[Mechanism of Shenling Baizhu Powder on treatment of alcoholic liver disease by regulating TLR4/NLRP3 pathway].

This study aims to investigate the regulatory effects of Shenling Baizhu Powder(SBP) on cellular autophagy in alcoholic liver disease(ALD) and its intervention effect through the TLR4/NLRP3 pathway. A rat model of chronic ALD was established by gavage of spirits. An ALD cell model was established by stimulating BRL3A cells with alcohol. High-performance liquid chromatography(HPLC) was utilized for the compositional analysis of SBP. Liver tissue from ALD rats underwent hematoxylin-eosin(HE) and oil red O staining for pathological evaluation. Enzyme-linked immunosorbent assay(ELISA) was applied to quantify lipopolysaccharides(LPS), tumor necrosis factor-alpha(TNF-α), interleukin-1 beta(IL-1ß), and interleukin-18(IL-18) levels. Quantitative reverse transcription polymerase chain reaction(qRT-PCR) was conducted to evaluate the mRNA expression of myeloid differentiation factor 88(MyD88) and Toll-like receptor 4(TLR4). The effect of different drugs on BRL3A cell proliferation activity was assessed through CCK-8 analysis. Western blot analysis was performed to examine the protein expression of NOD-like receptor pyrin domain-containing 3(NLRP3), nuclear factor-kappa B P65(NF-κB P65), phosphorylated nuclear factor-kappa B P65(p-P65), caspase-1, P62, Beclin1, and microtubule-associated protein 1 light chain 3(LC3Ⅱ). The results showed that SBP effectively ameliorated hepatic lipid accumulation, reduced liver function, mitigated hepatic tissue inflammation, and reduced levels of LPS, TNF-α, IL-1ß, and IL-18. Moreover, SBP exhibited the capacity to modulate hepatic autophagy induced by prolonged alcohol intake through the TLR4/NLRP3 signaling pathway. This modulation resulted in decreased expression of LC3Ⅱ and Beclin1, an elevation in P62 expression, and the promotion of autolysosome formation. These research findings imply that SBP can substantially enhance liver function and mitigate lipid irregularities in the context of chronic ALD. It achieves this by regulating excessive autophagic responses caused by prolonged spirit consumption, primarily through the inhibition of the TLR4/NLRP3 pathway.

[Effect of electroacupuncture on expression of NF-κB and TNF-α in renal tissue of rats with acute cerebral infarction].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on motor function, serum Cystatin C (Cys C) content, and expressions of tumour necrosis factor-α (TNF-α) and nuclear factor-kappa B (NF-κB) in renal tissue of rats with acute cerebral infarction (ACI), so as to explore its underlying mechanisms in protecting renal tissue after ACI. METHODS: Seventy-two male SD rats were randomly divided into three groups: sham operation, model and EA groups which were further randomly allocated to 1 d, 3 d, 7 d and 14 d subgroups (n=6 per subgroup). The ACI model was established by occlusion of the middle cerebral artery (MCAO). Rats of the EA group received EA of "Neiguan" (PC6) and "Zusanli" (ST36) for 30 min, once daily for 1, 3, 7 and 14 days, respectively. The motor function and content of Cys C were determined on the 1st, 3rd, 7th and 14th day after ACI. The expressions of TNF-α and NF-κB were detected by immunohistochemistry. RESULTS: Compared with the sham operation group, the motor function scores and the content of Cys C increased significantly on the 1st, 3rd, 7th and 14th d (P<0.01), while the numbers of TNF-α and NF-κB positive cells of the model group increased significantly on the 3rd, 7th and 14th d (P<0.01). After EA treatment, the motor function scores and the content of Cys C on the 7th, and 14th d, and the numbers of TNF-α and NF-κB positive cells on the 3rd, 7th and 14th d obviously decreased (P<0.05). CONCLUSION: EA at PC6 and ST36 can improve motor function and alleviate renal injury in ACI rats, possibly by regulating the expression of TNF-α, NF-κB in renal tissue and Cys C in serum.

IRF9 Affects the TNF-Induced Phenotype of Rheumatoid-Arthritis Fibroblast-Like Synoviocytes via Regulation of the SIRT-1/NF-κB Signaling Pathway.

OBJECTIVE: To discuss how IRF9 affects the fibroblast-like synoviocytes (FLS) in TNF-induced rheumatoid arthritis (RA) via the SIRT-1/NF-κB signaling pathway. METHODS: RA-FLS were isolated and divided into control, sh-IRF9, TNF, TNF + sh-Ctrl, TNF + sh-IRF9, TNF + sh-SIRT1, and TNF + sh-IRF9 + sh-SIRT1 groups. Biological features of FLS were evaluated by MTT, wound healing, and Transwell assays, respectively. Cell apoptosis and cycle were assessed flow cytometrically. Inflammatory cytokines were determined through enzyme-linked immunosorbent assay (ELISA), while IRF9 expression and SIRT1/NF-κB signaling pathway activity were measured by Western blotting. RESULTS: TNF increased IRF9 expression as well as NF-κB signaling activity and down-regulated SIRT1 of RA-FLS. Silencing IRF9 resulted in up-regulation of SIRT1 and blocked NF-κB signaling, with significant decreases in TNF-induced cell viability, migration, and invasion, prominent enhancement in apoptosis and the proportion of cells in G0/G1 phase, but a decrease in the proportion of cells in S and G2/M phases, and reduced levels of inflammatory cytokines. However, these changes were totally abolished after silencing SIRT1, i.e., the IRF9 shRNA-induced inhibitory effect on the growth of RA-FLS was reversed. CONCLUSION: Silencing IRF9 curbs the activity of the NF-κB signaling pathway via up-regulating SIRT-1, to further suppress TNF-induced changes in the malignant features of RA-FLS, and the secretion of inflammatory cytokines, with the promoted apoptosis.

Association Between Polymorphisms of Metabolic Enzyme Genes and Chromosomal Damage in Benzene-Exposed Workers in China.

OBJECTIVE: To provide better understanding of genetic susceptibility for health risk among current benzene-exposed workers. METHODS: Four hundred sixty one benzene-exposed workers and 88 matched controls were recruited, and their benzene exposure doses were monitored. Associations between genetic susceptibility for polymorphisms of metabolic enzymes CYP2E1 and NQO1, and expression of cytokinesis-block micronucleus (MN) were investigated. RESULTS: Mean MN frequency in the exposed workers was significantly higher than that in the control group (P < 0.01). Individuals with the NQO1 CC genotype showed significantly higher MN frequencies than those with the TT genotype (P < 0.05) in either single- or multiple-factor analyses. Age was an effect modifier for elevated MN frequency, while sex, smoking, and alcohol consumption had no relationship. CONCLUSION: Exposure to low dose of benzene among current workers can still cause health risk, especially among those with the NQO1 CC genotype.

Costs, Trends, and Related Factors in Treating Lung Cancer Patients in 67 Hospitals in Guangxi, China.

Lung cancer is a common disease with high mortality in China. Recent economic advances have led to improved medical capabilities, while costs associated with treating this disease have increased. Such change contributes to a commonly held belief that healthcare costs are out of control. However, few studies have examined this issue. Here, we use 34,678 hospitalization summary reports from 67 Guangxi hospitals (period 2013-2016) to document costs, temporal trends, and associated factors. Findings from this study are surprising in that they debunk the myth of uncontrolled healthcare costs. In addition, results and experiences from Guangxi are informative for other comparable regions.

Assessment of potential factors associating with costs of hospitalizing cardiovascular diseases in 141 hospitals in Guangxi, China.

BACKGROUND: The rising cost of healthcare is of great concern in China, as evidenced by the media features negative reports almost daily. However there are only a few studies from well-developed cities, like Beijing or Shanghai, and little is known about healthcare costs in rest of the country. In this study, we use hospitalization summary reports (HSRs) from admitted cardiovascular diseases patients in Guangxi hospitals during 2013-2016, and we investigate temporal trends of healthcare costs and associated factors. METHODS: By generalized additive model, we compute temporal trends of cost per stay (CPS), cost per day (CPD) and others. We then use generalized linear models to assess which factors associate with CPS and CPD. FINDINGS: Using a total of 760,000 HSRs, we find that CPS appears to be stabilized around $1040 until the middle of year 2015, before exhibiting a downward trend. Similarly, CPD exhibits similar stable pattern. Meanwhile, surgery-specific CPS showed an increase in year 2013-2014, and then stabilized. Drug costs account for over 1/3 of CPS, but they are gradually declining. Costs associated with physicians' and nurses' services represent less than 5% of CPS. We found that age, sex, marital status, occupation and payment methods are significantly associated with CPS or CPD. Interestingly, we found no association between patient ethnicity and these costs. However, we did find that minority patients use more secondary hospitals than Han patients. INTERPRETATIONS: Healthcare costs in Guangxi are stable, contrary to the rise portrayed by Chinese mass media. Several factors can be associated with healthcare costs, and these may be useful for developing evidence-based policies. In particular, there is a need to encourage more Han patients to seek care in primary and secondary hospitals.

Benchmark Doses Based on Abnormality of WBC or Micronucleus Frequency in Benzene-Exposed Chinese Workers.

OBJECTIVES: The aim of the study was to calculate benchmark dose for chromosomal damage and reduced white blood cell (WBC) associated with exposure to benzene (BZ). METHODS: A group of 317 exposed workers and 102 controls were examined for WBC count and genotoxicity by micronucleus (MN) frequency. The cumulative exposure concentration of BZ was calculated by ambient air BZ concentration at worksites in conjunction with job type and associated service duration. RESULTS: MN frequency (P < 0.01) was higher and WBC count was lower (P < 0.01) in exposed workers on average than in the controls. MN frequency was a more sensitive than WBC; workers older than 30 were more susceptible to abnormal MN frequency and WBC count reduction than those younger than 30. CONCLUSIONS: Benchmark dose estimates indicated that BZ exposure at levels below the current occupational exposure standard can induce genotoxicity and hematotoxicity.

Are polymorphisms in metabolism protective or a risk for reduced white blood cell counts in a Chinese population with low occupational benzene exposures?

BACKGROUND: Genetic variations in metabolic enzyme genes may enhance hematotoxicity in benzene-exposed populations. OBJECTIVE: To investigate the association between polymorphisms of metabolism genes and white blood cells (WBCs). METHODS: Three hundred and eighty-five benzene-exposed workers and 220 unexposed indoor workers were recruited in China. We explored the relationship between metabolic enzymes polymorphisms [glutathione S-transferase T1/M1 (GSTT1/M1) null, glutathione S-transferase P1 (GSTP1)rs1695, Cytochrome P450 2E1 (CYP2E1) rs3813867, rs2031920, rs6413432, microsomal epoxide hydrolase (mEH) rs1051740, rs2234922] by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis and WBC. RESULTS: The exposed group had lower WBC counts (P<0·001) than the unexposed group. Increased susceptibility to hematotoxicity, as evidenced by lower WBC counts, was found in workers with null-GSTT1 (P = 0·045), null-GSTM1 (P = 0·030), rs2031920 (P = 0·020), and rs3813867 (P = 0·014) genotypes. White blood cell counts were also lower in workers with null-GSTT1 and null-GSTM after adjusting for age, gender, smoking, and alcohol consumption. CONCLUSION: Null-GSTT1 and null-GSTM1 genotypes and Cytochrome P4502E1 (CYP2E1: rs2031920, rs3813867) may support the hematotoxicity of benzene-exposed workers in China, and we can make use of it to select susceptible population.

Effect of polymorphic metabolizing genes on micronucleus frequencies among benzene-exposed shoe workers in China.

It is well-known that metabolism of benzene is required for the induction of toxicity and consequent health problems. Therefore, genetic variation in benzene (BZ) metabolism genes can influence health outcomes. However, large population studies are needed to provide more evidence for such relationship. We have conducted a large population investigation (385 BZ-exposed shoe workers and 197 matched healthy controls) on the association between inheritance of certain BZ metabolizing genes and the expression of micronuclei (MN). The latter was based on the cytokinesis-blocked MN assay. We analyzed the polymorphisms of GSTM1, GSTT1, GSTP1 (rs1695), CYP2E1 (rs3813867), CYP2E1 (rs2031920), CYP2E1 (rs6413432), mEH exon 3 (rs1051740), mEH exon 4 (rs2234922). Univariate Poisson regression analysis demonstrated that the BZ-exposed workers had significantly increased MN frequency compared with the controls (FR=1.84, 95% CI: 1.56-2.18; P<0.001), and showed a cumulative exposure dose-response relationship. The CYP2E1 rs3813867 mutant allele (CC+GC) (FR 1.15, 95% CI 1.02-1.29; P=0.020) and rs2031920 variant allele (CT+TT) (FR=1.23, 95% CI: 1.09-1.37, P<0.01) was associated with higher MN frequency significantly compared with the wild genotype separately. Furthermore, the MN frequency in rs2031920 variant allele (CT+TT) (FR=1.17, 95% CI: 1.04-1.31, P<0.01) was also higher than the wild genotype when the age, gender and cumulative exposure dose was adjusted in Poisson regression. In addition, the CYP2E1, however, GSTM1null, GSTT1null, GSTP1 rs1695, rs6413432, rs1051740 and rs2234922 polymorphisms showed no association with MN frequency. Our results indicate that two promoter polymorphisms in the CYP2E1 gene, especially the rs2031920 variant allele, were involved with the BZ-induction of MN and may contribute to risk of cancer among exposed workers.

[Polymorphism of XRCC1 and chromosome damage in workers occupationally exposed to benzene].

OBJECTIVE: To explore the relationship between the polymorphisms of DNA repair gene (XRCC1 194, 280 and 399) and the chromosomal damage induced by benzene. METHODS: The chromosomal damage of the peripheral lymphocytes in 459 workers occupationally exposed to benzene and 88 non-exposed controls were detected with cytokinesis-block micronucleus (CBMN) assay. PCR-RFLP technique was used to measure polymorphisms in XRCC1 194, 280 and 399. RESULTS: It was found that the MN frequency (2.12‰ ± 1.88‰) of the exposed group was significantly higher than that (1.19‰ ± 1.68‰) of the control group (P < 0.05), in the exposed group, the MN frequency (3.00‰ ± 2.76‰) of older workers (> 35 years) was significantly higher than that (2.02‰ ± 1.71‰) of younger workers (≤ 35 years) (P < 0.05). The effect of genetic polymorphisms of XRCC1 on CBMN was not found. The haplotypes AAA/BAA, AAB/AAB, ABA/ABA, ABB/ABB could associated with the increased frequencies of total micronucleus (P < 0.05). CONCLUSION: Benzene exposure could result in chromosome damage. Age of workers and diplotypes of XRCC1 could associated with chromosomal damage induced by benzene.

Protective effects of glutamine preconditioning on ischemia-reperfusion injury in rats.

BACKGROUND: Hepatic ischemia-reperfusion injury is a common phenomenon in hepatic surgical procedures and can result in further severe damage. This study aimed to investigate the protective effects of glutamine preconditioning on hepatic ischemia-reperfusion injury in rats and its dose-dependency. METHODS: Thirty-two healthy male Wistar rats were randomly divided into four groups (n=8 per group). One group received 0.9% NaCl (control) and the other three received glutamine (Gln groups) 4 hours before ischemia. The Gln groups were named GL, GM, and GH according to the glutamine dose. The liver was subjected to 1 hour of ischemia and 2 hours of reperfusion. Two hours later, the levels of alanine aminotransferase (ALT), intracellular free calcium (Ca2+), and activity of Na+/K+ adenosine triphosphatase (ATPase) and superoxide dismutase (SOD) were assessed, and liver tissue sections were examined under a microscope. RESULTS: The Gln and control groups differed in the concentration of intracellular free calcium (P<0.05), and the activity of Na+/K+ ATPase and SOD in the Gln groups was higher than in the control group (P<0.05). The ALT level was lower in the GM and GH groups than in the control group (P<0.05). The levels of Na+/K+ ATPase and SOD rose gradually with increasing glutamine dose (P<0.05), and the concentration of Ca2+ declined gradually with increasing glutamine dose (P<0.05). The degree of hepatocyte injury was milder in the Gln groups than in the control group. CONCLUSIONS: Glutamine preconditioning protected effectively against hepatic ischemia-reperfusion injury. These protective effects were related to the dose of glutamine and due to the reduction of intracellular calcium overload and the improvements in the activity of Na+/K+ ATPase and SOD.

Preconditioning and postconditioning reduce hepatic ischemia-reperfusion injury in rats.

BACKGROUND: Ischemia-reperfusion injury occurs when ischemic tissues or organs suffer from further functional and structural damage when their blood supply recovers. This study aimed to contrast the protective effects of ischemic preconditioning and ischemic postconditioning in hepatic ischemia-reperfusion injury in rats. METHODS: Thirty-two healthy male Wistar rats were randomly divided into four groups: sham-operated (SO), ischemia-reperfusion (IR), ischemic preconditioning (I-pre), and ischemic postconditioning (I-post). Blood samples and hepatic tissue were taken from all groups after the experiments. RESULTS: There were significant differences between the IR, I-pre and I-post groups in alanine aminotransferase and aspartate aminotransferase levels, NF-kappaB p65 expression, apoptosis index and superoxide dismutase activity in hepatic tissue. There were no significant differences between the I-pre and I-post groups. CONCLUSIONS: Ischemic postconditioning and ischemic preconditioning reduce hepatic ischemia-reperfusion injury, but in clinical practice the former is a more appropriate choice.

[Expressions of MMP-2 and COX-2 mRNA in bladder transitional cell carcinoma and their correlation].

OBJECTIVE: To determine the levels of MMP-2 and COX-2 mRNA in bladder transitional cell carcinoma tissues and explore their relationship. METHODS: We enrolled in this study 42 patients with bladder transitional cell carcinoma, including Ta-T1 (n = 18), T2-T4 (n = 24), G1 (n = 12), G2 (n = 19), G3 (n = 11), metastasis (n =26) and non-metastasis (n = 16). Another 5 cases of normal bladder tissues were taken as controls, and the levels of MMP-2 and COX-2 mRNA were detected by RT-PCR. RESULTS: The relative expressions of COX-2 mRNA were 1.038 +/- 0. 484 in Ta-T1, 1.489 +/- 0.584 in T2-T4, 0.920 +/- 0.442 in G1, 1.338 +/- 0.584 in G2 and 1.632 +/- 0.515 in G3, all significantly higher than that of the controls (0.460 +/- 0.224, P < 0.05). And the corresponding relative levels of MMP-2 mRNA were 1.107 +/- 0.384, 1.604 +/- 0.425, 0.971 +/- 0.370, 1.445 +/- 0.378 and 1.755 +/- 0.387, also significantly higher than that of the latter group (0.423 +/- 0.227, P < 0.05). The COX-2 and MMP-2 mRNA levels in the tumor tissues with and without metastasis were 1.591 +/- 0.455 vs 0.815 +/- 0.430 and 1.676 +/- 0.339 vs 0.927 +/- 0.228, (P < 0.01), respectively, with a positive correlation between the mRNA level of COX-2 and that of MMP-2 (r = 0. 703, P < 0.01). CONCLUSION: MMP-2 and COX-2 mRNA are highly expressed in bladder transitional cell carcinoma tissues and their expressions are positively correlated with the degree of malignancy. MMP-2 and COX-2 might play a synergetic role in the pathogenesis and progression of bladder transitional cell carcinoma.