Objective To identify primary gout biomarkers. Methods Isobaric tags for relative and absolute quantitation (iTRAQ) technique combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to screen differentially expressed proteins, and to identify potential biomarkers by analysis of the biological process, cellular components, molecular functions, KEGG pathways and protein-protein interactions. Difference between two groups were measured byt test. Results We identified 95 differentially expressed proteins (50 up-regulated proteins and 45 down-regulated proteins, respectively), and 20 significant KEGG pathways. Among them, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), α-enolase (ENOA), phosphoglycerate kinase (PGK1), glucose-6-phosphate isomerase (G6PI) and moesin might play a role in the pathogenesis of primary gout. Conclusion iTRAQ technology can detect differentially expressed proteins from proteome, provides a strong theoretical basis for the study of biomarkers and evidence for the mechanisms in primary gout. However, further studies are needed.

The theory and application of small animal radiotherapy models is critical for the research and development of radiobiology and clinical radiotherapy.Considering universality and cost effects,mouse models are widely used to explore the radiobiological mechanisms of cancerous and normal tissues.In recent years,there has been tremendous progress in image-guided stereotactic radiotherapy equipment for small animals,which could simulate the human radiotherapy process.This article introduces stereotactic radiotherapy systems for small animals guided by different imaging modalities,such as cone-beam computed tomography and magnetic resonance imaging,and then reviews small animal fluorescence imaging technology and summarizes the application of different bioluminescence and fluorescence imaging equipment in small animal imaging systems.Finally,we put forward the prospect of optimization direction of radiotherapy equipment for small animals in future.

Objective To investigate the expression of microRNA-152 in hepatocellular carcinoma, and analyze the correlation of miR-152 expression with the clinicopathological characteristics of hepatocellular carcinoma(HCC) patients. Methods Real-time PCR was used to detect the expression of miR-152 in tissue specimens from HCC and the adjacent non-cancerous hepatic tissues collected from 56 patients who underwent surgical treatment for primary HCC. The association between the expression of miR-152 and the clinical pathological characteristics and prognosis of HCC patients was analyzed. Results The relative expression of miR-152 in hepatocellular carcinoma and corresponding adjacent non-carcinoma tissues were 0.616±0.041 and 0.768±0.042 (P=0.011). The expression of miR-152 was significantly associated with TNM stages (P=0.042), tumor differentiation (P=0.035), metastasis (P=0.048) and venous invasion (P=0.042). There was no significant association between the miR-152 expression and gender, age, AFP level, tumor number or tumor grade. The OS (P=0.035) and PFS (P=0.012) in the low miR-152 expression group were obviously shorter than those in the high miR-152 expression group. Multivariate Cox regression analysis showed that the low expression of miR-152, tumor recurrence and pathological classification were significant independent prognostic factors for the prognosis of hepatocellular carcinoma patients. Conclusion The expression of miR-152 is decreased in hepatocellular carcinoma tissues, and the low expression of miR-152 is significantly associated with TNM stages, tumor differentiation, metastasis, venous invasion and the prognosis of HCC patients.

OBJECTIVETo observe the inhibitory effect of 4-chlorobenzoyl berbamine (BBD9) on imatinib-resistant cell line K562 (K562/IR) in vitro and in vivo and explore the mechanisms.

METHODSThe IC50 of BBD9 and berbamine (BBM) was determined by MTT assay. The expressions of p210(Bcr-Abl), IKKa, cytoplasmic and nuclear NF-κBp65 were determined using Western blotting in K562/IR cells following a 48-h exposure to 0.5 µg/ml BBD9 or 8 µg/ml BBM. Flow cytometry was used to analyze the cell viability, apoptosis and necrosis; Western blotting was employed to determine the expressions of PARP, caspase-3, caspase-9 and LC3II in K562/IR cells exposed to different concentrations of BBD9 for 48 h. In nude mouse models bearing K562/IR cell xenograft, the tumor weight, tumor regression, and body weight changes of the mice were measured after treatments with 15 mg/kg and 30 mg/kg BBD9 and 100 mg/kg imatinib.

RESULTSThe IC50 of BBD9 and BBM was 0.73 µg/ml and 5.43 µg/ml, respectively. In K562/IR cell cultures, the expressions of p210(Bcr-Abl), IKKa and nuclear NF-κB p65 were all decreased following BBD9 and BBM treatments, but BBD9 produced more potent effect; cytoplasmic NF-κB p65 showed no obvious changes after the treatments. The cell apoptosis and necrosis increased with the concentrations of BBD9, which also dose-dependently increased the levels of cleaved caspase-3, csapase-9, PARP, and LC3II expression. In the tumor-bearing mouse model, BBD9 showed stronger effects than imatinib in reducing the tumor weight, promoting tumor regression, and increasing the body weight.

CONCLUSIONBBD9 can effectively inhibit the growth of K562/IR cells in vitro and in vivo by activating cell apoptosis, necrosis and autophage pathways, down-regulating expressions of p210(Bcr-Abl) and IKKa and suppressing the cytoplasm-to- nucleus translocation of NF-κBp65.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Benzamides ; Benzylisoquinolines ; pharmacology ; therapeutic use ; Drug Resistance, Neoplasm ; Female ; Fusion Proteins, bcr-abl ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; I-kappa B Kinase ; metabolism ; Imatinib Mesylate ; K562 Cells ; Liver Neoplasms, Experimental ; drug therapy ; metabolism ; Mice ; Mice, Nude ; Piperazines ; pharmacology ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; pharmacology ; Transcription Factor RelA ; metabolism ; Xenograft Model Antitumor Assays

Haizao Yuhu decoction (HYD) is a formula that has been used for approximately 500 years and famous for its efficiency in treating thyroid-related diseases in traditional Chinese medicine (TCM). HYD was first presented by Chen Shi-gong in a famous surgical monograph named Waike Zhengzong during the Ming Dynasty. We conducted the research to investigate the possible pharmacokinetic profile of different prescriptions of HYD in rats, in order to reveal the interactions of Haizao and Gancao drug pair with other herbs in HYD. Liquiritin, naringin, besperidin, peimine, peiminine liquiritigenin, glycyrrhizic acid, hergapten, nobiletin, osthole, glycyrrhetinic acid in blood samples were determined by UPLC-MS/MS. The result revealed tbat Haizao could enhance the peak concentration of glycyrrhizic acid. The other herbs in HYD may promote'the absorption of flavonoids in Gancao in normal rats, but inhibit the absorption of saponins and accelerate their metabolism. Gancao and Haizao drug pair could enhance the bioavailability of hesperidin, peimine, bergapten, nobiletin and osthole and prolong the elimination of peimine and naringin.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Magnetic Resonance Spectroscopy ; Male ; Mass Spectrometry ; Plasma ; chemistry ; Rats ; Rats, Wistar

Objective To investigated the clinical distributions and antimicrobial susceptibility of Streptococcus agalactia strains isolated from the patients .Methods The identification and susceptibility of the strains were mainly measured by automatic VITEK‐Ⅱ system ,the K‐B disc diffusion tests were used for the resistance test of erythromycin ,meropenem ,and D‐test .Results The iso‐lates were mainly from urine (63 .1% ) ,genital tract(7 .8% ) and wound secretion(6 .7% ) .They were obtained from patients in dif‐ferent situations ,including 110 patients who were older than 50 years old (61 .5% ) ,113 female patients (63 .1% ) ,12 gravidas (6 .7% ) ,3 vertical transmitted newborns(1 .7% ) ,and 82 patients with cancer ,undergoing chemo radiotherapy ,with diabetes ,tuber‐culosis or after operations(45 .8% ) .The resistant rates of the isolated Streptococcus agalactia to erythromycin and clindamycin were 42 .9% -93 .3% and 41 .9% -80 .0% respectively .The positive rate of D‐test was 4 .1% .The strains were highly resistant to tet‐racycline(>80% ) ,while the resistance to penicillin was below 10% except in 2008 .All isolates were susceptible to vancomycin and meropenem .Only one strain was resistant to Quinupristin‐dalfopristin .Conclusion Streptococcus agalactia infection in adults most‐ly cause genitourinary tract ,skin and soft tissue infections .There were more females than males with Streptococcus agalactia infec‐tion .Penicillin andβ‐lactams are still the first choice for the treatment .Erythromycin ,clindamycin and tetracycline should be used with caution under the guidance of laboratory susceptibility test results .

To investigate changes of 14-3-3beta from apoptosis induced by PrP106-126 polypeptide, HeLa cell was incubated with PrP106-126 for 4h or 8h. Nucleus changes and the expression of PARP were detected differently by Hoechst staining and Western blotting. Expressing of protein and mRNA from 14-3-3beta was determined by Western blotting and Real-time PCR. The results show that typical nucleus pyknosis and chip of apoptosis and degradation of PARP were induced by PrP106-126 peptide in HeLa cells. Degradation of 14-3-3beta appeared in apoptosis groups induced by PrP106-126 peptide. However, 14-3-3beta mRNA did not display any changes in apoptosis groups. This study indicated that degradation of antiapoptosis protein 143-3beta induced by PrP106-126 peptide may be one of pathogenesis mechanism of prion disease.
14-3-3 Proteins ; metabolism ; Apoptosis ; drug effects ; HeLa Cells ; Humans ; Peptide Fragments ; pharmacology ; Prions ; pharmacology ; Proteolysis ; drug effects

Objective:To investigate the significance of serum fibroblast growth factor 19 (FGF19), Klotho and fibroblast growth factor receptor 4 (FGFR4) protein expression in patients with primary biliary cirrhosis (PBC).Methods:Sixty-three patients with PBC who received treatment in Ningbo Huamei Hospital, University of Chinese Academy of Sciences between August 2017 and July 2020 were included in the PBC group. An additional 51 healthy patients who concurrently received physical examination in the same hospital were included in the control group. Serum FGF19, Klotho and FGFR4 protein expression were determined by enzyme linked immunosorbent assay.Results:Serum FGF19 and FGFR4 protein in PBC group were (178.86 ± 21.28) ng/L and (2.96 ± 0.47) ng/L, respectively, which were significantly higher than those in the control group [(69.93 ± 12.12) ng/L, (1.21 ± 0.35) ng/L, t = 32.51, 27.98, both P < 0.05]. Klotho protein expression in the PBC group was significantly lower than that in the control group [(3.25 ± 0.89) μg/L vs. (9.67 ± 1.53) μg/L, t = 22.08, P < 0.05]. Serum levels of alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase in the PBC group were (84.25 ± 13.24) U/L, (71.82 ± 10.35) U/L, (278.93 ± 32.45) U/L, respectively, which were significantly higher than those in the control group [(23.76 ± 3.42) U/L, (23.10 ± 4.53) U/L, (76.81 ± 16.36) U/L, t = 31.75, 31.26, 40.48, all P < 0.05]. The receiver operating characteristic curve analysis showed that the sensitivity and specificity of FGF19 in the diagnosis of PBC were 76.67% and 61.90%, respectively, they were 58.82% and 66.67% for Klotho protein diagnosis, 54.55% and 76.67% for FGFR4 protein. Pearson analysis revealed that there was a positive linear relationship between FGF19 and FGFR4 protein ( r = 0.78, P < 0.05), while there was a negative linear relationship between Klotho protein and FGFR4 protein ( r = -0.72, P < 0.05). Conclusion:In patients with PBC, serum FGF19 and FGFR4 protein levels are increased, while Klotho protein level is decreased. There is a positive linear relationship between FGF19 and FGFR4 protein, and there is a negative linear relationship between Klotho protein and FGFR4 protein. This study is highly innovative and scientific.

OBJECTIVETo explore the role of reversal multidrug resistance (MDR) using short hairpin RNA (shRNA) expression vectors in multidrug resistance human leukemia cell line K562/ADM.

METHODSThe oligonucleotides with 19-mer hairpin structure were synthesized. The shRNA expression vectors were constructed and introduced into K562/ADM cells. Expression of mdr1 mRNA was assessed by RT-PCR, and P-gp expression was determined by Western blot. The apoptosis and sensitivity of the K562/ADM cells to doxorubicin were quantified by flow cytometry and methyl thiazolyl tetrazolium (MTT) assays, respectively. Cellular daunorubicin accumulation was assayed by laser confocal scanning microscope (LCSM).

RESULTSIn positive clones of K562/ADM cells stably transfected with pSilencer 3.1-HI neo mdr1-A and mdr1-B shRNA expression vectors, RT-PCR showed that mdr1 mRNA expression was significantly reduced to 35.9% (P < 0.05), 27.5% (P < 0.01), respectively. Western blot showed that P-gp expression was significantly and specifically inhibited. Resistance against doxorubicin was decreased from 79-fold to 38-fold (P < 0.05), 30-fold (P < 0.01) respectively. Furthermore, the fluorescence intensity of K562/ADM cells was increased significantly compared with the control. shRNA vectors significantly enhanced the cellular daunorubicin accumulation. The percent of the apoptosis cell was significantly enhanced to 18.1% (P < 0.05) , 54.4% (P < 0.01) respectively.

CONCLUSIONSshRNA expression vectors can effectively reverse MDR, and restore the sensitivity of drug-resistance K562/ADM cells to conventional chemotherapeutic agents.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Apoptosis ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Genetic Vectors ; Humans ; K562 Cells ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo evaluate the prognostic significance of a new grading and scoring system (based on the new IASLC/ATS/ERS classification) in stage I pulmonary adenocarcinoma, as compared with the WHO grading system.

METHODSThe clinicopathologic characteristics of 125 patients with stage I pulmonary adenocarcinoma primarily treated by surgical resection were reviewed retrospectively. All cases were classified according to the new IASLC/ATS/ERS classification and graded into three prognostic groups based on the new classification, the Sica scoring system and the WHO grading system, respectively. The differences in prognosis of the three groups were analyzed.

RESULTSThere was a statistically significant correlation between the new grading system and the WHO grading system (P = 0.000). Both of them showed negative correlation with overall survival. The new scoring system however better correlated with disease recurrence and/or metastasis (P = 0.855, P = 0.073 versus P = 0.011). According to univariate Log-rank test, the prognosis correlated with tumor size (P = 0.004), clinical stage (P = 0.000), the WHO grading (P = 0.020), the new grading system (P = 0.000), the new scoring system (P = 0.000), vascular invasion (P = 0.021), and recurrence and/or metastasis (P = 0.000). The Cox regression analysis demonstrated that clinical stage (P = 0.014), the new grading system (P = 0.047), the new scoring system (P = 0.043), and recurrence and/or metastasis (P = 0.018) were significantly independent poor prognostic factors.

CONCLUSIONSThe new grading and scoring system shows good correlation with the WHO grading system. Compared with the WHO grading system, the new scoring system based on the new IASLC/ATS/ERS classification provides valuable information in categorizing stage I pulmonary adenocarcinoma cases with different risks of disease recurrence, tumor metastasis and prognosis.

Adenocarcinoma ; classification ; pathology ; surgery ; Adenocarcinoma, Mucinous ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoma in Situ ; pathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; classification ; pathology ; surgery ; Male ; Middle Aged ; Neoplasm Grading ; methods ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Research Design ; Retrospective Studies ; Societies, Medical ; Young Adult