Objective:To isolate Yersinia enterocolitica phage and study its biological and genomic characteristics, and explore its application value. Methods:Phages were isolated from cecum of Apodemus chevrieri in the plague focus of wild rodent in Lijiang employing Yersinia enterocolitica (CMCC52301) as the host strain. After purification, proliferation and concentration, the morphology was observed by transmission electron microscope, and the host spectrum of the phage was determined by double-layer plate method. Phage DNA was extracted and sequenced, and the raw sequences data were assembled and visually analyzed, also performed genome functional annotation, phage classification status, phylogenetic tree, genome collinearity analysis, etc. Results:One strain of Yersinia enterocolitica phage was isolated and named vB_Yen_YN301-151, which had a head and a contractile tail. The phage only lysed Yersinia enterocolitica, and could not lyse other tested strains. And at a lysis temperature of 37 ℃, more transparent plaques were observed compared to at a lysis temperature of 21 ℃. The gene structure of this phage was double-stranded DNA, with a genome length of 51 176 bp and a GC content of 46.96%, and 95 genes were predicted to encode 15 structural morphological related proteins, 9 proteins involved in replication and metabolism, 3 DNA assembly functional proteins, 1 phage lysis related protein, and 67 hypothetical proteins. It was identified that the phage belong to the Drexlerviridae family of the Caudoviricetes class, clustered with vB_YenM_534, vB_Yen_X1, vB_YenM_281, and vB_YenM_531 in the phylogenetic tree and exhibited good collinearity with PY100, vB_YenM_ 25, and vB_Yen_X1. Conclusions:The successfully isolated vB_Yen_YN301-151 is a novel phage belonging to the Drexlerviridae family, and its gene encoded proteins are mostly hypothetical proteins. It only lyses Yersinia enterocolitica, with host specificity and has the potential to be used as diagnostic phage for screening Yersinia enterocolitica.

Objective:To isolate Yersinia enterocolitica phage and study its biological and genomic characteristics, and explore its application value. Methods:Phages were isolated from cecum of Apodemus chevrieri in the plague focus of wild rodent in Lijiang employing Yersinia enterocolitica (CMCC52301) as the host strain. After purification, proliferation and concentration, the morphology was observed by transmission electron microscope, and the host spectrum of the phage was determined by double-layer plate method. Phage DNA was extracted and sequenced, and the raw sequences data were assembled and visually analyzed, also performed genome functional annotation, phage classification status, phylogenetic tree, genome collinearity analysis, etc. Results:One strain of Yersinia enterocolitica phage was isolated and named vB_Yen_YN301-151, which had a head and a contractile tail. The phage only lysed Yersinia enterocolitica, and could not lyse other tested strains. And at a lysis temperature of 37 ℃, more transparent plaques were observed compared to at a lysis temperature of 21 ℃. The gene structure of this phage was double-stranded DNA, with a genome length of 51 176 bp and a GC content of 46.96%, and 95 genes were predicted to encode 15 structural morphological related proteins, 9 proteins involved in replication and metabolism, 3 DNA assembly functional proteins, 1 phage lysis related protein, and 67 hypothetical proteins. It was identified that the phage belong to the Drexlerviridae family of the Caudoviricetes class, clustered with vB_YenM_534, vB_Yen_X1, vB_YenM_281, and vB_YenM_531 in the phylogenetic tree and exhibited good collinearity with PY100, vB_YenM_ 25, and vB_Yen_X1. Conclusions:The successfully isolated vB_Yen_YN301-151 is a novel phage belonging to the Drexlerviridae family, and its gene encoded proteins are mostly hypothetical proteins. It only lyses Yersinia enterocolitica, with host specificity and has the potential to be used as diagnostic phage for screening Yersinia enterocolitica.

Objective To investigate the anti-estrogenic activity of bisphenol A and its substitutes, and to analyze the relevant mechanisms. Methods Bisphenol A and its three most widely used substitutes (bisphenol S, bisphenol F and bisphenol AF) were selected as the docking ligand molecules, and estradiol was used as the control ligand molecule. The ligand molecules docking was simulated with estrogen receptor (ER) α and ERβ using AutoDock software. Results Bisphenol A forms a hydrogen bond with ERα at the His474 residue and with ERβ via three hydrogen bonds at Leu260, His428, and Asn431 residues. Similar to bisphenol A, bisphenol S, bisphenol F, bisphenol AF and estradiol primarily interact with ERα and ERβ through hydrophobic interactions and hydrogen bonds, but with varying optimal binding sites and affinities. The binding forces of the optimal binding sites for bisphenol A, bisphenol F, bisphenol AF, bisphenol S and estradiol with ERα were -4.15, -4.19, -2.73, -4.62 and -5.37 kcal/mol, respectively, and with ERβ were -3.76, -3.91, -2.86, -3.93, and -4.98 kcal/mol, respectively. The affinity ranking for two ERs with these five molecules from high to low was estradiol > bisphenol S> bisphenol F> bisphenol A > bisphenol AF. Conclusion The affinity between bisphenol compounds with ERα and ERβ is mainly based on the hydrophobic interaction with non-polar residues of the receptor and hydrogen bonding with key residues. Bisphenol S, bisphenol F and bisphenol AF showed similar or even stronger endocrine disrupting effects than bisphenol A.

Objective:To study the effectiveness and safety of great occipital nerve decompression in great occipital neurogenic intractable migraine.Methods:Fifty-one patients with great occipital neurogenic intractable migraine admitted to our hospital from December 2018 to December 2019 were chosen in our study. All patients were confirmed to be of great occipital neurogenic intractable migraine by blocked test and all patients underwent minimally invasive decompression of great occipital nerve. Preoperative and postoperative questionnaires were used to compare the severity of pain and frequency of pain episodes, and the prognoses of these patients were evaluated at the last follow-up.Results:Fifty-one patients were followed up for (12.02±3.36) months after surgery; at last follow-up, 10 patients were cured, 31 patients had obvious effect, 6 patients got improvement, 2 patients had poor effect, and 2 patients had no effect, with total effective rate of 80.40% (41/51). There were significant differences between before surgery and after surgery in the pain visual analog scale scores (7.47±0.76 vs.1.75±1.91) and frequency of pain episodes ([9.00±1.74] times/month vs. [2.82±2.47] times/month, P<0.05). Seven patients had temporary numbness at the surgical site after surgery, but 6 patients improved about 3 months after surgery and only one patient had persistent numbness. Conclusion:Great occipital nerve decompression is an effective and safe treatment method for great occipital neurogenic intractable migraine.

Objective:To investigate the expression of Macrophage migration-inhibitory factors (MIF) in hepatocellular carcinoma (HCC) tissues and its interaction with ERK1/2 signaling pathway, so as to establish a theoretical basis for further studying the molecular mechanism of MIF promoting HCC.Methods:From February 2020 to August 2021, 52 cases of hepatocellular carcinoma (HCC) tissues based on hepatitis B cirrhosis (HBV-LC) and 52 cases of adjacent tissues in Tianjin Medical University Cancer Hospital and 940th Hospital of Joint Logistic Support Force of PLA were collected as the experimental group, including 39 males and 13 females, aged 35-65 years. And 20 cases of normal liver tissue were selected as the control group. Immunohistochemistry was used to detect the expression of MIF, ERK1/2 and p-ERK1/2 proteins in liver tissues of the two groups, and in situ hybridization was used to detect the expression of ERK1/2 nucleic acid in liver tissues of the two groups.HepG2 HCC cells and L-02 normal hepatocytes were co-cultured with different concentrations of rMIF, the expression and phosphorylation levels of ERK1/2 and JNK1 proteins in the two kinds of liver cells were detected by Western-blot, and the expression levels of ERK1/2 nucleic acids in the two kinds of liver cells were detected by RT-PCR. One-way ANOVA was used for measurement data and χ 2 test was used for counting data. Results:The expressions of MIF, ERK1/2, p-ERK1/2 and ERK1/2 mRNA were significantly increased in HCC and para-cancer tissues (the expression of MIF in HCC group was 78.8%, and that in adjacent group was 75.0%; ERK1/2 80.8% in HCC group and ERK1/2 71.8% in paracancerous group. The expression of p-ERK1/2 75.0 % in HCC group and 46.2% in paracancerous group were respectively detected. ERK1/2 mRNA was expressed in HCC group 76.9%, ERK1/2 mRNA expression in paracancerous group 78.8%), and the differences were statistically significant compared with normal liver tissues ( P<0.05), but there was no significant difference between HCC and para-cancer tissues ( P>0.05). The expressions of ERK1/2, p-ERK1/2 and ERK1/2 mRNA in HepG2 HCC cells were significantly increased with the increase of rMIF concentration, and the increase was most obvious when rMIF concentration was 200 ng/ml, and the difference was statistically significant compared with L-02 normal hepatocytes ( P<0.05). Conclusion:MIF, ERK1/2 and p-ERK1/2 are highly expressed in HCC tissues and HepG2 HCC cells, suggesting that MIF promotes the occurrence and development of hepatocellular carcinoma through ERK1/2 signaling pathway.

Objective:To explore the performance of the commonly used whole blood C-reactive protein (CRP) detection systems and give related recommendation on the performance requirements of detection systems.Methods:A total of 7 540 venous blood samples from 26 maternal, child and children′s hospitals were collected to conduct this multi-center study on the analytical performance of 5 commonly used whole blood CRP detection systems from March to April in 2019. The blank check, carryover, repeatability, intermediate precision, linearity, sample stability, influence of hematocrit/triglyceride/bilirubin, comparison with SIEMENS specific protein analyzer and trueness were evaluated. The 5 systems included BC-5390CRP autohematology analyzer, AstepPLUS specific protein analyzer, Ottoman-1000 Automated Specific Protein POCT Workstation, i-CHROMA Immunofluorometer equipment Reader and Orion QuikRead go detecting instrument. The 5 systems were labeled as a, b, c, d and e randomly.Results:Within the 5 systems, all values of blank check were less than 1.00 mg/L, the carryovers were lower than 1.00%. The repeatability of different ranges of CRP concentrations including 3.00-10.00, 10.00-30.00 and>30.00 mg/L were less than 10.00%, 6.00% and 5.00%, respectively, and the intermediate precision was less than 10.00%. The linearity correlation coefficients of the 5 systems were all above 0.975, while the slope was within 0.950-1.050. Whole blood samples were stable within 72 hours both at room temperature (18-25 ℃) and refrigerated temperature (2-8 ℃). The CRP results were rarely influenced by high triglyceride or bilirubin, except for the immmunoturbidimetric test based on microparticles coated with anti-human CRP F(ab) 2 fragments. When triglyceride was less than 15.46 mmol/L, the deviation of CRP was less than 10.00%. When bilirubin was less than 345.47 μmol/L, the deviation of CRP was less than 10.00%. CRP was more susceptible to Hct on the systems without Hct correction. The deviation of CRP between different Hct dilution concentration and 40% dilution concentration can reach as high as 67.48%. The correlation coefficients ( r) of 5 systems were all more than 0.975 in the range of 0-300.00 mg/L compared with Siemens specific protein analyzer. All systems passed the trueness verification using the samples with specified values of 12.89 and 30.60 mg/L. Conclusion:The performance of 5 systems can basically meet the clinical needs, but it is suggested that the whole blood CRP detection system without automatic Hct correction should be modified manually.

Objective To investigate intradermal injection and subcutaneous injection of Gadodiamide MR lymphangiography (MRL) in evaluation on the central conducting lymphatics in rabbits.Methods T1 weighted three-dimendional fast low angle shot (fl3d) sequence was used before and after administration of Gadodiamide to undertake MRL.Gadodiamide was administed intrademally in the left and right footpad in 8 rabbits,and then undertake MRL in 1.5T MRI system (intradermal injection group).Three days later,Gadodiamide was administered subcutaneously in the left and right footpad in 8 rabbits,and MRL were underdone with the same sequence (subcutaneous injection group).The degree of contrast enhancement within the lumbar lymphatic trunk and thoracic duct were evaluated using a 3-point scoring system.Results Intradermal injection group showed the popliteal lymph nodes,sacral lymph node and inferior aortic lymph node enhanced obviously in all 8 rabbits,and the enhancement duration time was about 30 90 mini the lumbar lymphatic trunk and cisterna chyli were detected in 6 rabbits,and part of the thoracic duct were detected in 5 rabbits.Subcutaneous injection group showed the popliteal lymph nodes enhanced obviously in all 8 rabbits,but the lumbar lymphatic trunk and the thoracic duct were not detected.The score of contrast enhancement within the lymphatic system of intradermal injection group and subcutaneous injection group had statistical difference (t=100.00,P=0.0002).Conclusion MRL with intradermal injection Gadodiamide are better than subcutaneous injection in evaluation of the lumbar lymphatic trunk.

The rules of acupoint selection for diabetic peripheral neuropathy (DPN) based on data mining technology are analyzed. Literature regarding acupuncture for DPN published in core journals from January of 2005 to December of 2014 was retrieved in CNKI database and Wanfang database to establish a database of acupuncture prescriptions, which were included into the TCM Inheritance Platform software (Version 2.0). The data mining methods, including rule analysis, revised mutual information, etc. were applied to analyze the acupuncture prescriptions and the frequency of each acupoint was calculated. The results indicated Zusanli (ST 36), Sanyinjiao (SP 6), Quchi (LI 11), Yanglingquan (GB 34), Hegu (LI 4), Shenshu (BL 23), etc. were mostly selected in acupuncture for DPN; in addition, 19 acupoint combinations were acquired. It is concluded that acupuncture for DPN applied reinforcing methods as main rules, accompanied with promoting blood andcirculation; the acupoint selection focused onmeridians and back-points.

OBJECTIVETo investigate the changes of cytokines in induced sputum at different stages of silicosis patients.

METHODSA total of 200 workers from one of the Shandong Province gold mine were chosen as object of observation. Among which 40 patients at silicosis stage I and 40 patients at silicosis stage II were divided into silicosis observed object group, silicosis stage I group, silicosis stage II group, and another 80 workers exposed to silica dust without suffering from silicotic Clinical symptoms, however, were chosen as group of dust exposed, and 40 logistical workers without being exposed and history of silicosis's illness were chosen as control group. And ask their basic information by questionnaire. Then, spray-inhalation the induced sputum and apply the ELISA to assess the level of tumor necrosis factor (TNF), interleukin (IL), macrophage inflammatory protein-1 (MIP-1α), monocyte chemotactic factor-1 (MCP-1), metalloproteinases (MMP), transforming growth factor-β (TGF-β), platelet derived growth factor (PDGF) in induced sputum from subjects.

RESULTSThe level of TGF-β [(901.60 ± 30.09) ng/L] in the induced sputumof patients in silicosis stage I group is lower than that in the observed object group [(913.02 ± 20.51) ng/L], and the level of MMP-9 [(212.49 ± 5.97) ng/L], MCP-1 [(129.91 ± 4.30) ng/L] has various degrees of increase than that in control group, observed object group and dust exposed group. All the differences have statistical significances (P < 0.05). The level of TNF-α [(85.76 ± 3.78) ng/L] in the induced sputum of patients in silicosis stage I group reaches the maximum, there are significant differences comparing with that level in the silica dust exposure group and the control group, whose differences are statistically significant (P < 0.05). Compared with the control group, the level of MMP-2 (427.95 ± 23.64) in the induced sputum of patients in silicosis stage I group has increased, whose differences also have statically significant (P < 0.05). Compared with the control group, silica dust exposed group, the observation group of objects, the pneumosilicosis patients of IL-16 in induced sputum IL-16 (21.40 ± 9.24) decreased, the content of PDGF [(5.96 ± 0.51) ng/L], MMP-2 [(447.86 ± 27.10) ng/L], MMP-9 [(223.91 ± 12.28) ng/L], MCP-1 [(122.87 ± 6.08) ng/L] increased, the differences are statistically significant (P < 0.05).

CONCLUSIONAs silicosis biomarkers, TNF-alpha, TGF-beta, IL-16, PDGF, MMP-2, MMP-9 and MCP-1 have certain significance, further suggesting that early detection rate of patients with silicosis can be improved by employing the multiple indexes discriminate equation.

Biomarkers ; metabolism ; Case-Control Studies ; Chemokine CCL2 ; metabolism ; Chemokine CCL3 ; metabolism ; Cytokines ; metabolism ; Discriminant Analysis ; Dust ; Humans ; Interleukin-16 ; metabolism ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Platelet-Derived Growth Factor ; metabolism ; Silicosis ; diagnosis ; Sputum ; chemistry ; Transforming Growth Factor beta ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To discuss the effect and safety about large dosage of tilofiban injection into coronary artery in patients with ST-segment elevated myocardial infarction (STEMI) undergoing primary percutaneous coronary intervention (PCI). Methods A prospective study was conducted. Two hundred and eighteen patients with STEMI admitted into Cardiology Department of Taizhou Central Hospital were enrolled. According to the difference in dosage, they were divided into a large dosage tilofiban group (102 cases) and a routine dosage tilofiban group (116 cases). In both groups, they received the injection of load dosage of tilofiban into coronary artery during they underwent primary PCI, the load dosage being 25μg/kg in the large dosage group, and 10μg/kg in the routine dosage group. Afterwards, the dosage was kept on 0.15μg·kg-1·min-1 in both groups lasting for 18-24 hours. The flow of thrombolysis in myocardial infarction (TIMI) immediately after PCI, the return of ST-segment after operation for 2 hours, the rate of bleeding events, the rate of major adverse cardiac event [MACE, including death, re-infarction and target vessel revascularization (TVR)] and prognosis after operation for 30 days were observed. Results The ratios of the immediate reflow of TIMI 3 grade after operation and the return of ST-segment after operation for 2 hours in the large dosage tirofiban group were higher than those in the routine dosage tirofiban group [the ratio of the reflow of TIMI 3 grade:92.16%(94/102) vs. 81.90%(95/116), the ratio of the return of ST-segment after operation for 2 hours:89.22%(91/102) vs. 73.28%(85/116), both P < 0.05]. The ratios of re-infarction, TVR and the total MACE in 30 days after operation in large dosage tirofiban group were lower than those in the routine dosage tirofiban group [re-infarction: 0.98% (1/102) vs. 2.59% (3/116), TVR: 0.98% (1/102) vs. 2.59% (3/116), total MACE: 1.96% (2/102) vs. 6.03% (7/116), all P < 0.05]. There were no statistically significant differences in mortality and the bleeding events between the large dosage tirofiban group and routine dosage tirofiban group [mortality:0 (0/102) vs. 0.86%(1/116), bleeding events:1.96%(2/102) vs. 0.86%(1/116), both P>0.05]. Conclusion The injection of a large dosage of tilofiban into a coronary artery in patients with STEMI undergoing primary PCI is an effective and safe method to allow them to get more clinical benefits.