AIM:To establish a method for obtaining specific cells in solid tumor tissue by sorting of CD11b + myeloid cells in hepatic metastases from colorectal cancer.METHODS:Tumor tissues were prepared into single cell suspension by mechanical method combined with enzyme digestion,and then the CD11 b + myeloid cells were isolated by flow cytometry.The sorted cells were identified by immunocytochemistry.The viability and morphologiy of the sorted cells were evaluated by Giemsa and Typan blue staining.The cell purity was evaluated by flow cytometry.RESULTS:Sufficient numbers of CD11b+cells with high purity were isolated by sorting with flow cytometry from the single cell suspension prepared by mechanical and enzyme digestion.The purity of the cells was confirmed by statistical analysis (P ＜ 0.05).The positive rates of the cells before and after sorting were significantly different (P ＜0.01).The positive cells were verified by immunocytochemical method.Meanwhile,the sorted cells had complete morphology and good activity.CONCLUSION:The CD11b + myeloid cells in solid tumor tissue can be isolated by flow cytometry from the machine-enzyme digestion suspension with high purity,good activity and complete morphology.

Objective:To analyze the mean levels of skeletal muscle mass and strength in elderly patients with type 2 diabetes mellitus(T2DM), and to investigate the effects of chronic inflammatory factors and oxidative stress on them.Methods:A cross-sectional study was conducted on 120 patients with T2DM aged over 60 years and 126 elderly patients without diabetes(the control group). Skeletal muscle mass, strength and serum levels of chronic inflammatory factors interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and 8-hydroxy-2′-deoxyguanosine(8-OHdG)were determined, and their effects on skeletal muscle mass and strength in elderly patients with T2DM were analyzed.Results:Compared with the control group, grip strength decreased in elderly patients with T2DM(25.03±7.85)kg vs.(29.52±7.73)kg( P<0.01), and skeletal muscle mass decreased(21.36±5.46)kg vs.(22.01±5.22)kg with no significant difference( P>0.05). Serum levels of 8-OHdG were higher in elderly patients with T2DM than in the control group(3.08±0.26)ng/L vs.(2.59±0.16)ng/L( P<0.01). Correlation and regression analysis results showed that 8-OHdG was an influencing factor for muscle strength in elderly patients with T2DM( R2=0.457)and that height and weight could be influencing factors for skeletal muscle mass in elderly patients with T2DM( R2=0.822). Conclusions:Skeletal muscle mass and strength decline in elderly T2DM patients, probably as a result of increased levels of oxidative stress.These findings may serve as evidence for sarcopenia intervention in elderly T2DM patients.

Objective To investigate the risk factors of complication rate after colorectal cancer operation.Methods This study recruited a total of 254 non-emergency colorectal cancer patients admitted to our hospital between December 2005 and December 2006,and then evaluated the effects of life style,preoperative factors and intraoperative factors on postoperative complications.Results Single factor analysis showed that the postoperative complication rate was not significantly affected by gender,age,obesity,palliative/curative resection,anesthesia style as well as preoperative drinking or smoking history.Preoperative complications(P=0.001),tumor stage and operation time(P=0.025) affected the postoperative complicatin rate.Multi-factor logistic regression analysis showed that preoperative complications were the only risk factor of postoperative complications [P=0.001,OR=5.871,(95% CI 2.958-11.651)].Conclusion Old age as such does not represent a contraindication for surgical treatment.Preoperative complications,operation time and tumor stage significantly affect the postoperative complication rate.Preoperative complications are the strongest risk factor of all.Therefore,reasonable perioperative managements and shortening operation time are the key to reducing postoperative complications.

Objective:To explore the expressions of miRNAs related to accelerating senescence in serum of the patients with amnestic mild cognitive impairment (aMCI), and to clarify their effects in the pathogenesis of aMIC.Methods:The levels of miRNAs related to accelerating senescence (miR-132, miR-193b, miR-130b, miR-20a, miR-296, miR-329 and miR-206) were measured in the serum of the patients with aMCI (aMCI group,n=66) and healthy controls(control group,n=76) using quantitative real-time PCR(qRT-PCR).The genes targeted by the altered miRNAs were predicted by TargetScan 6.0.DAVID was used to analyze the function of miRNA target genes.The serum levels of brain derived neurotrophic factor (BDNF) and silent in formation regulator 1(SIRT1) were measured by enzyme-linked immunosorbent assay (ELISA) method.Results:The expression levels of miR-206 and miR-132 in serum of the patients in aMCI group were significantly higher than those in control group (P<0.05).BDNF and SIRT1 were both target genes of miR-206 and miR-132.The levels of BDNF (29.50 μg·L-1± 3.13 μg·L-1) and SIRT1 (1.86 μg·L-1± 0.25 μg·L-1) in serum of the patients in aMCI group were both obviously lower than those in control group (BDNF: 32.29 μg·L-1±3.66 μg·L-1;SIRT1: 2.10 μg·L-1± 0.29 μg·L-1, P<0.05).Conclusion:The expression levels of miR-206 and miR-132 in serum of the aMCI patients are significantly up-regulated.Both of them might be involved in the pathogenesis of aMCI through inhibiting the BDNF and SIRT1 expressions.

Objective To investigate the relationship between the coagulation system status and the pulmonary hemorrhage in children with severe hand,foot and mouth disease(HFMD)and approach the clinical significance of early detection of coagulation function. Methods By prospective case design method,89 cases with HFMD admitted to Department of Critical Care Medicine of Hebei Provincial Children Hospital from July 2010 to July 2012 were enrolled. The children were divided into severe group(46 cases)and critical group(43 cases)according to the severity of disease,and the children in critical group were subdivided into survivor group(26 cases)and non-survivor group (17 cases). Forty-four healthy children with the same age and in the same period were served as healthy control group. The blood of children was collected immediately after admission for determination of blood routine, prothrombin time(PT),thrombin time(TT),activated partial thrombin time(APTT),fibrinogen(Fg),and D-dimer (DD). Results There were no significant differences in PT,TT,APTT and Fg among severe group,critical group and health control group(all P>0.05). The blood platelets count(PLT)in severe group and critical group was significantly lower than that in health control group(×109/L:245±130,237±156 vs. 389±120),while the DD was significantly higher than that in healthy control group(mg/L:0.34±0.67,0.41±0.08 vs. 0.24±0.13),and the DD in critical group was obviously higher than that in severe group(all P<0.05). The mortality rate in critical group was 39.5%,and there were no significant differences in PT,APTT,Fg,TT and PLT between survivor group and non-survivor group(all P>0.05),but the DD in non-survivor group was significantly lower than that in survivor group(mg/L:0.60±0.09 vs. 0.12±0.09,P<0.05). Conclusions In children with severe or critical HFMD, the coagulation factor and blood platelet were in a state of mobilization,mild consumption state with the existence of fibrinolytic inhibition,but without systemic bleeding tendency,therefore it is in a compensatory stage of disseminated intravascular coagulation(DIC),not the mechanism of pulmonary hemorrhage. The monitor of DD has its clinical significance in evaluations of the disease situation and its prognosis.

Objective To investigate the effect of the laparotomy and laparoscopic surgery on the stress parameters and complication of patients with gastric cancer. Methods A total of 96 patients diagnosed as gastric cancer and treated by surgery in our hospital from January 2015 to January 2017 were divided into open operation group and laparoscopy group according to the operation method,48 cases in each group. Compared the operation time,bleeding volume,dissected lymph node number,postoperative hospitalization duration and anus exhausting time and complications in 6 months after surgery. The levels of WBC,CRP,TNF-α, IL-6 in serum before and after operation were detected by enzyme - linked immuno sorbent assay and compared. Results Compared with the open operation group, the bleeding volume,postoperative hospitalization duration and anus exhausting time of laparoscopy group were better with less dissected lymph node number and longer operation time, the differences were extremely significant(P ＜ 0. 01); the WBC,CRP,TNF-α, IL-6 levels of laparoscopy group at 1 day after the operation were lower than those of open operation group(P ＜ 0. 05). The incidence of complication of laparoscopy group was 22. 8％, which was less than 54. 7％ of control group, the difference was significant(P ＜ 0. 05). Conclusion Compared with the traditional open operation, laparoscopic radical gastrectomy can shorten the hospital stays and reduce the intraoperative blood loss, the stress response and complication rate after operation.

Objective To explore the relationship between immune and pathogenesis of amyotrophic lateral sclerosis (ALS) in the investigation of neurofilaments phosphorylation and ultrastructure features in spinal cord ventral horn motor neuron injury mediated by immune.Methods Using transmission electron microscope,we studied the uhrastructure features of abnormal accumulations of neurofilaments (NF) in motoneuron of the spinal cord ventral horn,and immunohistochemically investigated neurofilaments phosphorylation.Results Electron microscope found that there was abnormal accumulation of interwoven NFs in motor neuronal perikarya and proximal axons.Immunohistochemical study revealed that the SMI-32 immunoreactive positive neurons (12.00?1.05),compared with control (18.00?1.83),were reduced (P

Objective:To study the clinical characteristics and risk factors of intrauterine Ureaplasma urealyticum (UU) infection in very low birth weight preterm infants.Methods:From March 2019 to February 2022, very low birth weight preterm infants with gestational age 28~32 weeks admitted to our hospital were enrolled in this single-center retrospective study. According to the UU test results of respiratory tract samples obtained within 24 h after admission, the infants were assigned into the UU group (UU-PCR positive) and the non-UU group (UU-PCR negative). SPSS 26.0 statistical software was used to compare the clinical characteristics, laboratory indices, and complications between the two groups. Risk factors of UU infection were calculated.Results:A total of 327 preterm infants were included: 45 in the UU group and 282 in the non-UU group. No significant differences existed in gender, gestational age, birth weight and delivery pattern between the two groups ( P>0.05). Compared with the non-UU group, the UU group had significantly higher incidences of premature rupture of membranes (PROM) and chorioamnionitis, elevated white blood cell and platelet counts, procalcitonin and C-reactive protein levels, total duration of oxygen use and ventilation, bronchopulmonary dysplasia, necrotizing enterocolitis and metabolic osteopathy ( P<0.05). Multivariate logistic regression analysis showed that PROM ( OR=5.444, 95% CI 2.749-10.781, P<0.001) and chorioamnionitis ( OR=2.161, 95% CI 1.048-4.454, P=0.037) were independent risk factors for UU infection. Conclusions:PROM and chorioamnionitis are risk factors for UU infection in very low birth weight preterm infants. For high-risk premature infants, the UU test should be completed as soon as possible after birth.

Background & Objective: Radiotherapy and temozolomide are the standard therapy for newly diagnosed glioblastoma multiforme (GBM). However, it is unclear whether adding another agent to the commonly used radiotherapy-temozolomide (RT + TMZ) benefits newly diagnosed GBM patients. The present network meta-analysis aimed to assess the efficacy of combining other agents with RT + TMZ for GBM treatment. Methods: A comprehensive literature search was conducted on PubMed, EMBASE.com, Web of Science, and the Cochrane Central Register of Controlled Trials from inception to September 23, 2014, to include all randomized controlled trials of RT + TMZ-based therapy in GBM patients. Pairwise and network meta-analyses were performed to compare the therapeutic regimens. Results: Seventeen studies involving 4,148 patients were identified. The results of pairwise meta-analysis indicated no significant differences among most comparison groups, except for bevacizumab + RT + TMZ versus RT + TMZ for progression-free survival (hazard ratio [HR] = 0.71, 95% confidence interval [CI]: 0.59–0.86; P = 0.000) and RT + TMZ versus RT alone for overall survival (HR = 0.71, 95% CI: 0.58–0.88; P = 0.001). The results of network meta-analysis also showed no significant differences in most comparisons; however, adverse events were more common among patients receiving additional therapeutic agents other than RT + TMZ. The ranking probability analysis indicated that bevacizumab + RT + TMZ and nimustine + cisplatin + RT + TMZ were associated with the best progression-free and overall survival, but they also caused the most adverse events in GBM patients. RT + bevacizumab + irinotecan had the highest probability of being the best regimen for minimizing adverse events. Conclusions: The addition of other targeted agents, particularly bevacizumab and nimustine, to RT + TMZ could be slightly effective for the treatment of newly diagnosed GBM patients; however, adverse events remained common.
Glioblastoma

Objective To investigate the degrees of injury severity of sepsis models made by different kinds of Escherichia coli. Methods The 152 mice were randomly divided into control group, DH5α group, 44102 group, and 25922 group, with 38 rats in each group. DH5α group, 44102 group and 25922 group were intraperitoneally injected with 300 μL of Escherichia coli DH5α, 44102 and 25922 at the concentration of 1.0 × 109CFU/kg to prepare sepsis models of different kinds of Escherichia coli. Control group was injected intraperitoneally with the same amount of normal saline. (1) After 8 h, four mice were taken from each group for peripheral blood bacterial culture . (2) After 12 h, ten mice in each group were used for measuring serum levels of TNF-α and IL-6 by enzyme-linked immunosorbent assay (ELISA). (3) Western blot assay was used to determine the serum levels of high-mobility group protein (HMGB1) in four mice of each group. (4) Ten mice in each group were used to measure serum levels of alanine transaminase (ALT), aspartate aminotransferase (AST), creatinine (CR) and blood urea nitrogen (BUN) by automatic biochemical analyzer. (5) After liver, lung and kidney tissues were fixed with formaldehyde, hematoxylin-eosin (HE) staining was performed (n=10 for each group). Results In DH5α group, 44102 group and 25922 group, bacteria, inflammatory cytokines TNF-α, IL-6 and HMGB1 protein, liver and kidney indicators ALT, AST, CR and BUN showed a sequential increasing trend (P＜0.01). The severe degrees of alveolar structure damage, hepatic cell infiltration and renal glomerular atrophy were DH5α group, 44102 group and 25922 group in turn. There were no obvious damages of lung, liver or kidney tissues in control group. Conclusion Escherichia coli 25922 induces severe sepsis injury and can be used to study the animal models of the initial inflammatory phase of sepsis. Escherichia coli 44102 induces moderate damage of sepsis and can be used in animal models that do not require definitive sepsis staging experiments. Escherichia coli DH5α induces less damage of sepsis and can be used to explore immunosuppressive therapy of the animal model of sepsis.