Objective:To explore the flora characteristics and differences of esophageal tissues between elderly esophageal squamous cell carcinoma (ESCC) patients and young and middle-aged ESCC patients, so as to assist in studying the potential biomarkers of elderly ESCC patients.Methods:In this study, a retrospective study was adopted. 72 ESCC patients diagnosed in Taihe Hospital, Shiyan City, Hubei Province from July 2018 to July 2019 were selected, including 49 patients in the elderly group (≥ 60 years old, 40 males and 9 females), 23 patients in the young and middle-aged group (<60 years old, 21 males and 2 females). In the same period, 20 healthy persons without abnormal gastroscopy in endoscopy center were selected as the control group (aged 35-78 years old, median age 57 years old, 16 males and 4 females). The genomic DNA was extracted from the affected esophageal tissues of patients with ESCC and the middle esophageal samples of the control group. The V4 hypervariable region of bacterial 16SrRNA gene sequence was amplified. Illumina HiSeq sequencing technology was adopted. The flora characteristics of elderly, young and middle-aged ESCC patients was compared and analyzed. QIIME and Rstudio software were used to analyze the sequence data, and nonparametric Kruskal-Wallis test or Wilcoxon rank sum test were used for statistical methods.Results:Shannon index [5.17 (4.53, 5.95) vs. 4.79 (3.74, 5.97)], Simpson index [0.94 (0.91, 0.96) vs. 0.92 (0.83, 0.96)] and Chao1 index [343.55 (259.76, 570.59) vs. 329.16 (268.88, 648.00)] were similar in flora of two groups, and there was no significant difference ( Z=-0.791, -1.057, -0.380, all P>0.05). There was no significant difference in β-diversity between the elderly group and the young and middle-aged group (PC1=19.14%, PC2=6.95%, PPC1=0.67, PPC2=0.42). At the phyla level, the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria in the young and middle-aged group, while the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Fusobacteria and Actinobacteria in the elderly group; the significant difference between the two groups was Fusobacteria ( Q=0.596, P<0.05). At the genus level, the top 5 genera in the young and middle-aged group in abundance were as follows: Prevotella, Bacteroides, Streptococcus, Selenomonas and Veillonella. In the elderly group, Prevotella, Bacteroides, Streptococcus, Selenomonas and Haemophilus were the top 5 in abundance, and there were significant difference in Fusobacterium between the two groups ( Q=0.938, P<0.05). PICRUSt function prediction showed that the abundance of Aminoacyl.tRNA.biosynthesis, Nucleotide.excision.repair, RNA.polymerase, Ribosome, Clavulanic.acid.biosynthesis, Photosynthesis and Photosynthesis. proteins in the elderly group were lower than those in the young and middle-aged group (all Q=0.734, P<0.05). Conclusion:There is no significant difference in α-diversity and β-diversity between elderly ESCC patients and young and middle-aged patients, but the abundance of Fusobacterium flora increased.

Objective:To explore the flora characteristics and differences of esophageal tissues between elderly esophageal squamous cell carcinoma (ESCC) patients and young and middle-aged ESCC patients, so as to assist in studying the potential biomarkers of elderly ESCC patients.Methods:In this study, a retrospective study was adopted. 72 ESCC patients diagnosed in Taihe Hospital, Shiyan City, Hubei Province from July 2018 to July 2019 were selected, including 49 patients in the elderly group (≥ 60 years old, 40 males and 9 females), 23 patients in the young and middle-aged group (<60 years old, 21 males and 2 females). In the same period, 20 healthy persons without abnormal gastroscopy in endoscopy center were selected as the control group (aged 35-78 years old, median age 57 years old, 16 males and 4 females). The genomic DNA was extracted from the affected esophageal tissues of patients with ESCC and the middle esophageal samples of the control group. The V4 hypervariable region of bacterial 16SrRNA gene sequence was amplified. Illumina HiSeq sequencing technology was adopted. The flora characteristics of elderly, young and middle-aged ESCC patients was compared and analyzed. QIIME and Rstudio software were used to analyze the sequence data, and nonparametric Kruskal-Wallis test or Wilcoxon rank sum test were used for statistical methods.Results:Shannon index [5.17 (4.53, 5.95) vs. 4.79 (3.74, 5.97)], Simpson index [0.94 (0.91, 0.96) vs. 0.92 (0.83, 0.96)] and Chao1 index [343.55 (259.76, 570.59) vs. 329.16 (268.88, 648.00)] were similar in flora of two groups, and there was no significant difference ( Z=-0.791, -1.057, -0.380, all P>0.05). There was no significant difference in β-diversity between the elderly group and the young and middle-aged group (PC1=19.14%, PC2=6.95%, PPC1=0.67, PPC2=0.42). At the phyla level, the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Actinobacteria and Fusobacteria in the young and middle-aged group, while the top 5 phyla in abundance were as follows: Firmicutes, Bacteroidetes, Proteobacteria, Fusobacteria and Actinobacteria in the elderly group; the significant difference between the two groups was Fusobacteria ( Q=0.596, P<0.05). At the genus level, the top 5 genera in the young and middle-aged group in abundance were as follows: Prevotella, Bacteroides, Streptococcus, Selenomonas and Veillonella. In the elderly group, Prevotella, Bacteroides, Streptococcus, Selenomonas and Haemophilus were the top 5 in abundance, and there were significant difference in Fusobacterium between the two groups ( Q=0.938, P<0.05). PICRUSt function prediction showed that the abundance of Aminoacyl.tRNA.biosynthesis, Nucleotide.excision.repair, RNA.polymerase, Ribosome, Clavulanic.acid.biosynthesis, Photosynthesis and Photosynthesis. proteins in the elderly group were lower than those in the young and middle-aged group (all Q=0.734, P<0.05). Conclusion:There is no significant difference in α-diversity and β-diversity between elderly ESCC patients and young and middle-aged patients, but the abundance of Fusobacterium flora increased.

Objective: To examine the correlation between the promoter methylation of Sterol regulatory-element binding protein-2 (SREBP-2) and miR-33a expression as well as serum markers in patients with coronary artery disease (CAD). Methods: The case-control study. 100 participants who underwent coronary angiography from August 2017 to April 2018 in TaiheHospital, Hubei University of Medicine, were recruited in this study.The methylation level of two fragments, including 12 CpG sites in the promoter region of SREBP-2, have been detected by pyrosequencing in 50 patients with coronary artery disease (CAD) and 50 non-CAD controls. Serum miR-33a level and a panel of 15 CAD related biomarkers were examined by qPCR and routine biochemistry methods. Results: Methylation level of one CpG site (F1-4 loci) in SREBP-2 promoter region were significant higher in CAD patients than in controls(4.56%±0.70% vs 3.54%±0.72%, t=-3.864, P<0.001); methylation level of F1-4 site was negatively correlates with the serum miR-33a levels and high-density lipoprotein cholesterol (HDL-C) levels(r=-0.318, P=0.001; r=-0.225, P=0.024, respectively). Furthermore, F1-4 hypermethylation was an independent risk factor of CAD, independent of age, gender, histories of hypertension, hyperlipidemia, and diabetes(OR=2.452, 95%CI=1.398-4.299, P=0.002). Conclusion These results suggest that DNA methylation and miRNA might cooperate to regulate the lipid metabolism in CAD.

Objective To explore the feasibility and effect of 3D modeling and printing technology in constructing bone fracture models and assisting clinical teaching at the department of traumatic orthope-dics. Methods CT scan images of bone fractures were reconstructed by Mimics software. The digital 3D bone fracture models were constructed and the interactive multimedia teaching videos were output. More-over, all bone fracture models were printed by using fusion deposition modeling (FDM). At the end of the teaching course, a questionnaire survey was conducted to evaluate the teaching effect. Results The digital models of common bone fractures at the department of traumatic orthopedics were established, and the in-teractive multimedia teaching videos were output. A traumatic orthopedic teaching model with a 1∶1 scale was printed out. The questionnaire survey indicated that the application of 3D modeling and printing tech-nology to build bone fracture model with PPT teaching can obviously improve students' understanding and mastery of relevant theoretical knowledge. They helped students better remember the type of bone fractures and how to choose the correct internal fixation methods. The teaching effect was satisfactory. Conclusions 3D modeling and printing technology was applied to build bone fracture models to assist clinical teaching at the department of traumatic orthopedics. It was found that the printed 3D bone fracture models can stimulate students' enthusiasm for learning and improve their learning effect. This method has good application value.

Objective To investigate the clinical values of detection of plasma miRNA-544a (miR-544a) expression level in the diagnosis and treatment of lung cancer.Methods A total of 110 patients diagnosed with lung cancer in Ⅰ-Ⅱ stage from June 2017 to March 2018 in Shiyan Taihe Hospital were collected,and 35 patients with benign pulmonary nodules and 64 healthy people were also collected as the controls.Realtime quantitative polymerase chain reaction (RT-qPCR) was used to detect the level of miR-S44a in plasma of 64 healthy people,110 patients with lung cancer (50 newly diagnosed patients without anti-tumor treatment,33 patients one week after radical resection,27 patients after one week chemotherapy with the same dose) and 35 patients with benign pulmonary nodules.Of the 50 newly diagnosed patients,42 cases were non-small cell lung cancer and 8 cases were small cell lung cancer.The plasma expression level of miR-544a in each group was compared by using Mann-Whitney U test,and the efficacy of miR-544a in the diagnosis of lung cancer was evaluated by the receiver operationg characteristic (ROC) curve and the area under the curve (AUC).Results The plasma expression levels of miR-544a in the newly diagnosed untreated lung cancer group,one week after operation group,one week chemotherapy group,healthy control group and benign pulmonary nodule group were 1.40 nmol/L (0.55 nmol/L,8.76 nmol/L),33.52 nmol/L (3.64 nmol/L,250.47 nmo/L),8.87 nmol/L (0.68 nmol/L,125.43 nmol/L),0.31 nmol/L (0.17 nmol/L,1.19 nmol/L),1.04 nmol/L (0.31 nmol/L,4.62 nmol/L),respectively,and the differences between untreated lung cancer group and the other 4 groups were statistically significant (Z =-4.483,P ＜ 0.001;Z =-4.274,P ＜ 0.001;Z =-2.562,P =0.01;Z =-2.152,P =0.031).The expression levels of miR-544a in non-small cell lung cancer group and small cell lung cancer group were 1.40 nmol/L (0.66 nmol/L,8.76 nmol/L) and 1.37 nmol/L (0.26 nmol/L,36.97 nmol/L),respectively.The differences between non-small cell lung cancer group and healthy control group and benign pulmonary nodule group were statistically significant (Z =-4.463,P ＜ 0.001;Z =-2.026,P =0.043).Compared with the healthy people,the AUC of miR-544a for diagnosing the lung cancer was 0.841,the sensitivity was 87.5 ％,and the specificity was 68.0 ％.Compared with the benign pulmonary nodule,the AUC for diagnosing lung cancer was 0.638,the sensitive was 45.7 ％,and the specificity was 80.0 ％.Conclusions The plasma expression level of miR-544a has certain significances in the differential diagnosis of early stage lung cancer and benign pulmonary nodules and healthy people,and it can be used as a potential biomarker for diagnosing early stage lung cancer,especially for the non-small cell lung cancer.The plasma expression of miR-544a is increased after surgery or chemotherapy,suggesting that its expression may be related to the occurrence and development of lung cancer,and miR-544a may become a new target for cancer treatment.

OBJECTIVETo assess the association of methylenetetrahydrofolate reductase (MTHFR) gene 677C to T polymorphism with blood homocysteine (Hcy) level among women of childbearing age from Shiyan area.

METHODSPCR-chip hybridization was used to determine the genotype of MTHFR 677C to T, and a biochemical assay was used to determine the total Hcy level among 428 healthy women of childbearing age. Association of MTHFR 677C to T with total Hcy level was assessed.

RESULTSHeterozygous CT mutation was most common form for the MTHFR 677C to T polymorphisms and amounted for 49.77% among the group, while the CC wild type and homozygous TT mutation respectively accounted for 30.61% and 19.63%. These gave a frequency of 44.51% for the 677T allele. The dominant genotype among different age groups were the CT type. Of note, the proportion of MTHFR 677CC is higher in women above 30 years of age. The distribution of MTHFR 677C to T genotypes has differed significantly among different age groups (P<0.05). Compared with those with wild type alleles, carriers of MTHFR mutations had a higher plasma Hcy level. The genotypic frequencies of MTHFR C677T in Shiyan region differed significantly from those of Sichuan, Hebei, Henan and Shandong (P<0.05) but were similar to those of Jiangsu, Guangdong, Ningxia and Xinjiang.

CONCLUSIONThe distribution of MTHFR C677T polymorphism among women of childbearing age in Shiyan area is influenced by age and is geographically specific and associated with plasma Hcy level. Nearly 50% of women have carried the high risk alleles, for whom folic acid supplementation is crucial for the reduction of birth defect rate.

Objective To determinate the serum level of adiponectin,visfatin and resistin in patients with rheumatoid arthritis (RA) and to explore its role in the development of RA and its clinical significance.Methods Blood samples were collected from 67 cases of RA patients and 65 healthy controls,the serum levels of adiponectin,visfatin and resistin were tested by enzyme linked immunosorbent assay (ELISA).Adipokine levels in different groups were compared using the Mann-Whitney U test.Spearman test and multivariate analysis were used to evaluate the correlation with clinical and laboratory parameters [erythrocyte sedimentation rate (ESR),C reactive protein (CRP),blood platelet (PLT),rheumatoid factor (RF),anti-cyclic citrullinated peptide (CCP),disease activity score (DAS)28].Results The serum level of adiponectin,visfatin and resistin in RA patients were significantly higher than those in healthy controls[8.17(4.51,28.53) μg/ml vs 6.83(4.48,25.32) μg/ml,U=1 663,P=0.019];[6.56(5.34,8.40) ng/L vs 4.24 (3.01,6.65) ng/L,U=762,P=0.001];[10.65 (5.99,20.70) ng/ml vs 6.12 (4.49,9.98) ng/ml,U=1 406.5,P=0.000].The serum level of visfatin was positively correlated with RF in patients with RA (r=0.186,P=0.013),and serum levels of adiponectin,visfatin and resistin were positively correlated with DAS28 (r=0.370,0.263,0.285;P＜0.05).The level of visfatin in RA patients with high activity group was higher than that in the low and moderate activity group [4.37(2.72,7.53)ng/L vs 4.11 (3.11,6.44) ng/L,U=133.5,P=0.019].Conclusion Multivariate analysis has showm that ESR,PLT and DAS28 can significantly affect adiponectin,and CRP can significantly influence resistin.Serum level of adiponectin,visfatin and resistin are elevated and correlate with DAS28,suggesting that they may be involved in the chronic inflammation of RA.

Based on the structure and motion bionic principle of the normal adult fingers, biological characteristics of human hands were analyzed, and a wearable exoskeleton hand function training device for the rehabilitation of stroke patients or patients with hand trauma was designed. This device includes the exoskeleton mechanical structure and the electromyography (EMG) control system. With adjustable mechanism, the device was capable to fit different finger lengths, and by capturing the EMG of the users' contralateral limb, the motion state of the exoskeleton hand was controlled. Then driven by the device, the user's fingers conducting adduction/abduction rehabilitation training was carried out. Finally, the mechanical properties and training effect of the exoskeleton hand were verified through mechanism simulation and the experiments on the experimental prototype of the wearable exoskeleton hand function training device.
Bionics ; instrumentation ; Electromyography ; Exoskeleton Device ; Fingers ; Hand ; Humans ; Motion ; Stroke Rehabilitation

Objective To investigate the expression and significance of CCR9/CCL25 pathway in acute rejection of mouse skin transplantation.Method BALB/c mice and C57BL/6 mice were selected as allogeneic and syngeneic skin graft donors and C57BL/6 as recipients,then established a murine skin transplantation model of acute rejection.Allogeneic transplant recipient mice were injected with an AntiCCL25(2 g,as experimental group) mAb or IgG(2 g,as control group) every other day via tail vein,a total of 10 injections.The transplanted graft was scored visually daily,and then we collected skin graft and spleen from recipient mice of each group at 3,5,7 days after transplantation.HE staining was done to analyze necrosis of skin tissue Confocal and immunohistochemistry were also used to measure CCR9 and CCL25 expression in recipient skin grafts.Result HE staining indicated that there was a widespread inflammatory cell infiltration in the skin from allo-transplantation mice,and CCR9 expression measured by immunohistochemistry and confocal was significantly elevated in the surface of the infiltrated CD3 + T cells from skin grafts tissue and spleen.Neutralization of CCL25 with Anti-CCL25 mAb significantly prolonged allogra,ft survival and markedly reduced inflammation.Conclusion CCR9 was highly expressed in the spleen and skin grafts tissue of allogeneic transplanted mice Neutralization of CCL25 by intravenous injection of Anti-CCL25 monoclonal antibody significantly prolonged skin allograft survival.Our study indicates that CCR9/CCL25 pathway is involved in acute rejection process of skin transplantation model in mice were used as syngeneic and skin grit skin graft donor mice.

A method was developed for the simultaneous determination of 17 characteristic ingredients of plant extracts, including paeoniflorin, hydroxysafflor yellow A, calycosin_7_glucoside ferulic acid, etc. , in hair growth cosmetics using ultra high performance liquid chromatography ( UPLC ) . Different cosmetic samples were extracted by ultrasonic_assisted extraction with the solvent of methanol/water (4∶1, V/V) solution. After demulsified by the addition of appropriate amount of NaCl and high speed centrifugation, the supernatant was transferred and analyzed with UPLC. The separation was conducted on a Waters reversed phase column of ACQUITY UPLC CSH C18(50 mm×2. 1 mm, 1. 7μm), and the mobile phases were methanol and the solution of 0. 05% phosphate in water. The detection was performed with a photodiode_array ( PDA) detector. The linear range was 0 . 2-25 mg/L with correlation coefficients higher than 0 . 999 . The limits of detection were within 0. 3-1. 5 mg/kg, and the limits of quantification were from 1. 0 to 4. 0 mg/kg. The average recoveries of 17 characteristic ingredients were within 93 . 5%-105 . 0%, with the intra_and inter_day precision ( n=6 ) less than 4. 6%. This method was simple, rapid, with good_repeatability, and had been applied to the analysis of real samples.