LUO ; Bingqian XIANG ; Yongyue DAI ; Min LUO ; Wantie WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2017;24(1):73-77
Objective To observe the effects of Yiqi Huoxue Tongluo Jiedu fang (YHTJF) on pneumocyte apoptosis after lung ischemia/reperfusion (I/R) injury (LIRI) in mice and to investigate whether c-Jun N-terminal protein kinase (JNK) is involved in the mechanism of apoptosis.Methods Seventy C57BL/6J male mice were randomly divided into seven groups:normal control group (C group),carboxyl methyl cellulose-Na+normal control group (CMC-Na+C group),CMC-Na+sham group (CMC-Na+S group),CMC-Na+I/R group (CMC-Na+I/R group) and CMC-Na+YHTJF-low,-middle,-high dose groups (CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups).C group did not undergo any processing;in CMC-Na+S group,only was chest opened without clipping the lung hilum;in the rest of the four groups,they all underwent opening of the chest and clipping the lung hilum for 30 minutes,then the clipping of artery was relieved and left lung reperfusion was carried out for 3 hours.After operation,the mice were sacrificed,the lung tissues were harvested.Under light and electron microscopes,the lung morphological and ultra-structural changes were observed,and the changes of index of quantitative evaluation for alveolar damage (IQA) were determined.The terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) was applied to evaluate the apoptosis index (AI) of the lung tissues.The protein and mRNA expressions of JNK and glucose regulating protein 78 (GRP78) in lung tissues were detected by Western Blot and reverse transcription-polymerase chain reaction (RT-PCR);the correlations between lung AI and the expressions of mRNA and protein of JNK and GRP78,IQA were analyzed.Results Compared with CMC-Na+S group,IQA,AI and mRNA and the protein expressions of JNK and GRP78 in CMC-Na+I/R group were obviously higher [IQA:(74.00 ± 7.31)% vs.(7.00 ± 1.23)%,AI:(64.40 ± 11.97)% vs.(5.60 ± 1.14)%,JNK mRNA (gray value):1.143 ± 0.284 vs.0.152 ± 0.128,GRP78 mRNA (gray value):0.897 ± 0.129 vs.0.284 ± 0.044,JNK protein (A value):0.428 ± 0.074 vs.0.073 ± 0.052,GRP78 protein (A value):1.075 ± 0.145 vs.0.589 ± 0.060].Compared with CMC-Na+I/R group,the IQA,AI,protein and mRNA expressions of JNK and GRP78 in CMC-Na+YL,CMC-Na+YM,CMC-Na+YH groups were all lower,and the degree of reduction in group CMC-Na+YM was the most remarkable,greater than that in CMC-Na+YL or CMC-Na+YH group [IQA:(26.20 ± 3.35)% vs.(34.00±5.34)%,(41.20±9.18)%,AI:(29.40±3.05)% vs.(48.20±3.83)%,(39.20±6.14)%,JNK mRNA (gray value):0.681 ± 0.130 vs.0.804 ± 0.153,0.938 ± 0.11,GRP78 mRNA (gray value):0.450 ± 0.105 vs.0.747 ± 0.231,0.566 ± 0.115,JNK protein (A value):0.188 ± 0.049 vs.0.261 ± 0.065,0.209 ± 0.063,all P < 0.01],compared with the CMC-Na+I/R group,the expression of GRP78 protein was obviously higher in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkably high was in CMC-Na+YH group (A value:1.429 ±0.226 vs.1.130±0.169,1.128 ±0.177,all P < 0.01).The apoptosis of each group was mainly in the pulmonary vascular endothelial cells and alveolar epithelial cells,and brown particles were positive cells under light microscope.Under transmission electron microscope:nuclear pyknosis and margination under the nuclear membrane,cytoplasm condensed,lamellar bodies decreased and emptying increased,cell membrane microvilli decreased or disappeared,mitochondria swelling,inflammatory cells increased in alveolar septum and adhering onto the capillary walls could be seen in CMC-Na+I/R group.Compared with CMC-Na+I/R group,the lung tissue ultrastructural damage alleviated,ultrastructure of alveoli clearly seen,nuclear chromatin relatively uniform,cytoplasm increased,type Ⅱ alveolar epithelial cell surface microvilli relatively plenty,lamellar corpuscle number increased,mitochondria swelling ameliorated in CMC-Na+YH,CMC-Na+YL,CMC-Na+YM groups and the most remarkable one was CMC-Na+YM group.AI was significantly positive correlated with the mRNA and protein expressions of JNK,GRP78 and IQA (r =0.907,0.928,0.880,0.712,0.911,all P < 0.01).Conclusions YHTJF may effectively alleviate the cell apoptosis in mice LIRI,and its mechanism may be related to the inhibition of JNK pathway.

Result Analysis
Print
Save
E-mail