1.Effects of Serum Containing Wenjing Tongluo Decoction on Mouse Chondrocyte Injury and IκB-ζ/HIF-1α/LDHA Axis
Wei WEI ; Chenjian PENG ; Renjun GU ; Xiwu YAN ; Jiapeng YE ; Guicheng HUANG ; Luning SUN
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(5):469-478
OBJECTIVE To study the effects and mechanisms of serum containing Wenjing Tongluo Decoction on interleukin-1β(IL-1β)-induced mouse primary chondrocyte injury.METHODS The IL-1β-induced primary chondrocyte injury model was estab-lished,and the mRNA expression of inflammatory factors IL-1β,IL-6,TNF-α,cartilage degradation-related proteases matrix metal-loproteinase 3(MMP3),MMP9,MMP13,ADAM metallopeptidase containing thrombospondin 1 motif 4(ADAMTS4),and glycoly-sis-related enzymes lactate dehydrogenase A(LDHA),M2 pyruvate kinase(PKM2),reduced coenzyme Ⅱ oxidase 2(NOX2)and re-duced coenzyme Ⅱ oxidase 4(NOX4)in the cells was detected;the intracellular MMP3,MMP13,P65,IκB-ζ,hypoxia-inducible factor 1α(HIF-1α)and LDHA protein expression levels were determined;the concentrations of nitric oxide(NO),malondialdehyde(MDA)and lactic acid in the cell supernatant were further detected,and the ratio of the reduced state(NADH)and oxidized state(NAD+)of intracellular coenzyme Ⅰ(NAD),as well as intracellular reactive oxygen species(ROS)levels were assayed.RESULTS Serum containing Wenjing Tongluo Decoction significantly inhibited the mRNA expression of IL-1β,IL-6,and TNF-α in IL-1β chondrocytes(P<0.01),reduced the NO concentration in the cell supernatant(P<0.05,P<0.01),and downregulated the protein ex-pression of IκB-ζ(P<0.05)and P65(P<0.05,P<0.01).Serum containing Wenjing Tongluo Decoction significantly downregulated the mRNA expression of MMP3,MMP9,MMP13 and ADAMTS4(P<0.01),and inhibited the protein expression of MMP13(P<0.05,P<0.01)and MMP3(P<0.05).In terms of oxidative stress,it significantly suppressed the production of ROS in chondrocytes,increased the NADH/NAD+ratio,reduced the MDA concentration in the supernatant,and downregulated the expression of HIF-1α protein(P<0.01).The serum containing Wenjing Tongluo Decoction significantly reduced the lactic acid concentration,downregulated the expression of LDHA,PKM2,NOX2 and NOX4,and reduced the level of intracellular glycolysis(P<0.05,P<0.01).CONCLU-SION The serum containing Wenjing Tongluo Decoction can alleviate IL-1β-induced mouse primary chondrocyte injury by inhibiting inflammation,cartilage degradation,oxidative stress and glycolysis,which may be related to the regulation of IκB-ζ/HIF-1α/LDHA axis.
2.Study on the Mechanism of Banxia Xiexin Decoction in Treating Functional Dyspepsia
Defang ZOU ; Renjun GU ; Mingxi ZHU ; Lang REN ; Ruizhi TAO ; Keqin LU ; Aiyun WANG ; Zhiguang SUN
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(9):941-948
OBJECTIVE To evaluate the effects of different functional dyspepsia(FD)modeling methods and explore the thera-peutic effect and potential mechanism of Banxia Xiexin Decoction on FD.METHODS BALB/c mice were randomly divided into the blank group,iodoacetamide group,loperamide group,tail clamp group and vinegar group.After 1 week of intervention,the status of mice in each group was observed and their gastrointestinal motility,hormone levels and pathological changes were detected.A more i-deal FD modeling method was evaluated and determined.After modeling,different doses of Banxia Xiexin Decoction were given to in-tervene,and the changes in the gastrointestinal function of mice were observed.The expression of related proteins was studied by im-munohistochemistry,ELISA,Western Blot and other experimental methods.RESULTS Comparing the four modeling methods,it was found that the mice in the iodoacetamide group,loperamide group,and vinegar group showed weight loss compared to the blank group;the gastric emptying rate and small intestinal propulsion rate of mice in the iodoacetamide group and vinegar group decreased;changes in gastrointestinal hormones were found in the serum of mice in the tail clip group and vinegar group.Finally,the iodoacetamide meth-od was evaluated as the optimal FD modeling method.The administration results showed that Banxia Xiexin Decoction had no signifi-cant effect on the food intake and body weight of FD mice,while medium and high doses could improve the physical condition of FD mice,increase their gastric emptying rate and small intestine propulsion rate.The experimental results of immunohistochemistry,West-ern blotting,and ELISA confirmed that medium and high doses of Banxia Xiexin Decoction can significantly reduce the expression lev-els of TNF-α and IL-6 in the duodenum and serum of FD mice.CONCLUSION The iodoacetamide method is a better FD modeling method.Banxia Xiexin Decoction can improve the condition of FD mice,increase gastrointestinal motility,reduce the secretion of in-flammatory factor,thereby achieving the therapeutic effect of treating FD.
3.Advances in animal models of intestinal fibrosis
Xi WANG ; Hao LIU ; Xinyu SHEN ; Xin SUN ; Defang ZOU ; Renjun GU
Acta Laboratorium Animalis Scientia Sinica 2024;32(5):666-675
Intestinal fibrosis is a complication of inflammatory bowel disease,and its refractory and recurrent nature impose a serious disease burden on patients.The disease's pathogenes is not clear,and there is no effective treatment.Moreover,there is still a lack of recognized intestinal fibrosis models.In this paper,we review the method used to establish animal models of intestinal fibrosis both at home and abroad,and consider the clinical relevance,key characteristics,and advantages and disadvantages of the procedures.Intestinal fibrosis models were summarized according to the modeling period and method.
4.The association study of WWP2 gene polymorphisms with delayed encephalopathy after acute carbon monoxide poisoning
Fan ZHANG ; Jiao ZENG ; Wenqiang LI ; Yongkai HAN ; Ping ZHANG ; Xusheng WANG ; Jiapeng GU ; Renjun GU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2020;38(7):485-489
Objective:To investigate the association of WWP2 single nucleotide polymorphism (rs3790088, rs4247109) with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) , and explore the influences of DEACMP genetic predisposition. Methods:From November 2006 to December 2017, 235 DEACMP cases and 429 acute carbon monoxide poisoning (ACMP) cases were selected. All ACMP patients were followed up for more than 90 days without DEACMP. The DNA in all blood samples were extracted with the blood Genome DNA Extraction Kit. The method of Sequenom Mass Array SNP technique was used to detect the genotype and allele of WWP2. All DEACMP patients were assessed every 3 days after hospitalization by the Hasegawa Dementia Scale (HDS) and Activity of Daily Living Scale (ADL) . The distribution of genotypes in conformty with Hardy-Weinderg law was analyzed by goodness-of-fit χ 2 test, and χ 2 test was used for association analysis. Results:For rs3790088, there were 226 DEACMP cases and 414 ACMP cases. For rs4247109, there were 234 DEACMP cases and 428 ACMP cases. For rs3790088 and rs4247109 in WWP2 gene: there were not significant differences in the gene genotype distribution and allele frequency of both DEACMP group and ACMP group ( P>0.05) . According to gender, there were not significant differences in WWP2 gene genotype distribution and allele frequency between two female groups and two male groups ( P>0.05) . After analysis by genetic model, the genotype distributions in both DEACMP group and ACMP group were not significantly differences in three genetic models (codominant genetic model, recessive genetic model and dominant genetic model, P>0.05) . Conclusion:It has not confirmed the genetic correlation between the two gene single nucleotide polymorphisms (rs3790088, rs4247109) of WWP2 gene and the incidence of DEACMP.
5.The association study of WWP2 gene polymorphisms with delayed encephalopathy after acute carbon monoxide poisoning
Fan ZHANG ; Jiao ZENG ; Wenqiang LI ; Yongkai HAN ; Ping ZHANG ; Xusheng WANG ; Jiapeng GU ; Renjun GU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2020;38(7):485-489
Objective:To investigate the association of WWP2 single nucleotide polymorphism (rs3790088, rs4247109) with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) , and explore the influences of DEACMP genetic predisposition. Methods:From November 2006 to December 2017, 235 DEACMP cases and 429 acute carbon monoxide poisoning (ACMP) cases were selected. All ACMP patients were followed up for more than 90 days without DEACMP. The DNA in all blood samples were extracted with the blood Genome DNA Extraction Kit. The method of Sequenom Mass Array SNP technique was used to detect the genotype and allele of WWP2. All DEACMP patients were assessed every 3 days after hospitalization by the Hasegawa Dementia Scale (HDS) and Activity of Daily Living Scale (ADL) . The distribution of genotypes in conformty with Hardy-Weinderg law was analyzed by goodness-of-fit χ 2 test, and χ 2 test was used for association analysis. Results:For rs3790088, there were 226 DEACMP cases and 414 ACMP cases. For rs4247109, there were 234 DEACMP cases and 428 ACMP cases. For rs3790088 and rs4247109 in WWP2 gene: there were not significant differences in the gene genotype distribution and allele frequency of both DEACMP group and ACMP group ( P>0.05) . According to gender, there were not significant differences in WWP2 gene genotype distribution and allele frequency between two female groups and two male groups ( P>0.05) . After analysis by genetic model, the genotype distributions in both DEACMP group and ACMP group were not significantly differences in three genetic models (codominant genetic model, recessive genetic model and dominant genetic model, P>0.05) . Conclusion:It has not confirmed the genetic correlation between the two gene single nucleotide polymorphisms (rs3790088, rs4247109) of WWP2 gene and the incidence of DEACMP.
6.Effects of sleep epileptiform discharge on aural pre-attentive processing of children with benign childhood epilepsy with central-temporal spikes
Haorui DU ; Shuanrong LI ; Junlin MU ; Liuyi LI ; Zhaohui ZHANG ; Ning ZHANG ; Renjun GU
Chinese Journal of Applied Clinical Pediatrics 2017;32(13):1004-1007
Objective To explore the relationship between aural pre-attentive processing and non-rapid eye movement(NREM) sleep interictal epileptiform discharge (IED) in children with benign childhood epilepsy with central-temporal spikes (BECTS),in order to provide objective electrophysiological basis for early assessment of cognitive function lesion of BECTS children and intervention.Methods Twenty-nine children diagnosed as BECTS in the Second Affiliated Hospital of Xinxiang Medical University from February 2012 to November 2015 were selected,including 17 males and 12 females,and they were 4-14 years old with average age of (9.17 ±2.42) years,and the course of disease was 0.5-4.0 years.Twenty healthy children who had hospital routine check-up were selected as healthy control group,containing 14 males and 6 females,and they were 5-13 years old with average age of (8.55 ±3.09) years.Mismatch negativity (MMN) test was carried out in both groups,and the MMN amplitudes and latencies were analyzed.The long-term video electroencephalogram (VEEG) monitoring was conducted in the BECTS group to analyze the discharge phases (waking,sleep),locations and the IED indexes.The intergroup MMN indexes and the correlation between MMN and VEEG in the BECTS group were compared.Results The VEEG showed that the sleep structure of BECTS children did not change obviously.All children's epileptiform discharges were located in the Rolandic area,including 11 cases on the left side,8 cases on the right side,and 10 cases on both sides.Epilepsy discharge time:17 patients showed epileptiform discharges in sleep stages only and 12 patients showed epileptiform discharge in both sleep and waking stages.Epileptiform discharges of the 12 patients increased more significantly in sleep stage than thatin waking stage [(40.24 ± 25.15) times/min vs.(1.92 ± 1.38) times/rmin],and the difference was statistically significant (t =5.270,P =0.000).The sample IED index in stage 1 and 2 of NREM was (40.73 ± 10.69) times/min,in which 2 cases had IED indexes < 5 times/min,the IED indexes of 3 cases > 80 times/min who reached electrical status epilepticus during sleep,and others were 17.2-64.6 times/min.Clinical seizures in sleep stages were monitored in only 4 cases,which showed an electro-clinical segregation phenomenon from large quantities of sleep IED.The MMN amplitude was lower in the BECTS group than that in the healthy control group [(6.06 ± 1.89) μV vs.(7.28 ±1.64) μV],and the difference was statistically significant (t =2.346,P =0.025).Latency of BECTS group was longer than that in the healthy control group [(191.37 ±40.13) ms vs.(179.35 ±39.80) ms],but the difference was not statistically significant(t =1.037,P =0.355).Correlation analysis showed that the MMN amplitude was negatively related to discharge phases (r =-0.407,P < 0.05) and the IED indexes (r =-0.644,P < 0.01),and latency was positively related to the IED indexes (r =0.386,P < 0.05),while the other VEEG indexes were not correlated with MMN.Conclusions The BECTS children have aural pre-attentive processing disturbance,especially unconscious automatic processing ability impairment,which weakenattention switch action of automatic processing switching to focused attention processing and fail to filter irrelevant information effectively.Sleep IED is an important factor in impairing the formation and reinforcement of acoustic discrimination and memory trace,interfering the acquisition,processing,storage and matching of new information,which induces children pre-attentive processing disturbance.The MMN may discover the electrophysiological changes of children pre-attentive processing disturbance in early stage.
7.Relationship between changes of serum soluble Fas levels and delayed encephalopathy after acute carbon monoxide poisoning.
Ping ZHANG ; Yongkai HAN ; Guoyou ZHAO ; Jiapeng GU ; Ruihong WEI ; Renjun GU ; Chuansheng WANG
Journal of Southern Medical University 2013;33(1):150-151
OBJECTIVEBy explore the role of serum soluble Fas (sFas) in occurrence and progression of delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).
METHODSEnzyme-linked immunosorbent assay (ELISA) was used to detect serum sFas levels in 40 patients with DEACMP in acute stage and convalescent stage, with 36 healthy elderly subjects as the control group.
RESULTSSerum sFas levels of the patients with DEACMP in both the acute and convalescent stages showed no significant difference from those in the control group (P=0.737 and 0.137, respectively), nor was any significant difference found between the patients in acute and exacerbation stages (P=0.059).
CONCLUSIONSerum sFas is not involved in the occurrence and progression of DEACMP.
Adult ; Aged ; Aged, 80 and over ; Brain Diseases ; etiology ; Carbon Monoxide Poisoning ; blood ; complications ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; fas Receptor ; blood
8.Serum S100B protein and GFAP levels and their clinical significance in patients with delayed encephalopathy after acute carbon monoxide poisoning
Guohui HAN ; Renjun GU ; Wenqiang LI ; Ping ZHANG ; Fan ZHANG ; Tianyuan SHI ; Wei LI ; Xiahong WANG
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(12):1107-1110
Objective To explore the dynamic changes of serum S100B and glial fibrillary acidic protein (GFAP) levels and their clinical significance in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).Methods By means of enzyme-linked immunno-sorbent assay (ELISA),the serum S100B and GFAP levels from 33 DEACMP patients were assayed,and the condition changes were analyzed with three types of scale:the activity of daily living scale ( ADL),information-memory-concentration test (IMCT) and Hasegawa' s dementia scale(HDS).The comparison with 32 patients of acute carbon monoxide poisoning without DEACMP was also conducted.Results (1) The serum S100B( (0.60 ±0.21)ng/ml) and GFAP( (226.58 ±90.05 )ng/ml) in DEACMP group at acute stage were significantly higher than those in acute-CO-poisoning group ( (0.50 ± 0.20) ng/ml,( 183.04 ± 73.01 ) ng/ml) and those in DEACMP group at convalescent stage ( (0.51 ±0.16) ng/ml,(183.25 ±81.76)ng/ml) (all P values <0.05).(2)In DEACMP group,the serum S100B and GFAP at acute and convalescent stages were significantly correlated (at acute stage:r=0.466,P=0.006; at convalescent stage:r=0.365,P=0.037 ).(3)The S100B and GFAP in ineffective DEACMP patients at acute stage were significantly higher than those in the other groups ( all P values < 0.05 ).(4) In DEACMP group,the ADL,HDS and IMCT scores( (45.21 ± 9.69),(8.26 ± 6.31 ),(9.91 ± 7.52) ) at acute stage were significantly different from those at convalescent stages( (33.67 ± 13.62),( 15.91 ± 10.83),( 19.06 ± 10.37 ) ) ( all P values <0.01 ).Conclusion There is secondary brain insult (SBI) in DEACMP; glial activation may play an important role.The S100B and GFAP levels may be associated with the prognosis of DEACMP.
9.The value of brainstem auditory evoked potential and lower limb short latency somatosensory evoked potential for predicting delayed encephalopathy after acute carbon monoxide poisoning
Liuyi LI ; Junlin MU ; Renjun GU ; Ning ZHANG ; Haorui DU ; Wentao JIANG
Chinese Journal of Physical Medicine and Rehabilitation 2010;32(5):347-350
Objective To observe the changes of brainstem auditory evoked potential (BAEP) and lower limb short latency somatosensory evoked potential ( SLSEP) in acute carbon monoxide poisoning patients and explore the predictive value of BAEP and SLSEP for delayed encephalopathy after acute carbon monoxide poisoning (DEACMP). Methods Sixty-six acute CO poisoning patients was divided into a DEACMP group and a non-DEACMP group according to their symptoms. At the same time, 60 healthy subjects were included in the control group. BAEP and lower limb SLSEP dynamic testing were conducted at different time points. Results The abnormal rates in the first test of BAEP and lower limb SLSEP in the acute CO poisoning patients were 31.8% and 68.2% respectively.The BAEP results in the acute stage and later in the DEACMP group were comparable to those of the non-DEACMP group and the normal control group. Interpeak latency of Ⅰ-Ⅲ was not significantly different, but the remaining indices were significantly longer. Compared to the control group, the lower limb SLSEP readings in the acute DEACMP group and the non-DEACMP group were significantly longer. Compared to the non-DEACMP group and the control group, the SLSEP results of the DEACMP group were significantly extended. The specificity index of the BAEP predicted DEACMP up to 85.7% , significantly more than SLSEP testing. Conclusions Early detection of BAEP can help predict DEACMP. This finding should be promulgated and applied.
10.The changes of soluble interleukin-2 receptor in serum and cerebrospinal fluid of patients with delayed encephalopathy after acute carbon monoxide poisoning.
Renjun GU ; Xiuming ZHANG ; Luxian LU ; Hong LU ; Jinggui SONG ; Xinsheng GUO ; Zhongxing ZHANG ; Maixian LIU
Chinese Journal of Industrial Hygiene and Occupational Diseases 2002;20(5):350-352
OBJECTIVETo explore the changes of soluble interleukin-2 receptor(sIL-2R) in serum and cerebrospinal fluid (CSF) of patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).
METHODSThere were 31 patients with DEACMP, 32 patients with other encephalopathy and 31 controls in this study. The levels of sIL-2R in serum and CSF were detected by ELISA.
RESULTSSerum sIL-2R in patients with DEACMP[(329.21 +/- 160.99)U/ml] was significantly higher than that in control[(115.67 +/- 89.58) U/ml, P < 0.05)], but not significantly different from that in the other encephalopathy group[(367.50 +/- 123.14) U/ml, P > 0.05)]. CSF sIL-2R in patients with DEACMP[(54.48 +/- 43.04) U/ml] measured a little before discharge was significantly lower than that in patients with the other encephalopathy[(110.24 +/- 76.56) U/ml, P < 0.05)], but not significantly different from that in the control group[(34.96 +/- 22.70)U/ml, P > 0.05)]. At the pre-discharged period, CSF sIL-2R in patients with DEACMP[(100.26 +/- 93.65) U/ml] was significantly higher than that at the early stage of hospitalization[(52.28 +/- 43.31) U/ml, P < 0.05)]. No significant difference in serum sIL-2R was found between early stage of hospitalization[(338.34 +/- 161.53) U/ml] and pre-discharge [(351.31 +/- 175.93) U/ml, P > 0.05)].
CONCLUSIONThe occurrence of DEACMP may be related with immunopathological damage. The sIL-2R levels in serum and CSF may give information about the state of immunological function of the patients with DEACMP and may contribute to determining the patient's condition and prognosis.
Brain Diseases ; cerebrospinal fluid ; immunology ; Carbon Monoxide Poisoning ; cerebrospinal fluid ; immunology ; Enzyme-Linked Immunosorbent Assay ; Hospitalization ; Humans ; Receptors, Interleukin-2 ; analysis ; blood

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