Microarrays have been emerged as new research tools for high throughput parallel gene expression profiling, large scale gene discovery, mutation and polymorphism analysis in an automatic way. We present a review of development of DNA microarrays in sepsis

ObjectiveTo investigate the association of bacterial translocation (BT) with cachexia and its impact on the outcome of gastric cancer patients.MethodsSixty cachectic gastric cancer patients,50 age- and sex-matched non-cachectic gastric cancer patients,and 55 healthy controls were enrolled in this study between January 2008 and July 2009.Polymerase chain reaction was performed to detect bacterial DNA in the peripheral blood of cancer patients and healthy controls,Cytokine levels were tested by enzyme-linked immunosorbent assay.Flow cytometry was used to detect immune indicators.All the enrolled patients were followed up for two years,and the two-year survival rate was calculated.ResultsThe BT ratio was significantly higher in cachectic patients than in non-cachectic patients (25.0％ vs.8.0％,P =0.019) and healthy controls (25.0％ vs.0.0％,P =0.000).BT-positive cachectic patients had significantly higher levels of IL-1α,IL-6,TNF-α,and IFN-γ compared with BT-negative cachectic patients ( P =0.012,0.003,0.036,and 0.017,respectively ) and BT-positive non-cachectic patients ( P =0.011,0.034,0.000,and 0.022,respectively).The two-year survival rate in BT-positive cachectic patients was significantly lower than in BT-negative cachectic patients (P =0.023 ).The levels of CD3 +T,CD4+ T,natural killer cells,and CD4 + T/CD8 + T in gastric cancer patients were significantly lower than in healthy controls ( P =0.023,0.031,0.016,0.041,respectively),whereas CD8 + T level was significantly higher in gastric cancer patients than in healthy controls (P =0.038).ConclusionBT may contribute to the development of cancer cachexia and influence the long-term survival of locally advanced gastric cancer patients.

Objective: To explore the association of the serum levels of cytokine IL-10 with the occurrence of cachexia from patients with low-third gastric cancer. Methods: Radioimmunoassay was used to examine the serum levels of IL-10 in 150 patients with low-third gastric cancer and 135 healthy controls. Results: The serum levels of IL-10 were significantly higher in patients with low-third gastric cancer than controls(Z=-11.862, P<0.01). The serum levels of IL-10 were significantly higher in patients with low-third gastric cancer of clinical stageⅢ/Ⅳ than those with clinical stageⅠ/Ⅱ(Z=-10.028, P<0.01). The serum levels of IL-10 were significantly higher in patients with cachexia than those without(Z=-10.369, P<0.01). Logistic regression analysis showed that IL-10 was associated with odds ratios of 1.599 (95%CI:1.299-1.870, P<0.01) for cachexia. Conclusion: The serum levels of IL-10 are possibly associated with the occurrence of cachexia from patients with low-third gastric cancer.

Objective To establish a rapid,accurate,specific quantitative assay for detecting B.pertussis,and apply to clinical diagnosis.Methods According to the specific sequence of B.pertussis IS481 gene,the primers and the fluorescence probe were designed and synthesized.Then a fluorescence quantitative PCR for detecting B.pertussis was developed.The specificity,sensitivity and reproducibility of the method were evaluated.255 specimens including 225 nasopharyngeal swabs from suspected pertussis patients and 30 normal nasopharyngeal swabs were detected by fluorescence quantitative PCR.Results A rapid specific quantitative method for detecting B.pertussis was established.The standard curve of the method indicated that there was a good linear relationship between the CT value and the template concentration with the correlation coefficient being 0.998.The linear range of the system was from 102 to 108 copies/μl and the minimum was 102 copies.It had a high sensitivity and good specificity.The intra.and inter-assay coefficients of variation were 5.78%-16.7% and 8.25%-14.9% respectively.The fluorescence quantitative PCR identified 41 positive results for specimens from suspected pertussis patients and results of 30 normal specimens were all negative.Conclusions The method can quantitatively detect the B.pertussis rapidly with high sensitivity and specificity,it can be applied to clinical diagnosis.

Objective To evaluate wall histological abnormalities 2 to 3 cm to the end in high or intermediate anal atresia in order to identify features that explain postoperative bowel dysfunctions.Methods Sixty anal atresia patients treated in the Capital Pediatric Institution between January 2008 and December 2012 were recruited in our study.36 patients were resected the terminal anal segment (3 cm).Compared with those 24 cases who were not.Resected samples were fixed for HE and immuno-histochemical stainings.Clinical data including sacral ratio (SR),age at operation,gender,bowel function were evaluated.Results There was no significant difference in patients' SR value,gender and age at operation between resected group and control group.The median follow-up period was 4.5 years.The rates of voluntary bowel movement,soiling (grade 1,2,3) were similar in both groups,however,the rates of severe constipation in resection group was significantly lower that in control group (3 ％ vs.21％,P ＜ 0.05) In the bowel wall of distal 2 cm anrectal canal,the connective tissue was found to be irregular and abnormally represented.Muscle coat was abnormal in all cases,showing the dysplasia circular and longitudinal layers.The number of enteric nervous system was significant fewer in distal 2cm anrectal canal than that in distal 3 cm(1.6 ±0.9 vs.5.6 ±1.8,t=11.715,P＜0.01).Conclusions Resection of terminal 3 cm at least of the atresia anal canal benefits postoperative bowel defecation function.

In this study, the natural biological inducer, rat regenerating pancreatic extract (RPE), was used to induce human amniotic mesenchymal stem cells (hAMSCs) into insulin-secreting cells. We excised 60% of rat pancreas in order to stimulate pancreatic regeneration. RPE was extracted and used to induce hAMSCs at a final concentration of 20 microg/mL. The experiment methods used were as follows: morphological-identification, dithizone staining, immumofluorescence analysis, reverse transcription-PCR (RT-PCR) and insulin secretion stimulated by high glucose. The results show that the cell morphology of passge3 hAMSCs changed significantly after the induction of RPE, resulting in cluster shape after induction for 15 days. Dithizone staining showed that there were scarlet cell masses in RPE-treated culture. Immumofluorescence analysis indicated that induced cells were insulin-positive expression. RT-PCR showed the positive expression of human islet-related genes Pdx1 and insulin in the induced cells. The result of insulin secretion stimulated by high glucose indicated that insulin increasingly secreted and then kept stable with prolongation of high glucose stimulation. In conclusion, hAMSCs had the potential to differentiate into insulin-secreting cells induced by RPE in vitro.
Amnion ; cytology ; Animals ; Cell Differentiation ; physiology ; Cells, Cultured ; Humans ; Insulin-Secreting Cells ; cytology ; Mesenchymal Stromal Cells ; cytology ; Pancreas ; physiology ; surgery ; Pancreatic Extracts ; pharmacology ; Rats ; Regeneration

OBJECTIVETo investigate the relationship between the presence of the TNF2 allele and plasma concentrations of tumor necrosis factor-alpha (TNF alpha) and soluble TNF receptor (sTNF-R) with the development of acute severe pancreatitis (ASP) and severe sepsis.

METHODSGenomic DNA was prepared from peripheral blood leukocytes. The TNF1 and TNF2 biallelic polymorphisms were identified by analyzing NcoI-digested DNA fragments obtained from PCR products. Plasma levels of TNF alpha and sTNF-R were measured by EASIA.

RESULTSThe overall TNF2 allele frequency in ASP patients was comparable to that found in healthy volunteers (29.2% vs. 29.3%, P > 0.05). Severe sepsis occurred in 26 of 72 patients. Patients with severe sepsis showed a significantly higher prevalence of TNF2 than those without (46.2% vs. 19.6%, P < 0.05). Plasma TNF alpha, sTNF-R I, and sTNF-R II levels were (36 +/- 31) pg/ml, (5.4 +/- 3.5) ng/ml, and (11.2 +/- 7.8) ng/ml, respectively, in patients with severe sepsis, and (31 +/- 25) pg/ml, (4.6 +/- 3.8) ng/ml, and (8.8 +/- 6.6) ng/ml in non-severe sepsis subjects. Differences in TNF levels were not statistically significant between patients with ASP and control group (P > 0.05). Moreover, there was no correlation between TNF2 allele frequency and TNFalpha levels [(37 +/- 31) pg/ml vs. (31 +/- 25) pg/ml in TNF2 group and TNF1 group, respectively, P > 0.05].

CONCLUSIONSOur results suggest that there is no relationship between ASP and the TNF2 allele, but that the TNF2 allele is associated with a susceptibility to severe sepsis as a result of ASP.

Acute Disease ; Adult ; Female ; Humans ; Male ; Middle Aged ; Pancreatitis ; genetics ; Polymorphism, Genetic ; Receptors, Tumor Necrosis Factor ; blood ; Sepsis ; genetics ; Tumor Necrosis Factor-alpha ; analysis ; genetics

Objective To investigate the association of bacterial translocation (BT) with cachexia in colonic cancer patients.Methods From September 2015 to May 2017 the clinical data of 292 colon cancer patients were studied at Qingdao Municipal Hospital.The bacteria in peripheral blood and mesenteric lymph nodes were detected by bacterial culture,and the bacterial DNA in peripheral blood was detected by PCR technique to determine the occurrence of bacterial translocation.Intestinal epithelial T-cell subsets and NK cells were evaluated using flow cytometry.Western blot and immunofluorescence were used to check tight junction proteins Occludin,Claudins-2,Zonula occluden-2 in intestinal epithelium.Fluorescence in situ hybridization and immunohistochemistry were used to detect the translocated bacteria and endotoxin.Results Compared with noncachectic patients,cachectic patients had a significandy higher BT ratio (27.8％ vs.7.2％,x2 =20.871,P ＜ 0.001).BT in the intestinal mucus layer was associated with lower levels of T-cell subsets and NK cells in the intestinal epithelium in BT(+) patients (CD3 + T:36.69％ ±5.87％ vs.41.63％ ±5.03％,CD4+T:44.08％ ±5.12％ vs.49.58％ ±7.01％,CD8+T:65.68％ ±5.51％ vs.61.43％ ± 5.58％,CD4+ T/CD8+ T:0.71％ ± 0.21％ vs.0.91％ ±0.23％,NK:27.86％ ± 3.93％ vs.34.69％ ± 4.52％,all P ＜ 0.01).Endotoxin was detected within the small intestinal wall in BT(+) patients and claudin-2 expression increased (0.63 ± 0.13 vs.0.21-± 0.06,t =-2.936,P ＜ 0.01),whereas Occludin and Zonula occluden-2 expressions decreased (0.37 ± 0.13 vs.0.84±0.17,0.16±0.07 vs.0.58±0.19,t=2.151,2.111,bothP＜0.05).Conclusions BTmay contribute to the development of colon cancer cacheria,and tight junction could be the gateway of BT.