Objective·To investigate the therapeutic effects of neferine(Nef)on intervertebral disc degeneration(IDD)and the underlying regulatory pathways.Methods·The effects of Nef on the viability and proliferation of nucleus pulposus cells were assessed using the cell counting kit-8(CCK-8)assay.Molecular docking software was employed to analyze the potential binding sites of Nef within the Kelch domain of kelch-like ECH-associated protein 1(KEAP1).Tumor necrosis factor-α(TNF-α)was used to induce ferroptosis and inflammation in nucleus pulposus cells.Western blotting was performed to detect the expression levels of nuclear factor erythroid 2-related factor 2/glutathione peroxidase 4(NRF2/GPX4)pathway-and nuclear factor-KB(NF-κB)pathway-related proteins under TNF-α stimulation with or without Nef.The effect of Nef on the metabolism of extracellular matrix in nucleus pulposus cells was evaluated using high-density cell culture.A needle puncture-induced IDD rat model was established,and 5 μL of 1.5 μmol/L Nef was injected twice into the intervertebral disc at the Co3/4 level(IDD+Nef group),while an equivalent volume of PBS was injected into the Co2/3 disc(IDD group).After 4 weeks,the intervertebral space height was detected by X-ray,disc degeneration was detected by magnetic resonance imaging,and disc structure was evaluated by histological staining.Results·The CCK-8 assay revealed that Nef at concentrations of 1.5 μmol/L and below did not inhibit the viability and proliferation of nucleus pulposus cells.Molecular docking results suggested that Nef might activate NRF2 by directly binding to the KEAP1 Kelch domain,thereby reducing the interaction between KEAP1 and NRF2.Western blotting indicated that Nef significantly increased the expression of the key ferroptosis-inhibiting proteins NRF2 and GPX4,while decreasing the expression of the phospho-P65 protein in the NF-κB pathway(all P＜0.05).The high-density culture of nucleus pulposus cells demonstrated that Nef mitigated the TNF-α-induced degradation of the extracellular matrix(P＜0.05).Animal study results showed that compared to the IDD group,the IDD+Nef group exhibited a greater intervertebral disc space height,a lower Pfirrmann grade(both P＜0.05),and a reduced degree of histological degeneration.Conclusion·Nef may inhibit TNF-α-induced ferroptosis in nucleus pulposus cells by activating the KEAP1/NRF2/GPX4 pathway and reduce TNF-α-induced inflammation and extracellular matrix degradation by suppressing the NF-κB pathway,thereby alleviating IDD in rats.

Objective·To investigate the therapeutic effects of neferine(Nef)on intervertebral disc degeneration(IDD)and the underlying regulatory pathways.Methods·The effects of Nef on the viability and proliferation of nucleus pulposus cells were assessed using the cell counting kit-8(CCK-8)assay.Molecular docking software was employed to analyze the potential binding sites of Nef within the Kelch domain of kelch-like ECH-associated protein 1(KEAP1).Tumor necrosis factor-α(TNF-α)was used to induce ferroptosis and inflammation in nucleus pulposus cells.Western blotting was performed to detect the expression levels of nuclear factor erythroid 2-related factor 2/glutathione peroxidase 4(NRF2/GPX4)pathway-and nuclear factor-KB(NF-κB)pathway-related proteins under TNF-α stimulation with or without Nef.The effect of Nef on the metabolism of extracellular matrix in nucleus pulposus cells was evaluated using high-density cell culture.A needle puncture-induced IDD rat model was established,and 5 μL of 1.5 μmol/L Nef was injected twice into the intervertebral disc at the Co3/4 level(IDD+Nef group),while an equivalent volume of PBS was injected into the Co2/3 disc(IDD group).After 4 weeks,the intervertebral space height was detected by X-ray,disc degeneration was detected by magnetic resonance imaging,and disc structure was evaluated by histological staining.Results·The CCK-8 assay revealed that Nef at concentrations of 1.5 μmol/L and below did not inhibit the viability and proliferation of nucleus pulposus cells.Molecular docking results suggested that Nef might activate NRF2 by directly binding to the KEAP1 Kelch domain,thereby reducing the interaction between KEAP1 and NRF2.Western blotting indicated that Nef significantly increased the expression of the key ferroptosis-inhibiting proteins NRF2 and GPX4,while decreasing the expression of the phospho-P65 protein in the NF-κB pathway(all P＜0.05).The high-density culture of nucleus pulposus cells demonstrated that Nef mitigated the TNF-α-induced degradation of the extracellular matrix(P＜0.05).Animal study results showed that compared to the IDD group,the IDD+Nef group exhibited a greater intervertebral disc space height,a lower Pfirrmann grade(both P＜0.05),and a reduced degree of histological degeneration.Conclusion·Nef may inhibit TNF-α-induced ferroptosis in nucleus pulposus cells by activating the KEAP1/NRF2/GPX4 pathway and reduce TNF-α-induced inflammation and extracellular matrix degradation by suppressing the NF-κB pathway,thereby alleviating IDD in rats.

Objective:To study the incidence and risk factors of central vein stenosis (CVS) in chronic kidney disease (CKD) patients who received arteriovenous fistula (AVF) creation for the first time, as well as effects of CVS on patency of ipsilateral AVF.Methods:It was a retrospective study. The CKD patients who received AVF creation for the first time in the First Affiliated Hospital of Zhengzhou University from January 2019 to August 2020, with central vein digital subtraction angiography (DSA) results prior to angioplasty were selected as the study subjects. The differences of incidence of CVS in CKD patients with/without a history of cervical catheterization and primary patency rates of AVF between CVS and non-CVS groups were compared. Logistic regression analysis method was applied to analyze the influencing factors of CVS in CKD patients. Kaplan-Meier method was used to analyze the primary patency rate of AVF. Cox regression analysis method was used to analyze the effect of CVS on the primary patency of ipsilateral AVF.Results:A total of 283 CKD patients aged (50.45±14.76) years were enrolled in the study, including 165 males (58.3%). The dialysis age was 0.5 (0, 7.0) months. There were 55 patients (19.4%) diagnosed with CVS before AVF, including 39 patients with stenosis <50% and 16 patients with stenosis ≥50%. The incidence of CVS in patients with history of right internal jugular vein central venous catheter insertion was significantly higher than that in those without this history [60.5% (26/43) vs. 9.9% (15/151), χ2=51.274, P<0.001]. Multivariate logistic regression analysis results showed that hemodialysis catheters indwelling time ≥3 months elevated the risk of CVS ( OR=4.345, 95% CI 1.540-12.263, P=0.006). A subset of 268 patients who had AVF creation ipsilateral to CVS were analyzed to determine the effects of CVS on patency of AVF. The median follow-up time was 34 months. The primary patency rate of AVF in the moderate to severe CVS group was significantly lower than that in the non-CVS group (5/7 vs. 58/228, χ2=7.720, P=0.005). The primary patency rates of AVF in the subclavian vein stenosis group and superior vena cava stenosis group were significantly lower than those in the brachiocephalic vein stenosis group (4/5 vs. 8/27, χ 2=6.974, P=0.008; 6/8 vs. 8/27, χ 2=6.908, P=0.009, respectively). Moderate to severe CVS and combined diabetes were independent influencing factors of primary patency of AVF ( HR=4.362, 95% CI 1.644-11.574, P=0.003; HR=2.682, 95% CI 1.624-4.431, P<0.001, respectively). Conclusions:The incidence of CVS is higher in CKD patients who establish an arteriovenous fistula for the first time. Hemodialysis catheter indwelling time ≥3 months is an independent risk factor of CVS. The moderate to severe CVS is an independent risk factor of primary patency of AVF.

Objective:To investigate the risk factors for catheter-related bloodstream infection (CRBSI) in hemodialysis (HD) patients with tunnel-cuffed catheter (TCC) and construct a risk prediction model for the prevention and treatment of catheter infection.Methods:It was a retrospective study. Patients who had their TCC removed in Hemodialysis Access Center of the First Affiliated Hospital of Zhengzhou University from July to December 2020 were randomly divided into a training set (for model building) and a validation set (for model validation) in the ratio of 7∶3. The training set was divided into CRBSI group and non-CRBSI group with reference to the 2019 Kidney Disease Outcomes Quality Initiative clinical practice guidelines for vascular access, and the risk factors for the occurrence of CRBSI were analyzed. The odds ratio ( OR) values of the variables in the multivariate logistic regression analysis were applied to construct a risk prediction model, and the assessment ability of the model was validated in the validation set. Results:A total of 254 HD patients were included. The training set consisted of 179 patients with male-to-female ratio of 1.36∶1, age of (55.81±15.95) years old, median dialysis age of 18(8, 27) months, median TCC retention time of 15(5, 24) months, and 40 patients with confirmed CRBSI. Logistic regression analysis showed that, combined diabetes ( OR=2.711, 95% CI 1.174-6.258, P=0.019), history of catheter-related infection within 3 months ( OR=3.674, 95% CI 1.541-8.760, P=0.003), more than 4 times nursing interventions within 1 month ( OR=3.128, 95% CI 1.343-7.283, P=0.008), and central venous disease ( OR=2.572, 95% CI 1.130-5.854, P=0.024) were the independent influencing factors for CRBSI occurrence in HD patients with TCC. The OR values of the variables in the multivariate logistic regression were rounded to the assigned scores of the risk prediction model. The corresponding scores of each factor were summed in the training set to obtain the risk score. The receiver operating characteristic (ROC) curve was plotted, with area under the curve ( AUC) of 0.761(0.683-0.839) and maximum Youden index of 0.461, at which time the corresponding cut-off value was 6, with sensitivity of 90.0% and specificity of 56.1%. The model was validated in the validation set with AUC of 0.794(0.674-0.914) and cut-off value of 6, with sensitivity of 61.6% and specificity of 82.5%. Conclusions:Combined diabetes, history of catheter-related infection within 3 months, more than 4 times nursing interventions within 1 month, and central venous disease are the independent risk factors for CRBSI, and the prediction model based on the above factors has good efficacy in predicting the risk of CRBSI and can provide guidance for the prevention and treatment of CRBSI in HD patients.

Objective To study the relationships of TGFβ1 (-509C/ T, +869T/ C) and TGFβR2 (-875 G/ A) single nucleotide polymorphisms (SNPs) with colorectal cancer (CRC) in Chinese Han popu-lation in Shandong. Methods TGFβ1 -509C/ T and +869T/ C SNPs in a total of 490 patients with CRC were detected using gene chip. TGFβR2 -875 SNPs was analyzed using PCR-RFLP. TGFβ1 concentrations in serum samples were measured by ELISA. Immunohistochemistry was used to detect the expression of TGFβR2. The relationships of TGFβ1 (-509C/ T, +869T/ C) and TGFβR2 (-875 G/ A) SNPs with CRC were analyzed through a case-control study. Chi-square test or t test was used for statistical analysis. Rela-tive risk was estimated by odds ratio (OR) and 95％ confidence interval (95％ CI). Results No signifi-cant difference in genotype or allele frequency at TGFβ1 -509 / +869 was found between patients with CRC and healthy subjects (P>0. 05). The frequencies of TGFβR2 -875GG genotype and -875G allele in pa-tients with CRC were significantly higher than those in healthy subjects (-875GG: χ2 = 4. 65, P = 0. 031, OR=1. 32, 95％ CI=1. 03-1. 71; -875G: χ2 =4. 95, P=0. 026, OR=1. 29, 95％ CI=1. 03-1. 61). Com-pare with the healthy control group, higher frequencies of TGFβR2 -875GG genotype and -875G allele were also detected in rectal cancer ( -875GG: P = 0. 04, OR = 1. 39, 95％ CI = 1. 02-1. 95 and -875G: P =0. 045, OR=1. 32, 95％ CI = 1. 01-1. 73), tubular adenocarcinoma ( -875GG: P = 0. 004, OR = 1. 51, 95％ CI=1. 14-2. 00 and -875G: P=0. 003, OR=1. 45, 95％ CI=1. 14-1. 85) and highly differentiated tu-bular adenocarcinoma (-875GG: P=0. 003, OR=1. 68, 95％ CI=1. 19-2. 38 and -875G: P=0. 002, OR=1. 62, 95％ CI=1. 18-2. 21) groups. The serum TGFβ1 levels in TGFβR2 -875G carriers with CRC were significantly higher than those in TGFβR2 -875AA carriers in both CRC (t= -3. 42, P<0. 05) and healthy control (t= -5. 09, P<0. 001) groups. TGFβR2 expression in -875G carriers with rectal cancer was signifi-cantly lower than that in -875AA carriers with rectal cancer (P=0. 047) and healthy subjects (P=0. 027).Conclusion TGFβR2 -875GG might be a potential risk factor for CRC in Chinese Han population in Shandong and TGFβR2 - 875G might be a risk factor for rectal cancer and highly differentiated tubular adenocarcinoma.

Objective To study the correlations between genetic polymorphisms of TNF-α as well as IL-6 and susceptibility to colorectal cancer among Chinese Han people in Shandong province.Methods Single nucleotide polymorphisms (SNPs) of TNF-α-238G/A,-308G/A and IL-6-174G/C,-572G/C,-597G/A in 490 patients with colorectal cancer were analyzed by using gene chip.Concentrations of TNF-α and IL-6 in serum samples were measured by ELISA.A case-control study was conducted to analyze the correlations between SNPs of TNF-α-238G/A,-308G/A as well as IL-6-174G/C,-572G/C,-597G/A and susceptibility to colorectal cancer.Chi-square test or t test was used for statistical analysis.Relative risks were estimated based on the values of odds ratio (OR) and 95% confidence interval (95%CI).Results The frequency of TNF-α-308AA in patients with colorectal cancer was significantly higher than that in healthy subjects (x2 =6.15, P<0.05, OR=2.08, 95%CI=1.17-3.71), while the frequency of IL-6-572CC in patients with colorectal cancer was significantly lower than that in healthy subjects (x2 =4.97, P<0.05, OR=0.73, 95%CI=0.55-0.96).The frequency of TNF-α-308AA in patients with colon cancer (OR=2.31, 95%CI=1.17-4.55), tubular adenocarcinoma (OR=2.32, 95%CI=1.28-4.21), high (OR=2.05, 95%CI=1.01-4.15) or moderately differentiated adenocarcinoma (OR=5.88, 95%CI=1.79-19.30) was significantly higher than that in healthy subjects.The levels of serum TNF-α in TNF-α-308AA carriers with colorectal cancer were significantly higher than those in TNF-α-308G carriers with colorectal cancer (t=2.13, P<0.05) as well as those in healthy TNF-α-308AA carriers (t=2.13, P<0.05).The levels of serum IL-6 in colorectal cancer group were significantly higher than those in control group (t=6.74, P<0.001).Conclusion The SNPs of TNF-α-308 and IL-6-572 are associated with the occurrence and development of colorectal cancer in Chinese Han people in Shandong province.

OBJECTIVE:To evaluate the association between UGT1A1 gene polymorphism and irinotecan-induced 3-4 degree neutropenia,and to provide evidenced-based reference for clinical treatment. METHODS:Retrieved from CJFD,Wanfang data-base,VIP,PubMed,EMBase,Science direct and Cochrane library,related studies about UGT1A1*28 and UGT1A1*6 gene polymorphism and irinotecan-induced 3-4 degree neutropenia were collected. After data extraction and quality evaluation of included studies,Meta-analysis was conducted by using Review Man 5.3 software. RESULTS:A total of 29 studies were included,involv-ing 2408 patients. UGT1A1*28 includ wild genotype TA 6/6(UGT1A1*1/*1)and mutations genotype TA 6/7(UGT1A1*1/*28)、TA 7/7(UGT1A1*28/*28),UGT1A1*6 includ wild genotype GG and mutations genotype GA、AA. Results of Meta-analysis showed:the incidence of 3-4 degree neutropenia in UGT1A1*28 and UGT1A1*6 mutations genotype were significantly higher than wild genotype,with statistical significance [UGT1A1*28:OR=1.92,95%CI(1.52,2.44),P<0.001;UGT1A1*6:OR=2.49, 95%CI(1.46,4.26),P<0.001]. Using medium-dose and high-dose of irinotecan,the incidence of 3-4 degree neutropenia in UGT1A1*28 and UGT1A1*6 mutations genotype were significantly higher than wild genotype,with statistical significance [UGT1A1*28:OR=2.06,95%CI(1.57,2.70),P<0.001);UGT1A1*6:OR=1.92,95%CI(1.35,2.74),P<0.001]. Using low-dose of irinotecan,there was no statistical significance in the incidence of 3-4 degree neutropenia between UGT1A1*28,UGT1A1*6 mutations genotype and wild genotype [UGT1A1*28:OR=1.20,95%CI(0.70,2.08),P=0.51;UGT1A1*6:OR=3.19,95%CI (0.85,11.89),P=0.08]. CONCLUSIONS:Using medium-dose and high-dose of irinotecan,UGT1A1*28 and UGT1A1*6 muta-tions will increase the risk of severe neutropenia in cancer patients. Using low-dose of irinotecan,there is no clear correlation be-tween gene polymorphism and the neutropenia.

Objective To study the characteristics of IL-1B-31/-511 single nucleotide polymor-phisms (SNPs) and the variable number tandem repeat (VNTR) in intron 2 of the IL-1ra gene (IL-1RN) in patients with HCV-related liver diseases .Methods The concentration of IL-1βand IL-1ra in serum sam-ples was measured by ELISA assay .The SNPs of IL-1B gene (-31C/T,-511C/T) from 310 cases with HCV infection and 324 unrelated healthy controls were determined by using gene chip analysis , and the results for some randomly selected specimens were compared with those by using polymerase chain reaction -restriction fragment length polymorphism ( PCR-RFLP) assay.The VNTR polymorphism of IL-1RN intron 2 was ana-lyzed by PCR-RFLP assay.The serum level of alanine aminotransferase (ALT), an indicator of hepatocellu-lar injury, was detected by ROCHE cobas 8000 analyzer.HCV replication was measured by using specific fluorescence PCR .The genotypes of HCV were determined by direct nucleotide sequencing test .Results Compared with control group, the serum level of both IL-1β[(22.6 ±7.3) vs (13.7 ±4.2)] pg/ml and IL-1ra [(286.30 ±55.10) vs (185.55 ±48.32)] pg/ml were significantly increased in patients with HCV infection ( P<0.01 ) .There were two predominant genotypes identified among 310 patients including HCV 1b (75.5%) and HCV 2a (22.3%).The serum level of IL-1βand IL-1ra in IL-1B-511T carriers from four groups including case group , mild and moderate Hepatitis C group , severe Hepatitis C , and cirrhosis and hepatocellular carcinoma (HCC) group were significantly higher than those from IL-1B-511CC carriers and control group (P<0.05).The ratio of IL-1ra to IL-1βin all IL-1B-511T carriers with HCV infection were lower than those from healthy controls (P<0.05).IL-1B-511T carriers with HCV genotype 1b infec-tion showed a higher serum level of IL-1βas compared with those with HCV genotype 1a infection ( P<0.05).Compared with control group, they also showed an increase in IL-1ra level (P<0.05).There was no significant difference in the serum level of IL-1βamong IL-1B-511CC carriers from each group ( P>0.05).The frequency of IL-1B-511TT genotype (P<0.05, OR=1.55, 95% CI =1.10-2.18) and IL-1B-511T allele (P<0.05,OR=1.31,95% CI=1.05-1.63) in patients with HCV infection were signifi-cantly higher than those in healthy controls .IL-1B-511C/T SNP showed a significant association with the outcomes of HCV infection (P<0.005).Compared with IL-1B-511CC and IL-1B-511CT, IL-1B-511TT was a major risk factor for mild and moderate Hepatitis C [ OR=2.17 ( 1.48-3.19 ) ] , severe Hepatitis C [OR=2.11(1.05-4.26)], cirrhosis [OR=2.98(1.77-4.99)] and HCC [4.33(2.16-8.67)].IL-1B-511 T allele was significantly associated with mild and moderate Hepatitis C [ 1.80 ( 1.38-2.36 ) ] , severe Hepatitis C [1.80(1.08-3.01)], cirrhosis [2.62(1.76-3.89)] and HCC [3.49(1.96-6.23)].The fre-quency of IL-1B-511T allele showed significant difference among each group (P<0.005).No association was found between any of the other polymorphisms and HCV infection .Conclusion The serum level of IL-1βand IL-1ra were significantly associated with HCV infection .IL-1B-511T allele in patients with HCV in-fection up-regulated the serum level of IL-1β.IL-1B-511TT and IL-1B-511T allele were major risk factors for mild and moderate Hepatitis C, severe Hepatitis C, cirrhosis and HCC, but IL-1B-511CC/C had oppo-site effects.

This study was aimed to establish a high performance liquid chromatography (HPLC) method coupled with Evaporative Light-scattering Detector (ELSD) in order to develop the determination of fingerprint of terpene lactones in Ginkgo biloba tablets. An Agilent Extend-C18 (4.6 mm í 250 mm, 5 μm) was employed as the analysis column and the normal propyl alcohol-tetrahydrofuran-water (1:15:84) as mobile phase. The column temperature was 30℃. And the flow rate was 1.0 mL·min-1. HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS) was used to identify the common peaks. The fingerprint was further evaluated by chemometrics methods including principal component analysis (PCA), similarity analysis (SA) and hierarchical clus-tering analysis (HCA). The results showed that the precision, stability and repeatability of this method were favorable. Five common peaks were identified by LC/Q-TOF MS as ginkgolide J ( M ) , C , A , B and bilobalide , respectively . Fourteen batches of Ginkgo biloba tablets were determined. With the aid of PCA, SA and HCA, the common pattern of the fingerprint of terpene lactones was established. Samples were divided into 4 clusters by their quality differ-ence. It was concluded that the method established in this paper can be used for quality evaluation of terpene lac-tones in G ink go b ilob a tablets.

OBJECTIVETo develop a GC method for simultaneous determination of 4 compounds (atractylone, hinesol, beta-eudesmol and atractylodin) in Atractylodes lancea.

METHODA HP-1 capillary column (0.25 mm x 30 m, 0.25 microm) was used. The detector was FID:Inlet temperature was 250 degrees C. The detector temperature was 250 degrees C. The column temperature was set at 145 degrees C and held for 25 min after injection, then programmed at 10 degrees C x min(-1) to 250 degrees C and held for 10 min at the temperature. The carrying gas was nitrogen, split ratio was 40:1. Injection volume was 2 microL, Cluster analysis was performed by SPSS13.0 software.

RESULTThe linear ranges for atractylone, hinesol, beta-eudesmol and atractylodin were 0.0122. 32 (r = .9998), 0.008-1.68 (r = 0.9998), 0.009-1.76 (r = 0.9999), 0.016-3.20 g x L(-1) (r = 0.9997), respectively. The average recoveries (n = 3) of atractylone, hinesol, beta-eudesmol and atractylodin were 98.0%-99.0%, 97.7%-99.4%, 98.4%-99.2%, 97.8%-99.7%, respectively. The samples analyzed were divided into two classes.

CONCLUSIONThis method is simple, specific, repeatable and stable. It can be applied for the simultaneous determination of 4 compounds (atractylone, hinesol, beta-eudesmol and atractylodin) in A. lancea, which will provide the basis for the quality control of A. lancea. The contents of 4 active compounds were significantly different between geo-authentic and non-authentic producing areas.

Atractylodes ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; Furans ; analysis ; Plant Extracts ; chemistry ; Plant Oils ; analysis ; Quality Control ; Sesquiterpenes ; analysis ; Sesquiterpenes, Eudesmane ; analysis ; Spiro Compounds ; analysis