Objective To investigate the effect of age on the incidence of cirrhosis and liver cancer in patients with chronic hepatitis B.Methods 279 patients with chronic hepatitis B were divided into the senior group and the younger group according to the age of the patients.The cumulative incidence of cirrhosis and liver cancer during 25 years of follow-up was calculated by using SPSS and R language through the long-term follow-up of HIS system,and the risk factors were analyzed by multivariate logistic regression.Results During follow-up,24 cases developed cirrhosis and 12 cases developed liver cancer.The cumulative incidence of liver cirrhosis was 1.5%,2.1%,5.4%,11.6%and 15.5%in the 5-year,10-year,15-year,20-year and 25-year group,and 5.5%,9.8%,22.9%,29.0%and 52.1%in the elderly,respectively.The difference between the younger age group and senior age group was statistically significant(P<0.001).A total of 2 risk factors(age and follow-up time)were included in the regression model.Two cases in the younger group developed into liver cancer after 17 and 21 years of follow-up,respectively.The cumulative incidence rates at 5,10,15,20 and 25 years were 1.8%,3.8%,18.5%,21.8%and 26.7%.A total of five factors(initial age,HBV-DNA load,HBV-DNA turned negative before the end-point,follow-up time,and sex)were included in the regression model.Conclusions The incidence of cirrhosis and liver cancer in CHB patients aged≥40 years,especially in male patients,is significantly higher than younger CHB patients.Timely initiation of antiviral therapy can delay disease progression and reduce the incidence of termi-nal liver disease.Whether antiviral therapy should be initiated for people aged 30 to 40 years remains to be studied.

Objective:To explore the effect and underlying mechanism of casein kinase 2 interacting protein-1 (CKIP-1) on hepatocyte apoptosis in nonalcoholic fatty liver disease (NAFLD).Methods:Experimental study. An NAFLD cell model was established by inducing human hepatoma cell line, HepG 2 cells, with oleic acid (OA). Flag-CKIP-1 expression vector and shRNA-CKIP-1 were transfected into HepG 2 cells. Flow cytometry was used to detect the effect of CKIP-1 on the activity and apoptosis of NAFLD hepatocytes. The levels of apoptosis-related proteins were detected by Western blot. CKIP-1 knockout mice in C57BL/6 back-ground were fed with either standard or high-fat diet for 8 weeks. Apoptosis-related signal proteins in NAFLD hepatocytes were detected by immunohistochemistry. Results:After CKIP-1 was transfected into HepG 2 cells, the degree of OA induced cell liposis was significantly reduced ( P<0.05). Annexin V-FITC/PI flow cytometry showed that CKIP-1 reduced the apoptosis of steatotic hepatocytes. Overexpression of CKIP-1 could significantly inhibit the expression of caspase-3 and caspase-9 and increase the expression of Bcl-2/Bax ( P<0.05). Knockdown of CKIP-1 could increase the expression of caspase-3 and caspase-9 ( P<0.05). CKIP-1 knockout could further increase the expression of caspase-3 and caspase-9 in NAFLD mice ( P<0.01, P<0.05), and further decrease the expression of Bcl-2/Bax ( P<0.05). Conclusion:CKIP-1 inhibited the apoptosis of steatotic hepatocytes by up-regulating the expression of apoptosis inhibitor gene, Bcl-2/Bax, and affecting the proteases, caspase-3 and caspase-9.

Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 β-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarS_cyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarS_cyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarS_cyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarS_cyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.
Humans ; Histidine Kinase/metabolism* ; Fusobacterium nucleatum/metabolism* ; Automobiles ; Protein Kinases/genetics* ; Escherichia coli/metabolism* ; Colorectal Neoplasms

Based on polyketide syntheses gene (PKS) and non-ribosomal peptide synthetases gene (NRPS), one strain with high anti-pathogenic activity was screened from 77 strains isolated from Arctic marine sediments and identified. By optimizing the composition of culture medium and fermentation conditions, the production of this strain's active metabolites was improved and the main metabolites were identified by HRMS, ¹H NMR and ¹³C NMR. The antibacterial spectrum of the main metabolites and the effect of the metabolites on cucumber Fusarium wilt were also determined. The results showed that the strain was Bacillus velezensis and it showed growth promoting effect on plants. When the strain was cultured in 5 g/L maltose, 10 g/L tryptone, 10 g/L sodium chloride, at 30 ℃, 150 r/min for 60 h, the diameter of the inhibition zone increased from (16.23±0.42) to (24.42±0.57) mm. The metabolites of this strain mainly contain macrolide compound macrolactin A, which has antagonistic effect on a variety of pathogenic bacteria and fungi. Cucumber seedling experiments showed that the metabolites of this strain had a protective effect on cucumber Fusarium wilt, and showed a good potential for development and application as a biocontrol agent.
Polyketides/pharmacology* ; Fungi ; Bacteria ; Fusarium/genetics* ; Anti-Bacterial Agents/pharmacology* ; Peptide Synthases/genetics*

Cyclic lipopeptide has extensive application prospect in the field of medicine due to its unique chemical structure and biological activity. This study aims to obtain high purity of cyclic lipopeptide monomer from Bacillus amyloliquefaciems strain Q-426, and illuminate preliminary antitumor mechanism of C-15 Bacillomycin D and C-16 Bacillomycin D. Firstly, crude cyclic lipopeptide solution was prepared by two-steps purification of acid precipitation and double-resins chromatography. In order to obtain purer product preparative HPLC was utilized to separate and purify cyclic lipopeptide. Component 1 and component 2 were detected as C-15 Bacillomycin D and C-16 Bacillomycin D by HPLC-MS and ESI-MS/MS. Secondly, the effect of C-15 Bacillomycin D, C-16 Bacillomycin D and their mixture (1:1, mol:mol) on cell proliferation was measured using human cancer cells (Hela, MG, Hep-G2 and HT-29). The cyclic peptide showed a dose dependent manner on the cell proliferation inhibition of Hela and MG cells. Finally, the results of the scratch wound healing assay and FACS analysis revealed that C-16 Bacillomycin D can effectively influence the cells migration and the cells treated with C-16 Bacillomycin D showed typical apoptotic morphology with the increase of drug concentration in the early apoptosis, late apoptosis percentage increased, and G₀G₁ arrest was induced significantly.

Objective To analyze the status of sleep disorders in patients with functional gastrointestinal disease (FGID) and its relation with symptom characteristics .Methods From January to December 2014 ,questionnaire was carried out in FGID patients who met the Rome Ⅲ criteria and visited the outpatient department of gastroenterology at six third-level general hospitals in Tianjin City to assess the severity of symptoms ,sleep quality (Pittsburgh sleep quality index ,PSQI) ,and psychological state (anxiety and depression) .Chi-square test and Mann-Whitney rank sum test were performed for statistical analysis .Results Among 931 patients with FGID ,651 (69 .92％ ) patients had sleep disorders and 280 (30 .08％ ) patients had no sleep disorders .Among 828 patients with functional dyspepsia (FD) ,360 (43 .48％ ) patients had sleep disorders complicated with and depression .Among 292 patients with irritable bowel syndrome (IBS ) , 138 (47 .26％ ) had sleep disorders complicated with anxiety and depression .Among 618 patients with FD complicated with sleep disorders , 70 (11 .33％ ) patients overlapped with IBS ;among 210 patients with FD ,but without sleep disorder ,11 (5 .24％ ) patients overlapped with IBS and the percentage of the former was higher than the latter ,and the difference was statistically significant (χ2 =6 .580 , P=0 .01) .The proportion of lower abdominal pain ,sheep fecal or hard stool ,laborious defecation or incomplete defecation in FGID patients without sleep disorder were 22 .14％ (62/280) ,11 .79％ (33/280) ,19 .29％ (54/280) and 27 .86％ (78/280) ,respectively ;which were lower than those of FGID patients with sleep disorders (36 .10％ (235/651) ,21 .20％ (138/651) ,32 .41％(211/651) and 44 .39％ (289/651));and the differences were statistically significant (χ2 =17 .552 ,11 .569 , 16 .566 and 22 .419;all P<0 .01) .FGID patients with sleep disorders have more severe symptoms such as lower abdominal pain , lower abdominal discomfort (non-pain ) , sheep fecal or hard stool , laborious defecation incomplete defecation , and urgency than FGID patients without sleep disorders ;and the differences were statistically significant (Z= -4 .423 ,-1 .973 ,-3 .360 ,-4 .467 ,-4 .550 and -2 .420 ;all P<0 .05) . Conclusions Sleep disorders ,anxiety and depression often coexist in patients with FGID .Sleep disorders are closely related with lower gastrointestinal symptoms in patients with FGID .

Objective To investigate a sonic wave device including micro-oscillator and microphone for real-time monitoring the situation of the esophageal motility of lower esophageal sphincter (LES). Methods A micro-oscillator that can actively send sound waves was located at one side of LES, and a microphone located at the other side can be to capture the sound generated from the oscillator. This device was tested in vitro by utilizing a LES simulator in quiet and noisy environment, as well as in vivo in a pilot model corresponding to moments A-D. Results In the in vitro test, the sound can be captured by the microphone, and its power strength was correlated with closing and opening of LES. The measurements showed statistically significant Pearson correlation coefficients. In the in vivo test, the LES was open and closed by an endoscope, which was monitored in real-time by another endoscope inserted into the distal stomach from the duodenum. The sound power strength corresponding to moments A-D were (-89.63 ± 0.79) dB, (-84.74 ± 0.97) dB, (-82.37 ± 1.93) dB and (-89.86 ± 0.75) dB. Conclusion The LES dynamics can be monitored by the acoustic interrogation device, which has the potential to become an effective minimally-invasive technique for studying LES dysfunction.

Objective To evaluate the results of different methods in removing the root canal gutta percha.Methods Select 60 man-dibular premolars with step-back techinique during root canal prepartion,and the mandibular premolars were stored for 2 weeks after AH-plus paste and gutta percha cold lateral condensation.And then they were randomly divided into 3 groups to remove the root canal filling.Group A were given manually cleaning and shaping with stainless steel instruments combined with chloroform.Group B were given Pro Taper nickel ti-tanium system.Group C were given Pro Taper nickel titanium system combined with microscopic ultrasound technology.The completion time of the 3 groups were recorded and the root canal wall cleanliness of the 3 groups were carefully observed.Results The competition time and amount of debris of apical foramen in group C were both significantly less than those of group A and group B,with statistically significant difference (P <0.05).And the cleanliness of root canal wall of group A was significantly better than that of group A and group B (P <0.05). Conclusion During the root canal retreatment,mechanically use Pro Taper nickel titanium system combined with microscopic ultrasound technology can improve the work efficiency and the cleanliness of root canal wall.

Objective To study the effects of tonifying kidney therapy on pathology in chronic hepatitis B virus carriers.MethodsWith the multi-center, randomized, double-blinded and placebo-controlled methods, 600 cases of chronic hepatitis B virus carriers were divided intoBushen Qingtou group,Bushen Jianpi group and control group, 200 cases in each group, and were treated withBushen Qingtou prescription,Bushen Jianpi prescription and placebo prescription respectively for 52 weeks. The pathological changes of the liver biopsy were observed by liver biopsy examination before and after treatment. Inflammatory active degree and fibrosis were scored with Knodell HAI and Ishak.Results The number of decreasing more than 2 points on Knodell HAI inBushen Qingtou group,Bushen Jianpi group and control group was 21, 18, and 6 respectively (P<0.05); the number of increasing more than 2 points on Knodell HAI in three groups was 3, 2, and 8 respectively (P<0.05). The curative effect ofBushen Qingtou prescription,Bushen Jianpi prescription were significantly obviously better than control group, without statistical significnce between the two treatment groups (P>0.05). The number of decreasing more than 1 points on Ishak in Bushen Qingtou group,Bushen Jianpi group, and control group was 13, 12, and 9 respectively (P>0.05); the number of increasing more than 1 points on Ishak inBushen Qingtou group,Bushen Jianpi group and control group was 8, 3, and 11 respectively, with statistical significance betweenBushen Jianpi group and controlled group (P<0.01), without statistical significance betweenBushen Qingtou group and control group (P>0.05), which meantBushenJianpi prescription could prevent the deterioration of liver tissue fibrosis more significantly than placebo prescription did. ConclusionTonifying kidney therapy, includingBushen Qingtou prescription andBushen Jianpi prescription, can inhibit the inflammatory activity and slow down the fibrosis progression of the chronic HBV carriers.

Objective To discuss the short-term curative effect and the safety of percutaneous vertebroplasty using high viscosity bone cement for the treatment of severe osteoporotic vertebral body compression fractures. Methods The clinical data of 100 patients with severe osteoporotic vertebral body compression fractures (compression degree>70%), who received percutaneous vertebroplasty by using high viscosity bone cement during the period from December 2010 to May 2013, were retrospectively analyzed. All the patients were followed up for at least one month. Both preoperative and postoperative visual analogue scale (VAS) and quality of life (QOL) scores, as well as the incidence of bone cement leakage, were recorded and the results were used to evaluate the curative effect and the safety of percutaneous vertebroplasty for severe osteoporotic vertebral body compression fractures. Results One week after the treatment, significant pain relief was obtained in 92 patients (92%), VAS scores decreased from preoperative (7.0 ±1.2) to postoperative (2.0±1.5), and QOL scores increased from preoperative (30±5.0) to postoperative (80±18.0);the differences were statistically significant (P<0.01). One month after the treatment, significant pain relief was seen in 91 patients (91%), VAS scores decreased from preoperative (7.0 ±1.2) to postoperative (1.5 ±1.0) and QOL scores increased from preoperative (30±5.0) to postoperative (80±15.0); the differences were statistically significant (P<0.01). No statistically significant differences in VAS scores and QOL scores existed between the data determined at one week after the treatment and the data determined at one month after the treatment (P>0.05). Leakage of bone cement was observed in 40 patients (40%), resulting no severe neurological symptoms; among the 40 patients, intervertebral disc leakage at above and below the vertebral body was detected in 28 patients (70%), vertebral anterior edge leakage was observed in 11 patients (27.5%) and vertebral posterior edge leakage was seen in one patient (2.5%). Conclusion For the treatment of severe osteoporotic vertebral body compression fractures, percutaneous vertebroplasty by using high viscosity bone cement is safe and effective.