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Objective To explore the effect of Shuanglu Tongnao Formula on neuronal ferroptosis in ischemic stroke rats and its regulatory mechanism on the silent information regulator 2 homolog 1(SIRT1)/nuclear factor erythroid 2-related fac-tor 2(Nrf2)/glutathione peroxidase 4(GPx4)signaling pathways.Methods Twenty rats were selected as sham operation group by the random number table method,and the remaining seventy rats were made ischemic stroke rat models by the middle cerebral artery occlusion method.The rats that had been successfully modeled were randomly divided into the model control group,Shuanglu Tongnao formula group,Shuanglu Tongnao formula+SIRT1 inhibitor group(Shuanglu Tongnao formula+EX527 group),with 20 rats in each group.After 14 days,the rats were scored for neurological injury;TTC staining was applied to detect the area of cerebral infarction in rats;HE staining was applied to detect pathological changes in rat brain tissue;Nissl staining was applied to detect the number of neurons in rat brain tissue;the kit was applied to detect the levels of ferri ion(Fe2+),superoxide dismutase(SOD),glutathione(GSH),and malonaldehyde(MDA)in rat brain tissue;immunohistochemistry was applied to de-tect the positive expression of acyl-CoA synthetase long-chain family member 4(ACSL4),transferrin receptor(TFR),and ferritin heavy polypeptide 1(FTH1)proteins in rat brain tissue;Western blotting method was applied to detect the expression of SIRT1,Nrf2,GPx4,and cystine/glutamate antiporter solute carrier family 7 member 11(SLC7A11)proteins in rat brain tissue.Results Compared with the sham operation group,the neurological deficit score,cerebral infarction area,the contents of Fe2+and MDA,and the protein expressions of ACSL4 and TFR in model control group were increased(P<0.05);the number of neurons,the con-tents of SOD and GSH,the protein expression of FTH1,SIRT1,Nrf2,GPx4,and SLC7A11 were all reduced(P<0.05).Compared with the model control group,the neurological deficit score,cerebral infarction area,the contents of Fe2+and MDA,and the protein expression of ACSL4 and TFR in the Shuanglu Tongnao formula group were reduced(P<0.05),and the number of neurons,the contents of SOD and GSH,the protein expressions of FTH1,SIRT1,Nrf2,GPx4,and SLC7A11 are all increased(P<0.05).The results of the SIRT1 inhibitor supplementation experiment showed that the SIRT1 inhibitor reversed the inhibitory effect of Shuan-glu Tongnao formula on neuronal ferroptosis,while also inhibited the expression of Nrf2 and GPx4(P<0.05).Conclusion The Shuanglu Tongnao formula may inhibit neuronal ferroptosis in ischemic stroke rats by activating the SIRT1/Nrf2/GPx4 signa-ling pathway.
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ObjectiveTo quantitatively investigate the changes in the total volume and contour density of hepatic oval cells （HOC） in hepatic lobules of rats with carbon tetrachloride （CCl4）-induced hepatic fibrosis. MethodsA total of 11 healthy male Sprague-Dawley rats were randomly divided into control group with 5 rats and hepatic fibrosis group with 6 rats， and CCl4 and olive oil suspension were injected subcutaneously twice a week， 3 mL/kg each time. After five weeks of hepatic fibrosis modeling， five liver tissue blocks with a size of about 1 mm3 were randomly selected from the liver of each rat to prepare one Epon812 epoxy resin-embedded ultrathin section， and the stereological method and transmission electron microscopy were used for the quantitative analysis of the total volume and contour density of HOC in the hepatic lobules of rats. In addition， four liver tissue blocks with a thickness of 2 mm were randomly selected from the remaining liver of each rat to prepare two paraffin-embedded Masson staining sections， and the degree of liver fibrosis in each rat was qualitatively evaluated according to the Metavir staging criteria for liver fibrosis. The independent-samples t test was used for comparison of continuous data between groups. ResultsThe quantitative stereological analysis showed that the total volume of HOC in hepatic lobules was 15.40±7.63 mm3 in the control group and 146.80±114.00 mm3 in the liver fibrosis group， and compared with the control group， the total volume of HOC in hepatic lobules of rats in the liver fibrosis group was significantly increased by 8.53 times （t=-2.551， P=0.031）； the contour density of HOC in hepatic lobules was 56.20±40.40 in the control group and 566.50±317.00 in the liver fibrosis group， and compared with the control group， the contour density of HOC in hepatic lobules of rats in the liver fibrosis group was significantly increased by 9.08 times （t=-3.539， P=0.006）. Qualitative observation showed that liver fibrosis stage of rats reached stage Ⅱ-Ⅲ according to the Metavir scoring criteria， and massive proliferation of HOC was observed around the proliferation site of hepatic stellate cells in the perisinusoidal space of rats. ConclusionCCl4 induces significant proliferation of HOC in hepatic lobules of rats with liver fibrosis.
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Objective To investigate the expression levels of HBV pregenomic RNA (pgRNA) and hepatitis B core-related antigen (HBcrAg) in circulating serum of chronic hepatitis B (CHB) patients after withdrawal from nucleos(t)ide analogues (NUC), as well as the correlation of HBV pgRNA and HBcrAg levels in circulating blood in different periods of time with recurrence in CHB patients after drug withdrawal. Methods Among the patients who attended the outpatient service of Affiliated Hospital of North Sichuan Medical College from December 2019 to July 2022, a total of 108 CHB patients who received anti-HBV therapy for at least 5 years and met the criteria for drug withdrawal in 2017 EASL Guidelines were enrolled. According to the time of drug withdrawal, the patients were divided into 4-, 12-, and 24-week groups after drug withdrawal, and according to the presence or absence of recurrence, they were divided into recurrence group and non-recurrence group. Quantitative real-time PCR was used to measure the level of HBV pgRNA in circulating serum of CHB patients; ELISA was used to measure the expression level of HBcrAg in peripheral venous blood; quantitative real-time PCR was used to measure HBV DNA load with high accuracy. The t -test was used for comparison of continuous data between two groups. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. The Pearson correlation test was used to investigate the correlation between the indices in circulating blood. Results For the CHB patients after drug withdrawal, the recurrence rate was 17.1% at 4-12 weeks, cumulative recurrence rate reached 29.3% after 24 weeks of follow-up, the patients with positive HBV DNA alone accounted for 64.3% and 60.0%, respectively, those with positive HBeAg alone accounted for 28.5% and 20.0%, respectively, and those with positive HBV DNA and HBeAg accounted for 7.1% and 20.0%, respectively. The expression levels of HBV pgRNA, HBcrAg, and HBV DNA in circulating serum of CHB patients at 24 weeks after drug withdrawal were significantly higher than those at the time of drug withdrawal and at 4 weeks after drug withdrawal, and there was a significant difference between groups at different time points (all P 0.05). Conclusion The recurrence group had significantly higher expression levels of HBV pgRNA and HBcrAg than the non-recurrence group at the time of drug withdrawal, suggesting that the levels of HBV pgRNA and HBcrAg in the CHB patients of the non-recurrence group at the time of drug withdrawal may be used as the reference thresholds for safe drug withdrawal in CHB patients, and measurement of HBV pgRNA and HBcrAg may be one of the potential reference indicators for the selection of anti-HBV treatment endpoints in the future.
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Objective:To investigate the factors affecting prognosis of pancreatic cancer patients with carbohydrate antigen 19-9 (CA19-9) negative before surgery.Methods:Retrospectively analyzed the clinic pathological data of 158 patients with pancreatic cancer who underwent radical surgery between November 2011 and August 2022, including 84 males, and 74 females, aged (65±9) years old. Patients were divided into two groups according to CA 19-9 levels over a threshold of 37 U/ml. Patients with CA19-9 levels ≤37 U/ml were included in the CA19-9 negative group ( n=34), while patients with CA19-9 levels >37 U/ml were included in the CA19-9 positive group ( n=124). We performed a comparison between the two groups regarding demographic and clinical data, tumor staging, lymphovascular and nerve invasion and so on. Survival data were extracted from outpatient or telephone records during follow-up. The Kaplan-Meier method was used to draw the survival curve, and the survival rate was compared by log-rank test. Multivariate analysis was performed using the Cox regression analysis model. Results:CA19-9 expression was associated with tumor location and nerve invasion (both P<0.001). There was no significant difference in cumulative survival rate ( χ2=0.07, P=0.787) and progression-free survival rate ( χ2=0.13, P=0.721) between two groups. Multivariate analysis showed that lymphovascular invasion ( HR=5.962, 95% CI: 2.189-16.239, P<0.001) was a prognostic factor for overall survival in the CA19-9 negative group. Age ≤60 years old ( HR=0.306, 95% CI: 0.118-0.796, P=0.015) and lymphovascular invasion ( HR=3.096, 95% CI: 1.232-7.780, P=0.016) were prognostic factors for progression-free survival. Conclusion:Age ≤60 years old and lymphovascular invasion are the influencing factors of poor prognosis in pancreatic cancer patients with CA19-9 negative.
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Objective:To establish and validate a prognosis prediction model of gall bladder cancer after surgery based on the albumin-bilirubin (ALBI) grade.Methods:The clinicopathological data of 161 patients with gallbladder cancer undergoing radical surgery in the First Hospital of Jiaxing between March 2011 to January 2020 were retrospectively analyzed, including 44 males and 117 females, aged (65±9) years old. The patients were grouped by the levels of ALBI score. The survival data were obtained through telephone or outpatient review. The Kaplan-Meier method was used to draw the survival curve. Multivariate analysis was performed using the Cox regression analysis model. The time-dependent receiver operating characteristic (ROC) curve was plotted to compare the prognostic value of the scoring system. A nomogram based on ALBI grade was established and its predictive performance was evaluated.Results:The 1, 3, 5 years overall survival (OS) rates of patients with gallbladder cancer were 76.7%, 52.5%, and 41.9%, respectively. The time-dependent ROC curves revealed that the area under the curve (AUC) values of ALBI grade were 0.659, 0.597 and 0.599 for 1, 2 and 3 years, showing a good prognostic performance. Multivariate regression analysis showed that poorer tumor differentiation ( HR=2.890, 95% CI: 1.816-4.600, P<0.001), TNM Ⅲ/Ⅳ stage ( HR=2.832, 95% CI: 1.781-4.503, P<0.001), ALBI grade 2 ( HR=1.595, 95% CI: 1.017-2.500, P=0.042), and ALBI grade 3 ( HR=3.938, 95% CI: 1.375-11.278, P=0.011) were independent risk factors for OS. The nomogram established with the independent risk factors such as ALBI grade showed a good predictive value for OS. The ROC curve results showed that the AUC values for 1, 3 and 5 years were 0.796, 0.806 and 0.799, respectively. The calibration plots and clinical decision curve analysis (DCA) showed the clinical feasibility of this nomogram. Conclusion:The nomogram model based on ALBI grading has a good predictive value for gallbladder cancer after surgery, which could guide the prognosis and individualized treatment decision-making.
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Exosomes, as a type of multivesicular bodies secreted by various cells in human body, exist in many kinds of body fluids in the body. Exosomes have the structure of lipid bilayer membranes to protect the structure and function stability of their contents and they carry a large number of active substances including microRNA (miRNA) and mRNA. Studies have shown that exosomes and their miRNA are closely associated with the development and progression of liver fibrosis and regulate such process by acting on hepatic stellate cells to change their activation, proliferation, apoptosis, and migration. This article elaborates on the biological characteristics of exosome miRNA and its association with liver fibrosis, analyzes its important role in the development and progression, diagnosis and treatment, and prognostic evaluation of liver fibrosis, and points out that exosome miRNA may become a new potential target for the diagnosis, treatment, and prognostic evaluation of liver fibrosis.
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Objective To investigate whether emodin combined with 5AzA-cdR can enhance the demethylation of tumor suppressor genes p16,RASSF1A and ppENK in nude mice with subcutaneously transplanted pancreatic cancer.Methods Pancreatic cancer cells Panc1 burdened subcutaneous xenograft nude mice model was established,which were randomly divided into control group,emodin group,5AzA-cdR group and emodin combined 5AzA-cdR group (combined group).The growth of transplanted tumors wasobserved in each group.Methylation specific PCR (MSP) was used to detect the methylation levels of p16,RASSF1A and ppENK in the xenograft tumor tissue among three groups.The mRNA and protein expression of three tumor suppressor genes were detected by FQ-PCR and Western blotting,respectively.Results The weight of xenografts in the control group,emodin group,5AzA-cdR group,and combination group were (0.28 ±0.01),(0.17 ± 0.01),(0.12 ± 0.02),(0.08 ± 0.01)g,respectively.The tumor volume was (517 ±0.02),(382 ± 0.01),(232 ± 0.03),(169 ± 0.01) mm3.The methylation levels of p16 were 1.00 ± 0.00,0.89 ± 0.02,0.63 ± 0.02,and 0.19 ± 0.01;the methylation levels of RASSF1A were 1.00 ± 0.00,0.88 ± 0.02,0.51 ± 0.01,and 0.32 ± 0.01;the methylation degree of ppENK was 1.00 ± 0.00,0.92 ± 0.02,0.77 ± 0.02 and 0.31 ± 0.01,respectively.The expression of p16 mRNA was 1.00 ± 0.00,1.71 ±0.02,2.67 ± 0.02,3.81 ± 0.01.The expression of RASSF1A mRNA was 1.00 ± 0.00,1.92 ±0.02,2.73 ± 0.03,3.77 ± 0.01.The expression of ppENK mRNA was 1.00 ± 0.00,1.69 ± 0.03,2.17 ± 0.02 and 4.28 ± 0.01.The expression of p16 protein was 1.00 ± 0.00,1.71 ± 0.02,2.67 ± 0.02,3.81 ± 0.01;the expression of RASSF1A protein was 1.00 ± 0.00,1.92 ± 0.02,2.73 ± 0.03.3.77 ± 0.01;ppENK protein expression levels were 1.00 ±0.00,1.69 ±0.03,2.17 ±0.02,4.28 ±0.01.The weight and volume of xenografts in the three treatment groups were significantly smaller than those in the control group.The methylation of three tumor suppressor genes was lower than that of the control group,and the expression of tumor suppressor mRNA and protein was all significantly higher than the control group,which the combination drug group was also significantly stronger than that in emodin group and 5AzA-cdR group,and the differences were statistically significant (P ＜ 0.05 or ＜ 0.01).Conclusions The combination of emodin and 5AzA-cdR can enhance the demethylation effect of 5A6A-cdR on the tumor suppressor genes p16,RASSF1A and ppENK in the tumor tissue of pancreatic cancer xenograft model.
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Objective To explore the compound compatibility regulation of oral traditional Chinese medicines (TCM) for the prevention and treatment of infantile repeated respiratory infection.Methods The oral TCM compound literatures for the prevention and treatment of infantile repeated respiratory infection were collected from four databases,including China Biology Medicine disc,CNKI,VIP Periodical and Wanfanf Data.The retrieval time was from January 2006 to July 2006.Then the TCM oral compound prescriptions meeting the requirements were obtained by the literature arrangement and performed the terms standardized processing for extracting the information and establishing Excel table.Then the Kaiyuan software package R i386 3.3.0 was used to conduct the drug frequency,association rules analysis and clustering analysis.Results A total of 311 prescriptions were screened out and 208 kinds of traditional Chinese medicine were analyzed.It was found that the higher compatibility frequencies of TCM were atractylodes,radix glycyrrhizae,windproof,radix astragali,radix pseudostellariae,poria cocos,dried tangerine or orange peel,etc.;the strong association rules prompted that modified Yupinfeng Powder composed of core medicines such as astragalus,atractylodes and windproof was the basic formula for prevention and treatment of infantile repeated respiratory infection;the clusters analysis found some TCM societies such as emembranous Astragalus mongholicus-keel-oysters,Atractylodes-Poria cocos-licorice-Astragalus mongholicus-dangshen.Conclusion The drug frequency,association rules analysis and clustering analysis can reveal the compatibility rules of TCM prescriptions for preventing and treating infantile repeated respiratory to provide reference basis for optimizing the clinical prescription and improving the curative effect.
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Objective To investigate the clinical characteristics and risk factors for healthcare-associated infection (HAI)in patients with severe chronic hepatitis B (CHB),and provide theoretical basis for preventing and controlling HAI.Methods Retrospective survey was used to investigate the occurrence of HAI in hospitalized patients with severe CHB in a hospital between January 2012 and January 2015,risk factors for HAI were analyzed. Results A total of 126 patients with severe CHB were investigated,49 patients developed 106 times of HAI, incidence of HAI was 38.89%.The main HAI site was respiratory tract (n=47,44.34%),the next was abdominal cavity (n=34,32.08%).A total of 76 isolates of pathogens were detected,gram-negative bacteria,gram-positive bacteria,and fungi accounted for 53.95%(n =41 ),43.42%(n =33),and 2.63%(n =2)respectively.Risk factors for HAI in patients with severe CHB were patients ’ age ≥ 60 years, length of hospital stay ≥ 30 days, complications,invasive operation,serum albumin < 35 g/L,and white blood cell count (WBC)< 4 × 109/L. Conclusion Incidence of HAI in patients with severe CHB is high,the majority are respiratory tract and abdominal cavity infection,risk factors are old age,long length of hospital stay,invasive operation,hypoalbuminemia,and low WBC count.