Objective: The aim of this study is to evaluate the commutability of 16 processed materials for 17-hydroxyprogesterone by using 2 commutability assessment approaches. Methods: 52 serum specimens were collected in Clinical Laboratory Department of Beijing Hospital from February 2018 to June 2019. According to the report of the Clinical and Laboratory Standards Institute (EP14-A3) document and the recommendations of the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) working group on commutabilityassessment, serum 17-hydroxyprogesterone isotope diluent chromatogram tandem mass spectrometry (ID-LC/MS/MS) was used for comparison. Three clinical routine analysis systems (1 radioimmunoassay, 2 LC/MS analysis methods) were used to determine the concentration of 17-hydroxyprogesterone in 52 human serum samples and 16 processed materialsfor commutabilityassessment. Results: Combined with the results of the two commutability assessment, all accuracy verification materials and national steroid hormone standards showed good commutability in the LC/MS analysis system, and 6/9 EQA materials showed commutability in the three routine analysis systems.All materials showed good commutability in the LC/MS analysis system of bias difference method. Conclusions The two kinds of commutability assessment results are different. Bias difference method has more clinical value, but it has certain application limitations. The use of fresh frozen human serum as a quality assessment materialfor serum 17-hydroxyprogesterone is meets the commutability requirement.

Objective To explore the role and mechanism of inducible nitric oxide synthase inhibitor 1 400W in alleviating ischemia-reperfusion injury of human intrahepatic bile duct epithelial cells.Methods Human intrahepatic bile duct epithelial cells (HIBEC) in logarithmic phase were inoculated into culture plate at an appropriate density.The samples were randomly divided into control group (group C),ischemiareperfusion group (group I/R) and ischemia-reperfusion + 1 400W group (group I/R + 1 400W).Group C was cultured routinely;cells in I/R and I/R + 1 400W groups were placed in a three-gas incubator for 12h for simulating ischemia and then normal culture for 6h for simulating reperfusion.The I/R + 1 400W group had a final concentration of 100 μmol/L of 1 400W before ischemia and hypoxia.After reperfusion,cells and culture medium were collected,CCK 8 was used for detecting cell vitality,microplate method for detecting the content of lactate dehydrogenase (LDH) in culture medium,AnnexinV-FITC/PI double stain for detecting apoptosis level,Western blot for analyzing the expressions of endoplasmic reticulum stress (ERS)related protein cysteinyl aspartic acid protease 12 (caspase-12),glucose regulatory protein 78 (GRP78) C/EBP homologous protein (CHOP) and inducible nitric oxide synthase (iNOS).Results As compared with group C,cell viability significantly decreased in I/R and I/R+ 1 400W groups (53.8％ ± 2.3％ vs.100％,66.5 ％ ± 2.8 ％ vs.100 ％) (P＜0.05) while LDH increased markedly in cell culture medium (287.4 ±9.0U/L vs 120.2 ± 8.7U/L,212.0 ± 8.3U/L vs 120.2 ± 8.7U/L) (P＜0.05).Apoptosis accelerated markedly (41.5％±2.3％ vs5.2％±0.5％,32.7％± 1.8％ vs 5.2％±0.5％) (P＜0.05) and the expressions of caspase-12,GRP78,CHOP and iNOS spiked (P＜0.05);as compared with I/R group,cell viability of I/R+ 1 400W group rose while LDH,apoptosis level,caspase-12,GRP78 and CHOP declined in cell culture medium (P＜0.05).Conclusions 1 400W may alleviate ischemia-reperfusion injury of human intrahepatic bile duct epithelial cells and its mechanism may be correlated with a suppression of endoplasrnic reticulum stress.

Objective To investigate the effects of curcumin on the growth and apoptosis of human hepatoma HCC-LM3 cells and explore the molecular mechanisms.Methods The human hepatoma HCC-LM3 cells were treated with different concentrations of curcumin and the cell growth inhibition rate was detected by CCK-8.The effect of curcumin on human hepatoma HCC-LM3 cells was observed.Refer to the relevant literature,the human hepatoma HCC-LM3 cells were treated with the concentration of 2.5,5.0,10.0,15.0,20.0,40.0,60.0 μmol/L of curcumin for 48 hours,taking the 0 μmol/L curcumin as control group,and the cell growth inhibition rate was detected by CCK-8.According to the results of CCK-8,selecting the concentration of 0 μmol/L as control group and the concentration of 10.0,20.0,40.0 μmol/L as experimental groups,which has significant difference on growth inhibition rates.Cell cloning assay was used to detect cell cloning ability,Flow cytometry was used to detect apoptosis,and Western blotting to detect the protein expression levels of Mcl-1,Bax,Bcl-2 and Bcl-xL.The measurement data were expressed in ((x) ± s),and the single factor analysis of variance was used for comparison between groups.Results CCK-8 assay showed that with treated by the concentration of 2.5,5.0,10.0,15.0,20.0,40.0,60.0 μmol/L,the growth inhibition rates were(6.71 ± 3.45)％,(12.33 ± 5.02)％,(20.07 ± 5.60)％,(57.80 ±7.34)％,(78.37 ±6.53)％,(91.73 ±6.14)％ and (96.18 ±3.45)％,suggesting that curcumin could inhibit the growth of human hepatoma HCC-LM3 cells in a dose-dependent manner.Cell clone formation experiment showed that curcumin could inhibit the clone of the human hepatoma HCC-LM3 cells,and the clone of the cells was inhibited significantly when the concentration of the curcumin was over 20.0μmol/L.The result of Annexin V-FITC/PI double staining analysis showed that the apoptotic rates of experimental groups and control groups were (5.20 ± 1.44) ％,(9.90 ± 3.31) ％,(55.67 ± 5.29) ％,(79.63 ±4.71)％,with all the apoptotic rates of experimental group over the control groups (P ＜0.05),suggesting curcumin could induce the apoptosis of human hepatoma HCC-LM3 cells.The Westen blotting showed that curcumin increased the expression of Bax protein while decreasing expression of Mcl-1 protein significantly in concentration-dependent manner (P ＜ 0.05),but have no effect on the expression of Bcl-2 and Bcl-xL proteins.Conclusion Curcumin could inhibit the proliferation and clone of human hepatoma HCC-LM3 cells,and induce apoptosis in a dose dependent manner.

Objective To investigate the complications of High Intensity Focused Ultrasound (HIFU) in the treatment of primary hepatic carcinoma,looked forward to improving the safety of the treatment of HIFU.Methods Retrospectively analyzed the clinical data of 165 patients with primary liver cancer treated by High Intensity Focused Ultrasound in the First Affiliated Hospital of Zhengzhou University from September 2014 to December 2009,to summarise the complications of this therapeutic method and analysis the related treatment measures.Results HIFU treatment of primary liver cancer common complications included the treatment of skin soft tissue injury (118/165),a transient injury of liver function (83/165),postoperative fever (81/165),self-limiting pleural effusion (42/165) and postoperative pain (33/165);The severe complications included rib fractures(2 cases),organ perforation injury (1 case),Cancer rupture hemorrhage (1 case)and Skin 1Ⅱ(5 cases),Ⅲ(1 case) degree burn;Serious consequences were not caused by the complications which were actively treated.Conclusion The treatment of primary liver cancer by High Intensity Focused Ultrasound has less serious complications and high safety.

Pentraxin 3 (PTX3) was identified as a marker of the inflammatory response and overexpressed in various tissues and cells related to cardiovascular disease. Honokiol, an active component isolated from the Chinese medicinal herb Magnolia officinalis, was shown to have a variety of pharmacological activities. In the present study, we aimed to investigate the effects of honokiol on palmitic acid (PA)-induced dysfunction of human umbilical vein endothelial cells (HUVECs) and to elucidate potential regulatory mechanisms in this atherosclerotic cell model. Our results showed that PA significantly accelerated the expression of PTX3 in HUVECs through the IkappaB kinase (IKK)/IkappaB/nuclear factor-kappaB (NF-kappaB) pathway, reduced cell viability, induced cell apoptosis and triggered the inflammatory response. Knockdown of PTX3 supported cell growth and prevented apoptosis by blocking PA-inducted nitric oxide (NO) overproduction. Honokiol significantly suppressed the overexpression of PTX3 in PA-inducted HUVECs by inhibiting IkappaB phosphorylation and the expression of two NF-kappaB subunits (p50 and p65) in the IKK/IkappaB/NF-kappaB signaling pathway. Furthermore, honokiol reduced endothelial cell injury and apoptosis by regulating the expression of inducible NO synthase and endothelial NO synthase, as well as the generation of NO. Honokiol showed an anti-inflammatory effect in PA-inducted HUVECs by significantly inhibiting the generation of interleukin-6 (IL-6), IL-8 and monocyte chemoattractant protein-1. In summary, honokiol repaired endothelial dysfunction by suppressing PTX3 overexpression in an atherosclerotic cell model. PTX3 may be a potential therapeutic target for atherosclerosis.
Apoptosis/drug effects ; Atherosclerosis/chemically induced/*drug therapy/immunology/pathology ; Biphenyl Compounds/chemistry/isolation & purification/*pharmacology ; C-Reactive Protein/*genetics/immunology ; Down-Regulation/drug effects ; Drugs, Chinese Herbal/chemistry/isolation & purification/*pharmacology ; Human Umbilical Vein Endothelial Cells ; Humans ; I-kappa B Kinase/*immunology ; Lignans/chemistry/isolation & purification/*pharmacology ; Magnolia/chemistry ; Palmitic Acid ; Protein-Serine-Threonine Kinases/*immunology ; Serum Amyloid P-Component/*genetics/immunology ; Signal Transduction/drug effects

Objective To study the efficacy and the safe dosage of high intensity focused ultrasound (HIFU) ablation in pancreatic cancer.Methods From November 2010 to May 2013,21 patients with advanced pancreatic cancer were treated by HIFU at the First Affiliated Hospital of Zhengzhou University.These patients who were randomly divided into two groups (10 and 11 patients respectively),were given a low-power (100～249 W) treatment and a high power (250 ～350 W) treatment.These two groups of 21 patients received a total of 25 times of HIFU treatment (3 patients received twice of low-power treatment,while 1 patient received twice of high-power treatment).The two groups were compared by analyzing the treatment parameters (average power,total treatment time,treatment total energy,treatment volume,etc.) and volume of tumour response as shown on postoperative imaging (CT or MRI) examinations.Also,the complications,degree of pain relief and survival were compared.The energy efficiency factor (EEF) and the ablation ratio were calculated.A preliminary study was conducted on the relationship of the ultrasound dose and the ablation effect of HIFU treatment for pancreatic cancer.Results (1) The EEF of the high-power group (≥250 W) and the low-power group (＜ 250 W) were (10.39 ± 5.71) J/mm3 and (21.62 ± 9.81) J/mm3,the former group was significantly lower than the latter group (P ＜0.05) ; the ablation ratio of the high-power group was higher than the low-power group,(91.52 ± 4.18)％ versus (51.59 ± 7.66)％ respectively,the difference was statistically significant (P ＜ 0.001).(2) The efficiency factor and the ablation volume for the HIFU treatment showed a linear trend,and both were negatively correlated (Pearson correlation coefficient r =-0.485,P ＜ 0.05).(3) There was no serious complication after the HIFU treatment.In the low-power group,six of ten patients were alleviated of his pain (60％) ; the CA19-9 decreased in four of ten patients after HIFU treatment (40％).In the high-power group,nine of eleven patients were significantly relieved of pain after treatment (82％),the CA19-9 decreased in five of nine patients after HIFU treatment (56％).(4) On Kaplan-Meier survival analysis,HIFU treatment of patients with pancreatic cancer,the median survival was 8 months and 9 months in the low-power group and high power group,respectively (Log-rank test x2 =0.05,P =0.944).Conclusion During HIFU treatment of patients with pancreatic cancer,if the ultrasound power was between 250 W and 350 W,there was a higher proportion of tumor ablation,but with no serious complications.Thus,this dose was safe.

Objective To compare the curative effect of 360° arthroscopic denervation and routine arthroscopic debridement in treatment of knee osteoarthritis.Methods A total of 85 patients were included in the study after excluding the 5 follow-up dropout among the 90 patients with knee osteoarthritis treated between May 2006 and May 2011.Subsequently,the odd and even quality of the mantissa of admission number was used to determine the groups:Group A,42 patients underwent routine arthroscopic debridement; Group B,43 patients underwent arthroscopic debridement plus 360 degree denervation.Traumatic and degenerative arthritis occurred in 30 and 55 patients respectively.There were 34 male and 51 female patients whose mean age was 63 years (range,52-73 years).Western Ontario and McMaster University Osteoarthritis Index (WOMAC),MOS item short from health survey (SF-36),and visual analogue score (VAS) were compared between the two groups before and after operation.Results WOMAC score and SF-36 score of the two groups improved after surgery.At postoperative 6 months,12 months,and 24 months,WOMAC in Group B scored 81.0,78.5,and 82.4 respectively,lower than 86.8,83.8,and 91.7 in Group A (P ＜ 0.05) ; SF-36 in group B scored 47.4,46.3,and 44.4 respectively,higher than 43.9,41.4,and 39.5 in group A (P ＜ 0.05).At postoperative 1 week,VAS of group B was 1.6 points,lower than 2.6 points in group A (P ＜ 0.05).There was no statistical difference between the two groups in postoperative swelling of joint (P ＞ 0.05).Conclusions According to the area involved in knee osteoarthritis and characteristics of innervation of knee joint synovium,arthroscopic 360° denervation provides thorough debridement and removes as many innervation of knee joint synovium.Arthroscopic 360° denervation is superior to the routine arthroscopic debridement in improvement and duration of the symptoms.

Objective To investigate the effects of RNA interference targeting EphA7 gene on the growth of SMMC-7721 cell xenograft in nude mice.Methods Recombinant plasmid of EphA7 gene-targeting siRNA was transfected into hepatic cancer SMMC-7721 cells by LipofectamineTM2000,comparing with the empty vector transfected group,untransfected group and control group.The nude mice tumor model was established by subcutaneous injection of hepatic cancer cells in the left upper limb of the mice.Control group was injected with PBS as blank.Real-time PCR,immunohistochemistry and Western blot were employed to detect the mRNA and protein expressions of EphA7 in tumor tissues.The tumor formation time,tumor mass and weight of tumor were also considered in the analysis.Results About 9 ～ 12 days after the injection of tumor cells,the xenograft tumor formation can be observed around the injection site except the control group.35 days after tumor formation,there were obvious decreases in the tumor growth rate,tumor mass,as well as tumor weight in transfected group,comparing with empty vector transfected group and untransfected group (P ＜0.05).Transfection of RNA interference can inhibit the growth of xenograft tumor by 55％.Immunohistochemistry tests showed that there were less cells with positive staining of EPHA7 protein in transfected group,and the staining was lighter as pale yellow,in contrast with the untransfected group and the empty vector transfected group.Real-time PCR and Western blot revealed that the expression of EphA7 mRNA and EPHA7 protein of transfected group were significantly lower than those of untransfected group and empty vector trausfected group with statistically significance (P ＜ 0.05).Conclusion Silencing EphA7 gene with RNA interference can effectively inhibit the growth of SMMC-7721 cell in nude mice,which is expected to become a new target for gene therapy of hepatic cancer.

Objective To study the efficacy and safety of high intensity focused ultrasound (HIFU) treatment for advanced pancreatic cancer.Methods In this study,25 patients with advanced pancreatic cancer received high-intensity focused ultrasound (HIFU) treatment.Liver and kidney function,CA19-9 levels,tumor size changes,pain relief,survival rate before and after treatment were evaluated.Results The blood routine test,liver and kidney function,blood amylase did not alter significantly after HIFU treatment in all patients.The CA19-9 level of 12 patients decreased.The appetite of 15 patients improved,5 patients with body weight gain after HIFU treatment.Pain was relieved after HIFU treatment in 18 cases,pain relief rate was 72％ (18/25).In 15 cases tumor ablation volume ＞ 90％ after HIFU treatment,5 patients with tumor ablation volume ＞ 50％,tumor ablation effective rate was 80％ (20/25).There were no major complications such as acute pancreatitis,gastrointestinal injury after HIFU treatment.After HIFU treatment,the median survival period was 8 months,1 year survival rate was 30％.Conclusions High-intensity focused ultrasound is a safe and effective method of palliative treatment for advanced inoperable pancreatic cancer.

Cytotoxic T cells (CTLs) play a key role in the control of Hepatitis B virus (HBV) infection and viral clearance. However, most of identified CTL epitopes are derived from HBV of genotypes A and D, and few have been defined in virus of genotypes B and C which are more prevalent in Asia. As HBV core protein (HBc) is the most conservative and immunogenic component, in this study we used an overlapping 9-mer peptide pool covering HBc to screen and identify specific CTL epitopes. An unconventional HLA-A2-restricted epitope HBc141-149 was discovered and structurally characterized by crystallization analysis. The immunogenicity and anti-HBV activity were further determined in HBV and HLA-A2 transgenic mice. Finally, we show that mutations in HBc141-149 epitope are associated with viral parameters and disease progression in HBV infected patients. Our data therefore provide insights into the structure characteristics of this unconventional epitope binding to MHC-I molecules, as well as epitope specific CTL activity that orchestrate T cell response and immune evasion in HBV infected patients.
Adult ; Amino Acid Sequence ; Animals ; Binding Sites ; Epitopes ; chemistry ; immunology ; metabolism ; Female ; Genotype ; HEK293 Cells ; HLA-A2 Antigen ; metabolism ; Hepatitis B Core Antigens ; chemistry ; immunology ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Hydrogen Bonding ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Middle Aged ; Molecular Dynamics Simulation ; Mutation ; Protein Binding ; Protein Structure, Tertiary ; T-Lymphocytes, Cytotoxic ; immunology ; metabolism