1.The Effect of Shenqi Jianxin Formula (参芪健心方) on Cardiomyocyte Pyroptosis in Chronic Heart Failure Model Rats Based on the NLRP3/Caspase-1 Signaling Pathway
Guoqing LIANG ; Ran XIA ; Yinyan WANG ; Pan LIU ; Jun ZHANG ; Xiaohua DAI
Journal of Traditional Chinese Medicine 2024;65(1):94-102
ObjectiveTo investigate the possible mechanism of Shenqi Jianxin Formula (参芪健心方) in the treatment of chronic heart failure (CHF) from the perspective of pyroptosis. MethodsFifty-two rats were randomly divided into sham operation group (n=8) and modeling group (n=44). In the modeling group, the anterior descending branch of the left coronary artery was ligated to construct CHF rat model. Forty successfully-modelled rats were randomly divided into model group, Entresto group, Shenqi Jianxin Formula group, MCC950 group and the combination group (Shenqi Jianxin Formula plus MCC950), with 8 rats in each group. In Shenqi Jianxin Formula group, 7.4 g/(kg·d) of Shenqi Jianxin Formula was given by gavage, while in Entresto group, 68 mg/(kg·d) of Entresto suspension was given by gavage; in MCC950 group, MCC950 was injected intraperitoneally with 10 mg/kg once every other day, and in the combination group, 7.4 g/(kg·d) of Shenqi Jianxin Formula was given by gavage, and MCC950 was injected intraperitoneally with 10 mg/kg once every other day; 10 ml/(kg·d) of saline was given by gavage in the sham operation group and the model group. After 3 weeks of continuous intervention, serum brain B-type natriuretic peptide (BNP), creatine kinase isoenzyme MB (CK-MB), interleukin 1β (IL-1β), and interleukin 18 (IL-18) levels were detected by ELISA; HE staining and MASSON staining were used to observe pathological changes in rat myocardium. Except for the Entresto group, western blot technique was used to detect the expression of NOD-like receptor protein 3 (NLRP3), caspase-1, and apoptosis-associated speck-like protein possessing a caspase-recruiting domain (ASC); RT-PCR was used to detect the expression of NLRP3 and caspase-1 mRNA. ResultsCompared with the sham operation group, HE staining of rats in the model group showed obvious myocardial injury, while MASSON staining showed increased area of collagen fibrosis, and serum BNP, CK-MB, IL-1β, IL-18, myocardial tissue NLRP3, caspase-1, ASC protein expression and NLRP3, caspase-1 mRNA expression were all elevated (P<0.05). Compared with those in the model group, cardiomyocyte injury of rats and collagen fibrosis area were reduced, and serum BNP, CK-MB, IL-1β, and IL-18 contents were all reduced in Shenqi Jianxin Formula group, Entresto group, MCC950 group, and the combination group; except for Entresto group, myocardial tissue NLRP3, caspase-1, ASC protein expression and NLRP3, caspase-1 mRNA expression were reduced in the remaining three medication group (P<0.05). Compared with Shenqi Jianxin Formula group, the MCC950 group and the combination group showed decreased serum IL-1β and IL-18 content, collagen fibrosis area, myocardial tissue NLPR3, caspase-1 protein expression, and caspase-1 mRNA expression, and decreased ASC and NLRP3 mRNA expression was shown in the combination group (P<0.05). Compared with MCC950 group, collagen fibrosis area was reduced, and serum IL-18 content, NLRP3, caspase-1 mRNA expression were reduced in the combination group (P<0.05). ConclusionShenqi Jianxin Formula can effectively improve the myocardial injury and heart failure in rats with CHF, and its mechanism may be related to the inhibition of cardiomyocyte pyroptosis through NLPR3/Caspase-1 pathway to reduce the level of intramyocardial inflammation. The combined use of MCC950 with Shenqi Jianxin Formula could more effectively inhibite myocardial pyroptosis, with better therapeutic result than single use of each part.
2.Relationship between changes in serum SP-D and sVCAM-1 levels and postoperative fracture healing and short-term prognosis in patients with traumatic rib fractures
Feiyue ZHANG ; Longqiang RAN ; Yong LI ; Peng JIA ; Huan LUO ; Xiaohua WANG
International Journal of Laboratory Medicine 2024;45(11):1281-1284,1288
Objective To investigate the relationship between the changes of serum human surfactant pro-tein D(SP-D)and vascular cell adhesion molecule-1(sVCAM-1)levels and postoperative fracture healing and short-term prognosis in patients with traumatic rib fractures.Methods A total of 102 patients with traumatic rib fractures who were treated in Banan Hospital Affiliated to Chongqing Medical University from January 2020 to December 2021 were selected as the research objects.After 8 months of treatment,the healing status of the patients was analyzed and divided into non-healing group(21 cases)and healing group(81 cases).Ac-cording to the pulmonary contusion,the patients were divided into pulmonary contusion group(19 cases)and non-pulmonary contusion group(83 cases).The serum levels of SP-D and sVCAM-1 were compared between the healing group and the non-healing group,and between the pulmonary contusion group and the non-pulmo-nary contusion group.The relationship between the changes of serum SP-D and sVCAM-1 levels and postop-erative fracture healing and short-term prognosis in patients with traumatic rib fractures were studied.Results The levels of SP-D and sVCAM-1 in the healing group were lower than those in the non-healing group(P<0.05).The levels of SP-D and sVCAM-1 in the non-pulmonary contusion group were lower than those in the pulmonary contusion group(P<0.05).Serum SP-D and sVCAM-1 levels were independent risk factors for postoperative fracture healing and short-term prognosis.Conclusion The changes of serum SP-D and sVCAM-1 levels in patients with traumatic rib fractures are related to fracture healing and short-term prognosis,which can be used as an important reference for prognosis evaluation.
3.Selection of optimal antibody titer and clinical value of passive agglutination for the diagnosis of Mycoplasma pneumoniae infection
Liangyu WANG ; Ran WEI ; Hongbin ZHU ; Dawei SHI ; Xiaohua HAN ; Lihua NING ; Deli XIN
Chinese Journal of Applied Clinical Pediatrics 2024;39(12):927-930
Objective:To investigate the optimal serum antibody titer in acute stage for the diagnosis of Mycoplasma pneumoniae (MP) infection by passive agglutination, and to evaluate the clinical diagnostic value of different antibody titers.Methods:A cross-sectional study.Eighty-eight pairs of clinical serum samples were collected from children with MP infection treated at the Department of Pediatrics in Shengjing Hospital of China Medical University from December 2016 to February 2017 and Children′s Hospital of Baotou in November 2019.The four-fold change of the double serum specific antibody titer was used as the gold standard, and the receiver operating characteristic (ROC) curve was plotted.When detecting the single serum in acute stage, different antibody titers were used as positive criteria to evaluate their clinical application value in the diagnosis of MP infection and find the most appropriate serum antibody titer as the diagnostic cut-off value.Results:(1)When the serum specific antibody titer ≥1∶40 was used as the positive criterion, the sensitivity was 72.9%, the area under the ROC curve was 0.817, and the specificity was 87.5%, which might cause overdiagnosis.When the serum specific antibody titer ≥1∶160 was used as the positive criterion, the specificity was 97.5%, the area under the ROC curve was 0.775, and the sensitivity was 52.1%, which might cause missed diagnosis.When the serum specific antibody titer ≥1∶80 was used as the positive criterion, the sensitivity was 60.4%, the specificity was 97.5%, and the area under the ROC curve was 0.823, overall performing better compared with the said two criteria.(2)After the disease lasted at least 5 days, blood samples were collected.About 72.5% of the children had antibodies, and 60.0% of the children had antibody titers ≥1∶80.Conclusions:(1)When the passive agglutination method is used to detect MP infection, antibody titer ≥1∶80 is recommended as the diagnostic standard.However, in clinical practice, the diagnosis of MP infection depends on clinical and other laboratory test results.(2) It is appropriate to collect blood samples on 5-7 days of illness.If MP infection is clinically suspected, and an antibody titer of 1∶40 is also suggestive, it can perform cooperative diagnosis based on molecular biology lab results or retest at a shorter interval.
4.Common bacteria in oropharynx of children with Mycoplasma pneumoniae pneumonia and its clinical significance
Lina HAN ; Dongxue ZHU ; Huisheng YAO ; Dongxing GUO ; Liangyu WANG ; Ran WEI ; Deli XIN ; Xiaohua HAN
Chinese Journal of Applied Clinical Pediatrics 2022;37(12):903-908
Objective:To investigate the common bacteria in the oropharynx of children with Mycoplasma pneumoniae pneumonia (MPP) and its clinical significance.Methods:A total of 134 children with MPP who were hospitalized in the Department of Pediatric Respiratory, Shengjing Hospital of China Medical University from December 2016 to June 2017 were selected as the research subjects, and 42 healthy children in the same hospital were selected retrospectively as the healthy control group during the same period.Fluorescent quantitative polymerase chain reaction Taqman probe was used to detect common oropharyngeal bacteria[ Streptococcus pneumoniae(SP), Moraxella catarrhalis(CTA), Haemophilus influenza(HI)] for the enrolled children.Firstly, the bacterial detection rate of MPP children and healthy children was compared.Then, according to age(<1 years old, 1-<3 years old, 3-<6 years old and 6-14 years old), bacterial detection[Mycoplasma pneumoniae(MP), MP+ bacteria]and bacterial species(MP+ SP, MP+ CTA, MP+ HI), 134 children with MPP were divided into groups to compare.Moreover, the relevant clinical datas were retrospectively analyzed by rank sum test and chi- square test. Results:Among 134 children with MPP, 79 (58.96%) children were detected bacteria, and 17 (40.48%) children were detected bacteria among 42 healthy children, with statistically significant differences( χ2=4.404, P<0.05). Compared with the MP group, the level of white blood cell (WBC)[8.5(6.7, 12.0)×10 9/L vs.7.8(5.8, 9.3)×10 9/L, Z=-2.232], C reactive protein(CRP)[19.2(7.2, 35.0) mg/L vs.8.4(3.4, 24.6) mg/L, Z=-2.810], lactate dehydrogenase(LDH)[286(244, 365) U/L vs.250(210, 302) U/L, Z=-2.474] and the incidence of lobar pneumonia[40.51%(32/79 cases) vs.18.18%(10/55 cases), χ2=7.510], pleural effusion[13.92%(11/79 cases) vs.3.64%(2/55 cases), χ2=3.917], refractory Mycoplasma pneumoniae pneumonia (RMPP)[34.18%(27/79 cases) vs.18.18%(10/55 cases), χ2=4.151] in MP+ bacteria group were higher; the course of fever[10(7, 12) d vs.8(6, 10) d, Z=-2.706] and duration of antibiotic use[16(13, 19) d vs.12(9, 16) d, Z=-3.747] in MP+ bacteria group were longer (all P<0.05). The level of WBC in MP+ SP group[12.20(7.80, 17.30)×10 9/L] was higher than that in MP+ HI group [6.75(5.37, 9.44)×10 9/L], and the differences were statistically significant( Z=11.574, P<0.05), and the incidence of lobar pneumonia in MP+ SP group [56.67%(17/30 cases)]was higher than that in MP+ CTA group [0(0/3 cases)]and MP+ HI group[18.75%(3/16 cases)], and the differences were statistically significant( χ2=9.770, P<0.05). Conclusions:Bacterial colonization or infection is more likely to occur in the oropharynx of children with MPP.When WBC, CRP, and LDH are significantly increased and the image shows a large consolidation or pleural effusion, it may indicate mixed bacterial infection, longer course of fever and higher incidence of RMPP, and the common mixed bacteria is SP.
5.Clinical evaluation of laboratory diagnosis of Mycoplasma pneumoniae
Liangyu WANG ; Xiaohua HAN ; Ran WEI ; Lina HAN ; Xijie LIU ; Jingyi LI ; Haiwei DOU ; Zhaoyong WU ; Shaogang LI ; Deli XIN ; Kunling SHEN
Chinese Journal of Applied Clinical Pediatrics 2022;37(16):1252-1256
Objective:To compare the laboratory diagnostic methods of Mycoplasma pneumonia(MP) and evaluate its clinical value.Methods:A prospective study.Throat swabs and double sera of children with MP infection were collected from December 2016 to January 2017 in Shengjing Hospital Affiliated to China Medical University; throat swab samples of healthy children aged 3 to 5 in Chaoyang District, Beijing were collected from March to May 2017.Passive agglutination (PA) was used to detect the double serum.Taking the 4-fold increase or decrease of the specific antibody titer of the double serum as the gold standard, the receiver operating characteristic curve (ROC) was drawn, and the laboratory methods for detecting MP infection were compared and evaluated.Results:(1)A total of 93 children with MP infection were clinically diagnosed, including 42 males (45.2%) and 51 females (54.8%), with an average age of 5.5 years.Sixty cases (64.5%) of MP infection were diagnosed.There were 349 healthy children, 198 males and 151 females, with an average age of 4.3 years.The positive rate of throat swab culture was 0.6% (2 cases), and the positive rate of fluorescent quantitative PCR(qPCR) was 18.9% (66 cases). (2) The culture specificity was the highest (100.0%) and the sensitivity was the lowest (65.0%). PA and enzyme linked immunosorbent assay (ELISA) were used to detect a single serum in the acute phase, the sensitivity was 71.7% and 86.5% respectively.ROC curve suggested that the current clinical diagnostic threshold MP specific antibody IgM ≥ 1∶160 was not the best diagnostic threshold.Molecular biological diagnostic methods were the most sensitive, RNA simultaneous and testing (SAT) was 85.0% and qPCR was 93.0%; while the specificity was low, 75.7% (SAT) and 63.6% (qPCR), respectively.(3) At the same time, MP nucleic acid (SAT, PCR) of throat swabs and a single serum (ELISA, PA) of children in acute phase were detected, the sensitivity was increased to 95.0%-100.0%, and the specificity was 63.6%-75.7%.Conclusions:Molecular biology is highly sensitive in diagnosing MP infection.It has asymptomatic infection or is carried after infection.Whether it needs treatment needs to be combined with clinical practice, when MP detection is positive.The detection of a single serum in the acute phase with a course of about 1 week has high sensitivity and is of reference value for the diagnosis of MP infection, but the diagnosis needs to be combined with clinical practice.The sensitivity and accuracy of detecting MP infection by single serological test combined with SAT in acute phase are higher than that by single application.
6.Preliminary study on the teaching mode of organ-system integrated curriculum based on motor system
Bao SU ; Xiaohua PENG ; Jianhua RAN ; Xiaoji LUO
Chinese Journal of Medical Education Research 2021;20(2):134-137
The traditional teaching mode of "subject-centered" has many shortcomings such as knowledge separation and lack of integrity. In order to make up for the deficiencies of this mode, the organ-system integrated medical teaching mode has gradually become the trend of medical teaching reform. Taking "motor system" as an example, this paper discusses the practice of the organ-system integrated medical teaching mode from four aspects: teaching implementation, teaching quality supervision, supporting assessment system and teaching effectiveness.
7. Preliminary clinical application of fluorescence microscopic imaging and computer-aided diagnosis system in the diagnosis of superficial cutaneous fungal infections
Jing TIAN ; Yue LIU ; Yehong MAO ; Xin LIAN ; Ning LI ; Xiaohua LYU ; Juan TAO ; Shaoqun ZENG ; Yi RAN ; Jingsi ZENG
Chinese Journal of Dermatology 2019;52(9):639-642
Objective:
To evaluate the accuracy of automated fluorescence microscopic imaging and computer-aided diagnosis system (AFMICADS) in the auxiliary diagnosis of superficial cutaneous fungal infections.
Methods:
Totally, 106 outpatients and inpatients with suspected superficial fungal infections were enrolled from clinical departments of Union Hospital, Tongji Medical College, Huazhong University of Science and Technology between July 2018 and September 2018. A total of 126 specimens were collected, including 83 skin scales and 43 nail parings. Each specimen was divided into 3 groups to be examined by conventional fungal microscopy, culture with modified Sabouraud dextrose agar and fluorescence microscopy (artificial fluorescence microscopy and AFMICADS-based fluorescence microscopy) respectively. A positive result was defined as that conventional fungal microscopy and/or fungal culture was positive. Consistency rate, sensitivity and specificity of the 3 microscopic methods were calculated. Statistical analysis was carried out with SPSS 10.0 software by using McNemar test and Kappa test for analyzing difference in the positive rate, as well as consistency, between the 3 microscopic methods and the positive standard, and by using efficiency test for comparing the consistency rate among the 3 microscopic methods.
Results:
Of 126 specimens, 124 (98.4%) were positive for artificial fluorescence microscopy, and 123 (97.6%) for AFMICADS-based fluorescence microscopy. Both positive rates of the above 2 microscopic methods were significantly higher than the positive rate of the positive standard (77.8%, both
8.Noninvasive prenatal screen of trisomy-21 using maternal plasma fetal free RNA allelic ratio
Yajuan XU ; Shanshan ZHAI ; Xiaohua LUO ; Yingying ZHANG ; Limin RAN ; Lidan REN
Chinese Journal of Obstetrics and Gynecology 2015;(8):568-575
Objective Through the detections of the heterozygote frequencies tests of fetal specific genes PLAC4 and COL6A2 mRNA alleles in plasma of pregnant women, to explore its possibility of application in the noninvasive prenatal screenings of trisomy-21. Methods A toltal of 500 cases (males and females 250 cases respectively)of Han ethnic groups with Henan Provice of China who were subject to the physical checkup clinic of the Third Affiliated Hospital,Zhengzhou University from June to December, 2013 were selected as the healthy physical checkup group, and such techniques as DNA sequencing and PCR-restriction fragment length polymorphism (RFLP) were adopted to the determinations of the heterozygote frequencies of the single nucleotide polymorphism(SNP)of the PLAC4 and COL6A2 genes in the maternal peripheral blood in the healthy physical checkup group, and the differential comparisons of the determination results of the SNP heterozygote frequencies and the corresponding heterozygote frequencies in the National Center for Biotechnology Information (NCBI) database;30 cases of healthy pregnant women who spontaneously underwent pregnancy checkups at the maternity clinic were randomly selected as the healthy pregnancy group, and real-time fluorescence quantitative reverse transcription-PCR technique was adopted for determining the expression levels of PLAC4 and COL6A2 mRNA in the peripheral blood of pregnant women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks;40 cases of the same phase were selected for acting as the specimens for the karyotype analyses of the amniotic fluid cells, among which 20 cases were trisomy-21, and the 20 cases of the negative control group, and reverse transcription-multiplex ligation dependent probe amplification (RT-MLPA) technique was adopted for screening the fetal trisomy-21. Results (1) The allele heterozygote frequencies of the SNP of the healthy physical checkup group:determinations of the genotypes and hybrid rates of the 10 SNP sites of the PLAC4 and COL6A2 genes indicated that those with higher heterozygote frequencies were respectively rs7717, rs559, rs1044598, rs59066201 and rs1042917, with population coverage of 98%. Among them, the allele hybrid rates of rs59066201 were never seen in the NCBI database;in the respective comparisons of the allele hybrid rates of rs8130833, rs9977003 and rs7844 with the hybrid rates of the NCBI database, the variations had statistical significance (P<0.05). (2) The expression levels of PLAC4 and COL6A2 mRNA of the different pregnancy weeks of the healthy pregnancy group: the levels of PLAC4 mRNA in the peripheral blood of women of 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks of pregnancy were respectively 7.22 ± 1.05, 8.02±1.41,9.51±1.69,11.33±2.11 and 13.31±2.58, with their expression levels rising along with the increase of the pregnancy weeks; among them, the comparison of pregnancy 8 weeks and pregnancy 10 weeks, the variations had no statistical significance (P>0.05);in the mutual comparisons among the expression levels of the various pregnancy weeks, the variations had statistical significance (P<0.05). The expression levels of COL6A2 mRNA in 8 weeks, 10 weeks, 12 weeks, 14 weeks and 16 weeks were respectively 8.95 ± 1.28, 11.19 ± 1.36,15.00 ± 1.58,16.87 ± 1.72 and 18.96 ± 2.79, with their expression levels rising along with the increase of the pregnancy weeks, and in the mutual comparisons between the expression levels of the various pregnancy weeks, the variations all had statistical significance (P<0.05). (3) Prenatal screenings of trisomy-21 in the validation group of the trisome:a total of 5 sites of rs7717, rs559, rs1044598, rs59066201 and rs1042917 were selected from the allele heterozygote frequencies of SNP sites were selected from the subjects of the healthy physical checkup group, and 10 cases of trisomy-21 specimens and 10 cases of negative CTR specimens were accurately determined, with the sensitivity reached 80%(17/20), and the specificity reached 90%(18/20). One case of the trisomy-21 and two negative cases were both homozygotes, and among the trisomy-21 specimens of two cases, only one SNP was a heterozygote, and it was impossible to conduct screenings on these 5 cases, with the screening accuracy reaching 100%(35/35). Conclusions Fetal specific genes PLAC4 and COL6A2 mRNA are expressed in the peripheral blood of pregnant women in different gestational age;its expression level increases with the increase of gestational age. Among them, five SNP including rs7717, rs559, rs1044598, rs59066201 and rs1042917 show highest heterogeneity rate, which is different from the corresponding heterogeneity rate in NCBI database. RT-MLPA technology is a rapid, effective, noninvasive and low cost method of prenatal screening 21 trisomy.
9.Serum levels of GP73 in patients with autoimmune liver disease
Qingling ZHAI ; Ran LIU ; Zhiqiang WANG ; Yubo HUANG ; Xiaohua HAO ; Xin FENG ; Hongshan WEI
Chinese Journal of Experimental and Clinical Virology 2014;28(4):268-270
Objective To evaluate the serum levels of GP73 for diagnosing autoimmune liver disease.Methods Three populations were included:80 patients with autoimmune liver disease; 80 patients with chronic hepatitis B,and 80 apparently healthy controls.Results The serum levels of GP73 in patients with autoimmune liver disease (112.3 ± 72.55 ng/ml) were significantly higher than those of patients with chronic hepatitis B (86.44 ± 60.69 ng/ml),and healthy controls (35.84 ± 11.8 ng/ml).However,there were no significant differences was observed in group of subjects with autoimmune hepatitis (107.4 ± 90.6 ng/ml),primary biliary cirrhosis ((89.0 ± 45.38 ng/ml),and primary sclerosing cholangitis (113.3 ± 50.87 ng/ml).Chosen the healthy control as "control group",the patients with chronic hepatitis B as "patients group ",the sensitivity and specificity of GP73 for diagnosing autoimmune liver disease were 75.0% and 97.53%,respectively.The area of ROC analysis was 0.93(95% CI;0.89-0.97).Conclusion The serum GP73 levels was significantly increased in patients with autoimmune liver disease,compared with those of healthy controls.For patients with higher levels of serum GP73,diagnosis of autoimmune liver disease should also be considered,especially for those patients with overlap-syndrome,except for chronic virus hepatitis and hepatocellular carcinoma.
10.The research about the correlation of colorectal adenocarcinoma gene fbxl 20 and E-cadherin
Xiaohua ZHU ; Shishan DENG ; Xingguo XIE ; Shiquan XU ; Maocheng RAN ; Shaoli XIE ; Jianjun ZHU
Practical Oncology Journal 2013;(6):509-515
Objective To study the relationship between the fbxl 20 gene、E-cadherin gene and the clinocopathologic features ,respectively , and the correlation about the fbxl 20 and the set gene .Methods The mRNA expression level of fbxl 20 and E-cadherin gene in 50 pairs of human colorectal adenocarcinoma tissues and matched normal tissues were detected by RT -PCR.Results The mRNA expression level of fbxl 20 gene in tumor tissues were significantly increased than that in normal tissues (0.479 ±0.141 vs.0.296 ±0.121,P=0.001),while the E-cadherin were decreased (0.440 ±0.026 vs.0.741 ±0.059,P=0.000).There was no found on the correlation between the mRNA expression level of fbxl 20 and the E-cadherin gene in the 50 tumor tissues(r=-0.165,P=0.251).However,the E-cadherin expressed level was negatively correlated with the fbxl 20 expressed level in the lower differentiation degree、in the high differentiation degree and the Duke′s D tumor tissues(r=-0.600,P=0.008;r=-0.784,P=0.017;r=-0.643,P=0.032).Conclusions The decreased mRNA expression level of E -cadherin in tumor tissues are largely due to the increased fbxl 20 gene expressed level ,which is related with the high mobility and invasion ability of the advanced tumor .

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