OBJECTIVE: To investigate the changes in bacterial flora in fecal samples, at the tumor loci and in adjacent mucosa in patients with colorectal cancer (CRC). METHODS: We collected fecal samples from 13 patients with CRC and 20 healthy individuals and tumor and adjacent mucosa samples from 6 CRC patients. The differences in bacterial composition between the fecal and mucosa samples were analyzed with 16S rDNA sequencing and bioinformatics methods. We also detected the total number of bacteria in the feces using flow cytometry, isolated and identified the microorganisms in the fecal and mucosa samples using common bacterial culture media. We further tested the effects of 7 isolated bacterial strains on apoptosis of 3 CRC cell lines using lactate dehydrogenase detection kit. RESULTS: The bacterial α-diversity in the feces of healthy individuals and in adjacent mucosa of CRC patients was significantly higher than that in the feces and tumor mucosa in CRC patients (P < 0.05). Lactobacillaceae is a specific bacteria in the feces, while Escherichia, Enterococcus, and Fusobacterium are specific bacteria in tumor mucosa of CRC patients as compared with healthy individuals. Cell experiment with3 CRC cell lines showed that Bacteroides fragilis isolated from the tumor mucosa of CRC patients produced significant inhibitory effects on cell proliferation (P < 0.0001), while the isolated strain Fusobacterium nucleatum obviously promoted the proliferation of the cell lines (P < 0.001). CONCLUSION The bacterial flora in the feces, tumor mucosa and adjacent mucosa of CRC patients is significantly different from that in the feces of healthy individuals, and the fecal flora of CRC patients can not represent the specific flora of the tumor mucosa. Inhibition of F. nucleatum colonization in the tumor mucosa and promoting B. fragilis colonization may prove beneficial for CRC treatment.
Bacteria ; Colorectal Neoplasms/pathology* ; Feces/microbiology* ; Gastrointestinal Microbiome ; Humans ; Intestinal Mucosa

CY-1-4 is a tryptanthrin derivative exhibiting antitumor activity. The solubility of CY-1-4 was poor and the corresponding mechanism needs further study. To solve this problem, we prepared nanoparticles encapsulated with CY-1-4 (CY-1-4 NPs) by nanoprecipitation method using poly(caprolactone) (PCL) and poly(ethylene glycol)-co-poly(ε-caprolactone) (PEG-PCL) as carriers to improve solubility. We then explored whether CY-1-4 NPs induced B16-F10 cytotoxicity via ferroptosis by determining the effect of CY-1-4 NPs on reactive oxygen (ROS) levels, repairing efficacy of lipid reactive oxygen inhibitor ferrostatin-1 and iron chelator deferoxamine (DFO), and potentiation of protoporphyrin (PPIX) induced B16-F10 cell death. The results showed that nanoparticlated strategy significantly improved solubility of CY-1-4. With the particle size about 116 nm, encapsulating efficacy was about 83% and the drug loading capacity was about 4.80%. Ferroptosis mechanistic studies indicated that CY-1-4 NPs could improve the ROS level in B16-F10 cells, whereas ferrostatin-1 and DFO could partly inhibited the cytotoxicity and PPIX could potentiated the cytotoxicity of CY-1-4 NPs in B16-F10 cells. These results showed that ferroptosis was one of the cell death mechanisms induced by tryptanthrin derivative CY-1-4 nanoparticle.

BACKGROUND:During the percutaneous vertebroplasty, the optimal dose of bone cement that can bring favorable cement dispersion and remodel the biomechanical balance of the fractured vertebrae remains controversial. OBJECTIVE:To investigate the dispersion degree of small dose of bone cement in vertebroplasty. METHODS: In this experiment, 18 sheep selected with the same condition were randomly divided into three groups (group A, group B, group C), 6 in each group. A model of thoracolumbar vertebral compression fracture (T12, L1, L2) was made in each sheep. The injected volume of bone cement in groups A, B, C was 15％, 20％, 25％ of the average volume of adjacent vertebral bodies, respectively. Postoperative CT images were used to evaluate the bone cement dispersion. Dispersion degree of bone cement among the three groups was compared by the Kruskal-Wallis test. RESULTS AND CONCLUSION:There was no statistical difference in the dispersion degree of bone cement among the three groups, and the excellent and good rate of dispersion was over 80％. To conclude, the optimal dose of bone cement injected into the fractured vertebra is 15％ of the average volume of adjacent vertebral bodies, which can achieve good dispersion degree and restore the biomechanical stability of the vertebral body.

Objective To study the expression and significance of microRNA-130a(miR-130a) and miR-130b in cirrhosis' serum.Methods The elbow vein blood of 15 cases of healthy subjects and 15 cases of patients with cirrhosis were collected and were divided into treatment group and control group respectively.Reverse transcription-polymerase chain reaction was used to detect the expression of miR-130a and miR-130b in serum.Results The relative expression of miR-130a in treatment group and control goup were (1.57 ±0.36)and (1.40 ± 0.46) and the relative expression of miR-130b in serum in treatment group and control group were (2.18 ± 0.60) and (1.64 ±0.26).The differences were statistically significant(P ＜0.05 or P ＜0.01).Conclusion The expression level of miR-130a and miR-130b in cirrhosis' serum is significantly higher than those in healthy person.It is likely to be used as a diagnostic marker for cirrhosis.

Objective To study the dynamic changes of cytochrome P4502E1 (CYP2E1) in alcohol-induced liver injury in mice and its sig nificance.Methods Fifty male mice were randomly divided into modelgroup (n =40) and control group (n =10).The model of alcoholic liver injury was established by continuous infusion of 56 ° Erguotou at 10 mL · kg-1 for 4 weeks.At the time of 1,2,3,4 weeks,to take 10 mice,eye blood for serum aspartate aminotransferase (AST) activity determina tion.Followed by cervical dislocation of mice,the number of CYP2E1positive cells in mice liver were detected by immunohistochemical SP method.Results The activity of hepatic AST enzyme in model group were (126 ±24),(967 ±30),(1010 ±35) and (206 ±23) U · L-1 in 1st,2nd,3rd and 4th weeks,respectively.Compared with the control group (112 ±22) U · L-1,the activity of hepatic AST enzyme in the model group increased continuouslyfrom 1st to 3rd weeks and reached the peak at 3rd week with significantly (P ＜ 0.01),which was decreased at the 4th week,but still higher than the control group with significantly (P ＜0.05).The number of CYP2E1 positive cells per square millimeter were (3.2 ±0.8),(8.4 ± 1.1),(13.2 ± 1.3),and (4.6±0.8) cell · mm-2 in 1st,2nd,3rd and 4th weeks.Compared with the control group (2.8 ±0.5) cell ·mm-2,the number were obviously increased in 1st,2nd and 3rd weeks and represented statistically significant (P ＜0.01)while the number of the 4th week decreased but still with statistically significant(P ＜0.05).Conclusion The expression of CYP2E1 was increased first and then decreased in alcohol-induced liver injury model,which was consistent with the trend of serum AST enzyme activity.The dynamic changes in expression may be associated with damage and repair of the hver.

OBJECTIVETo summarize the prognostic factors of stage 3 colorectal cancer.

METHODSThe clinical data of 433 patients with stage 3 colorectal cancer who were admitted to our hospital from January 2005 to December 2008 for radical surgery and adjuvant chemotherapy were retrospectively analyzed. Relationship of their clinicopathologic features and treatment with the prognosis were analyzed.

RESULTSOf these 433 stage 3 patients,the mean disease-free survival was (72.37 ± 2.11) months and mean overall survival was (79.91 ± 2.02) months; however, the median survival times were not reached. The 1-,3-, and 5-year disease-free survival rate were 86.8%,77.9%, and 57.0% and the overall survival rate were 91.5%,75.1%, and 63.3%. Multivariate COX regression analysis displayed that intestine obstruction before surgery, complications after surgery,tumor location,positive surgical margin, neural cell infiltration,vessel cancer embolus, TNM stage, lymph node ratio, adjuvant chemotherapy regimens, and chemotherapy duration were the independent factors affecting disease-free and overall survivals in patients with stage 3 colorectal cancer. The efficacies of FOLFOX and XELOX regimens were significantly correlated with patient's age, complications,tumor location,and chemotherapy duration.

CONCLUSIONSComplications,tumor location, TNM stage, and positive surgical margin are the independent prognostic factors of stage 3 colorectal cancer. FOLFOX and XELOX regimen can remarkably improve prognosis,and a longer duration of chemotherapy can achieve better survival.

Antineoplastic Combined Chemotherapy Protocols ; Chemotherapy, Adjuvant ; Colorectal Neoplasms ; Deoxycytidine ; analogs & derivatives ; Disease-Free Survival ; Fluorouracil ; analogs & derivatives ; Humans ; Lymph Nodes ; Neoplasm Staging ; Prognosis ; Retrospective Studies ; Survival Rate

OBJECTIVETo establish an ideal CCl4 drug-induced liver injury model in vitro.

METHODTraditional method and improved method were adopted for preparing CCl4 injury liquid and drug-induced human liver HepG2 cell injury. Cell morphological change was observed under a bright-field microscope. The level of alanine aminotransferase (ALT) in supernatant was detected by biochemical method. 4-Methyl-tetrazolium (MTT) chromatometry was adopted for determining cell activity.

RESULTThe improved method showed better CCl4-induced injury effect than the traditional method. With the increase in the concentration of CCl4 injury liquid, the ALT level significantly increased, whereas the cell activity notably decreased. Particularly, 70% CCl4 injury liquid use for 4 hours could achieve the best injury effect.

CONCLUSIONThe improved method could be used to establish an ideal CCl4 drug-induced liver injury model in vitro, which can lay foundation for further in vitro studies.

Alanine Transaminase ; metabolism ; Aspartate Aminotransferases ; metabolism ; Carbon Tetrachloride ; adverse effects ; Chemical and Drug Induced Liver Injury ; enzymology ; etiology ; Hep G2 Cells ; Humans ; Liver ; drug effects ; enzymology ; injuries ; Models, Biological ; Superoxide Dismutase ; metabolism

OBJECTIVETo explore the diagnostic value of combined modified Alvarado scores (MAS) and computed tomography imaging in the pathological types of acute appendicitis in adults.

METHODSClinical data of a total of 396 adult patients with acute appendicitis confirmed by surgery and pathology were analyzed retrospectively from June 2007 to July 2010. Case-control study was used to investigate the MAS. CT signs were studied in 115 patients who underwent preoperative CT scan. Univariable analysis was performed using each indicator among different pathological types. Discriminant classification was formed by applying significant variables identified from univariable analysis and a Fisher discriminant function was created.

RESULTSTwenty three variables were statistically significant among different pathological types after univariable analysis(P<0.05) and were selected for discriminant analysis. Six variables including temperature(X1), leucocyte count(X2), the proportion of neutrophil(X3), MAS points(X4), periappendiceal fat stranding(X5), and extraluminal air(X6) were enrolled. The discriminant function equation was Y1=0.012X1+0.041X2+0.069X3-0.039X4+2.653X5+1.418X6, Y2=0.327X1+0.041X2-0.034X3-0.140X4-1.114X5+2.982X6. The accuracy was 76.5%(88/115) in retrospective assessment and 77.8%(21/27) in prospective assessment.

CONCLUSIONThe combined use of MAS and CT imaging signs is useful in identifying the pathological types of acute appendicitis in adults, so it is helpful in choosing reasonable therapeutic option for surgeons.

Acute Disease ; Appendicitis ; diagnosis ; Case-Control Studies ; Humans ; Retrospective Studies ; Sensitivity and Specificity ; Tomography, X-Ray Computed

To develop a stable cell line that could express the RSV NS1, the full-length RSV NS1 gene was generated by RT-PCR amplification from respiratory syncytial virus. NS1 gene was ligated with pBABE-puro to construct the recombinant retroviral expression plasmid pBABE-NS1, which was cotransfected into 293FT packaging cells with PIK packaging plasmid by calcium phosphate co-precipitation. The supernatant of 293FT was collected to infect HEp-2 cells, the resulting cell clones stably expressing NS1 were screened by puromycin. Using QPCR, CPE staining method and indirect immunofluorescence assay, the expression of NS1 at both gene and protein levels was identified. The recombinant plasmid pBABE-NS1 was identified by EcoRI and BamHI endonuclease digestion and the sequence analysis. QPCR results showed that the NS1 gene amplification in HEp-2-NS1 cells was 8483 fold higher than that in HEp-2 cells. Although the exogenous interferon was added, all cells were destroyed after 48 hours post infection using CPE staining method, showing that HEp-2-NS1 cells remained sensitive to the VSV virus. The results of RT-PCR and indirect immunofluorescence assay showed that the NS1 gene in HEp-2 cells could not only transcribe mRNA, but also express NS1 protein steadily. We had successfully established HEp-2-NS1 cell lines with stable expression of respiratory syncytial virus non-structural protein NS1.
Cell Line, Transformed ; HEK293 Cells ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; Respiratory Syncytial Viruses ; genetics ; Viral Nonstructural Proteins ; biosynthesis ; genetics

OBJECTIVETo explore the influence of obesity on surgical procedure and short-term surgical outcome in patients with gastric carcinoma.

METHODSA total of 426 patients with gastric carcinoma underwent laparotomy in our hospital during January 2006 and June 2008. All the patients were divided into obesity group and non-obesity group according to body mass index (BMI). The thickness of subcutaneous fat (SCF), abdominal anterior-posterior diameter (APD) and transverse diameter (TD) at the umbilicus level were measured by abdominal CT. Furthermore, the surgical data and postoperative conditions including short-term outcome were reviewed and compared between two groups.

RESULTSThe incidence of obesity was 29.8% in gastric carcinoma patients. Mean values of SCF thickness, APD and TD in obesity group and non-obesity group were (21.8+/-7.1) mm vs (14.4+/-7.5) mm, (223.2+/-24.6) mm vs (181.8+/-23.5) mm and (323.6+/-23.8) mm vs (285.8+/-24.4) mm (P=0.000). Longer operative time (P=0.007) and less amount of dissected lymph nodes were found in obesity group as compared to non-obesity group (P=0.000). Also, obesity group lasted a longer postoperative period of fever (P=0.000) and experienced more post-operative complications (P=0.005) than non-obesity group did.

CONCLUSIONSAbdominal CT scan may display the abdominal shape of gastric carcinoma patients, hence, it is useful to evaluate the difficulty of surgical procedure. These patients may involve in complicated surgical procedure and worse short-term outcome due to obese abdominal shape. Therefore, perioperative management should be emphasized for these patients.

Abdomen ; surgery ; Adult ; Aged ; Aged, 80 and over ; Body Mass Index ; Female ; Gastroplasty ; Humans ; Male ; Middle Aged ; Obesity ; Stomach Neoplasms ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome