Objective:To evaluate the identification rate of separating gel or HB&L pretreatment methods of MALDI-TOF-MS, thereby to provide a new idea for the rapid and accurate identification of pathogens of bloodstream infections in daily clinic practice.Methods:A total of 149 alarmed positive blood culture samples of single bacterial infection by routine laboratory methods were collected between January to December 2020 from the Sixth Medical Center, Chinese PLA General Hospital. Samples were pretreated with the separation gel accelerating tube method or the HB&L microbial culture system, followed by direct MALDI-TOF MS bacterial identification, the identification rates of the two pretreatment methods were compared and results from the traditional method were used as the standard control.Results:Among the 149 positive blood culture samples, 47.0% (70/149) were gram-negative (G -) bacteria and 53.0% (79/149) were gram-positive (G +) bacteria. Identification rate of G -strain level was 78.6% (55/70) by serum separation gel coagulation tube method and 91.4% (64/70) by HB&L microbial culture system, the difference was statistically significant ( P=0.033). Identification rate of G +strain levels was 73.4% (58/79) by serum separation gel coagulation tube method and 87.3% (69/79) by HB&L microbial culture system, the difference was statistically significant ( P=0.028). For G -bacteria in the range of 3.000-2.300, the identification rate was 22.9% (16/70) by serum separation gel accelerating tube method and 38.6% (27/70) by the HB&L microbial culture system, the difference was statistically significant ( P=0.044). For G +bacteria in the range of 3.000-2.300, the identification rate was 19.0% (15/79) by serum separation gel accelerating tube method and 34.2% (27/79) by the HB&L microbial culture system, the difference was statistically significant ( P=0.031). Conclusion:The identification rate of HB&L microbial culture system is higher than that of serum separation gel coagulation tube method. Direct MALDI-TOF MS identification of pathogenic bacteria in positive blood culture samples after pretreatment is feasible in daily clinical practice.

Objective:To investigate the effects of laparoscopy assisted complete mesocolon resection on the stress response and immune function of patients with colon cancer.Methods:From January 2013 to January 2016,130 patients with colon cancer in our hospital were selected and randomly divided into the observation group and the control group with 65 cases in each group according to the random lottery envelopes.The observation group was treated with laparoscopy assisted complete mesocolon resection,the control group was treated with open operation.The changes of semm IL-6,TNF-α and amours of CD4+ cells,blood loss,bowel function restoration time,hospital stays and complication rate of two groups were compared before and after treatment.Results:Both groups of patients were successfully completed the surgery and there was no serious complications occurred in both groups.The amount of bleeding,postoperative intestinal recovery time and postoperative hospital stay in the observation group were significantly less than those of the control group (P＜0.05).The chylous fistula,wound infection,pulmonary infection and complication rate on the 14th day postoperation in the observation group was 1.5％,which was 12.3％ in the control group and significantly higher than that of the control group (P＜0.05).The contents ofCD4+ cells in the observation group and the control group were 36.34± 7.83％ and 33.66± 9.82％,respectively,which were significantly higher than those before treatment(30.33± 6.49％ and 30.49± 5.33％)(P＜0.05),and the contents of CD4+ cells of observation group was significantly higher than that of the control group after treatment (P＜0.05).The serum levels ofIL-6 and TNF-α in both groups were significantly lower than those before treatment on the 14th day postoperation (P＜0.05),and the serum IL-6 and TNF-α levels of observation group were significantly lower than those in the control group on the 14th day postoperation (P＜0.05).Conclusion:Laparoscopy assisted complete mesocolon resection could promote the immune function and inhibit the stress response,which further promoted the rehabilitation and induced the complications in the treatment of patients with colon cancer.

Objective To evaluate the synergistic effect of everolimus on cisplatin-mediated cytotoxicity against human cutaneous squamous cell carcinoma COLO-16 cells.Methods Cultured COLO-16 cells were divided into several groups to be treated with everolimus at different concentrations of 50,100 and 200 nmol/L or 25 μmol/L cisplatin for 12 and 24 hours.Acridine orange (AO)-labeled autophagic vesicles combined with lysomal enzyme inhibitors (E64d and pepstatin) were used to detect the levels of autophagy and autophagic flow.Western blot analysis was performed to track the conversion of the autophagosome marker microtubule-associated protein 1 light chain-3 (LC3)-Ⅰ to LC3-Ⅱ,as well as to detect cleavage levels of Caspase 3 and poly-ADP-ribose polymerase (PARP).Lactate dehydrogenase (LDH) assay was conducted to detect cell death,and Annexin V-EGFP staining to evaluate cell apoptosis.Results The LC3-Ⅱ / LC3-Ⅰ ratios (LC3-Ⅰ conversion to LC3-Ⅱ) after 12-and 24-hour treatment did not differ among the 50-,100-and 200-nmol/L everolimus groups (12 hours:3.52 ± 0.21 vs.4.03 ± 0.39 vs.5.05 ± 0.22,P ＞ 0.05;24 hours:3.38 ± 0.26 vs.3.29 ± 0.06 vs.6.57 ± 0.16,P ＞ 0.05),but were significantly higher in the three everolimus groups than in the control group receiving no treatment (12 hours:2.07 ± 0.05,P ＜ 0.05;24 hours:2.61 ± 0.16,P ＜ 0.05).After 12-hour treatment,no significant differences were observed in the ratio of LC3-Ⅱ to β-actin between the 50-nmol/L everolimus + E64d + pepstatin group (1.26 ± 0.40),100-nmol/L everolimus ± E64d + pepstatin group (1.16 ± 0.34),200-nmol/L everolimus + E64d + pepstatin group (1.21 ± 0.39) and E64d + pepstatin group (1.19 ± 0.27,P ＞ 0.05).Moreover,there was no significant difference in the percentages of autophagic vesicle-positive cells between the 100-nmol/L everolimus + E64d + pepstatin group and E64d + pepstatin group (2.06％ ± 0.61％ vs.1.68％ ± 0.62％,P ＞ 0.05).After 24-hour treatment,the everolimus + cisplatin group showed significantly increased rate of cell death compared with the cisplatin alone group (42.58％ ± 0.93％ vs.18.20％ ± 1.46％).However,no significant differences were observed in the cleavage levels of Caspase 3 and PARP,the number of annexin V-labelled cells and ratio of LC3-Ⅱ to β-actin between the everolimus + cisplatin group and the cisplatin-alone group (P ＞ 0.05).Conclusion Everolimus has a synergistic effect on the cisplatin-mediated COLO-16 cell death,and this effect does not depend on cell apoptosis or autophagy.

Objective:To explore the prognostic value of poliovirus receptor(PVR) in patients with gastric cancer.Methods:Bioinformatics methods were used to analyze the expression of PVR mRNA in gastric cancer tissues and adjacent normal tissues and its relationship with prognosis. The clinicopathological data of 73 gastric cancer patients who underwent surgical treatment were retrospectively analyzed. Immunohistochemical method was used to detect the expression level of PVR in gastric cancer tissue and adjacent normal tissue, and its relationship with the clinicopathological characteristics of gastric cancer patients was analyzed.Results:The expression level of PVR mRNA in gastric cancer tissues was higher than that in adjacent normal tissues ( P<0.05). The expression of PVR is related to the clinical prognosis of gastric cancer patients, and the higher the expression of PVR, the lower the overall survival rate ( P=0.011) and the disease-free survival rate ( P=0.032) of gastric cancer patients. The results of immunohistochemical staining showed that PVR was highly expressed in gastric cancer tissue, mainly in the cytoplasm, while it was not expressed or lowly expressed in normal gastric tissue. High expression of PVR was associated with tumor size ( P=0.001) and recurrence 3-years after surgery ( P=0.013). Conclusions:PVR is overexpressed in gastric cancer tissue, and gastric cancer patients with high expression of PVR have a worse prognosis. The level of PVR has great value for the prognosis evaluation of gastric cancer patients.

Objective To explore the clinic application value of ultrasonography examination of optic nerve sheath diameter(ONSD) in brain injury.Methods From July 2008-June 2011,90 cases of brain injured patients were chosen as experimental group including light (A group),medium (B group),and heavy (C group) brain injured patients according to the admission GCS score ;50 cases of conventional physical examination and 90 cases of volunteers 50 in neurosurgical outpatient were chosen as control group.The ONSD of both groups were measured 3 mm behind the globe through orbital using color sonographic with different time after admission.3 times measurements were carried out for every optic nerve sheath.All client's ONSD mean and standard deviation were calculated.In 0.5 h after color dopplar ultrasound examination,lumbar vertebra puncturing measured intracranial pressure in different groups.Results After admission (1d,3 d,7 d,14 d),the ONSD of A group was (4.54 ±0.32)mm,(4.42 ±0.30)mm,(4.44 ±0.32) m,and (4.43 ± 0.25) mm,respectively; The ONSD of B groups was (4.48 ± 0.28) mm,(4.52 ± 0.24) mm,(4.46 ±0.28)mm,and (4.38 ±0.22)mm,respectively; The ONSD of C group was (5.67 ±0.35)mm,(6.36 ± 0.42) mm,(5.65 ± 0.23) mm,and (4.76 ± 0.35) mm,respectively.After admission (1 d,3 d,7 d,14 d),the intracranial pressure (IP) of A group was (82 ± 11) mmH2O,(79 ± 12) mmH2O,(90 ±15) mmH2O,and (86 ± 14) mmH2O,respectively; The IP of B group was (78 ± 15) mmH2O,(85 ± 10)mmH2O,(78 ± 16) mmH2O,(80 ± 11) mmH2O,The IP of C group was (225 ± 26) mmH2 O,(288 ± 23)mmH2O,(256 ± 23) mmH2O,(122 ± 18) mmH2O,respectively.Group D had the ONSD average of (4.58± 0.41)mm and IP of (88 ± 10)mmH2O after eyeball 3-mm place.No difference was found between A and B,A and D,or B and D (P＞0.05) ; A difference was found between A and C,B and C,or D and C (t =12.24～24.67,P＜0.01).Conclusions The ONSD and IP in light medium brain injured patients had no change.In patients with severe brain injury,IP changed with the time after injury,the ONSD increased with the IP,the ultrasonography examination of ONSD with the important value in the diagnosis and treatment can respond the IP increase,which is a non-invasion,convenient,fast,and feasible method for evaluation of cranial high pressure.

Objective:To investigate the expression of myocyte enhancement factor 2C (MEF2C) protein in gastric cancer, and to study its relationship with pathology and prognosis.Methods:Using bioinformatics methods, the expression of MEF2C in gastric cancer patients and its relationship with overall survival were analyzed in the TCGA database. Sixty-six patients with gastric cancer were selected as the research object. The expression of MEF2C in gastric cancer was analyzed by immunohistochemical staining of postoperative gastric cancer tissue samples. By analyzing the expression of MEF2C in gastric cancer, combined with pathological information and postoperative follow-up information, the effect of MEF2C on the progression of gastric cancer was studied.Results:The high expression of MEF2C in gastric cancer tissue was related to tumor size and tumor recurrence (all P<0.05), but not related to tumor grade and lymph node metastasis (all P>0.05). Conclusions:The high expression of MEF2C protein can be used as a predictor for poor prognosis of gastric cancer.

OBJECTIVETo explore the value of ¹⁸F-FDG PET-CT in evaluating bronchial mucosa involvement in patients with saroidosis.

METHODSA retrospective analysis was conducted among 6 sarcoidosis patients with and 14 patients without bronchial mucosa involvement to collect the data including the standard uptake value (SUVMax/Mean) of ¹⁸F-FDG, serum angiotensin converting enzyme (sACE), and proportion of lymphocytes and CD4⁺/CD8 ⁺ T lymphocyte ratio in bronchoalveolar lavage fluid (BALF).

RESULTSThe lung focal SUV(Max/Mean) was higher in patients with bronchial mucosa involvement than those without (7.04 ± 5.83/5.00 ± 4.69 vs 5.68 ± 3.66/3.82 ± 2.39), but such differences were not statistically significant (P=0.565/0.495). The SUV(Max/Mean) of the hilum of the lung and the mediastina lymph nodes were significantly higher in patients with bronchial mucosa involvement (13.28 ± 5.57/10.48 ± 4.43 vs 6.20 ± 1.77/4.52 ± 1.43, P=0.0003/0.0002; 13.84 ± 4.35/9.69 ± 2.74 vs 7.16 ± 2.52/5.28 ± 1.77, P=0.0004/0.0004). The level of sACE and CD4⁺/CD8 ⁺ T lymphocyte ratio in BALF were also significantly higher in patients with bronchial mucosa involvement (60.58 ± 16.3 vs 49.16 ± 13.3 IU/L, P=0.045; 7.30 ± 5.0 vs 2.90 ± 3.1, P=0.026). The proportion of lymphocytes in BALF was comparable between the patients with and without bronchial mucosa involvement (44.10 ± 10.3% vs 35.30 ± 12.5%, P=0.148).

CONCLUSIONSFor patients with saroidosis, ¹⁸F-FDG PET-CT is useful in evaluating bronchial mucosa involvement, which is one of the key features of active sarcoidosis.

Bronchi ; pathology ; Bronchoalveolar Lavage Fluid ; CD4-CD8 Ratio ; Fluorodeoxyglucose F18 ; Humans ; Lymph Nodes ; pathology ; Peptidyl-Dipeptidase A ; blood ; Positron-Emission Tomography ; Respiratory Mucosa ; pathology ; Retrospective Studies ; Sarcoidosis ; diagnosis

Objective To investigate molecular mechanisms underlying the synergistic damage to the human squamous cell carcinoma cell line COLO-16 by everolimus and cisplatin.Methods In the signaling pathway experiment,COLO-16 cells were divided into 4 groups:control group receiving no treatment,50,100 and 200 nmol/L everolimus groups treated with 50,100 and 200 nmol/L everolimus respectively.In the combined experiment,COLO-16 cells were divided into another 4 groups:control group,50 nmol/L everolimus group,25 mol/L cisplatin group,and 50 nmol/L everolimus + 25 mol/L cisplatin group.Western blot analysis was performed to analyze changes in mammalian target of rapamycin (mTOR) pathway,Akt pathway,DNA damage-related pathway and Csk homologous kinase (Chk) pathway.Results After the treatment with everolimus at different concentrations of 50,100 and 200 nmol/L for 12 and 24 hours,the phosphorylation levels of mTOR at ser2448 and ser2481 as well as Rictor at thr1 135 in COLO-16 cells were all decreased compared with the control group.However,there were no significant changes in the phosphorylation levels of downstream signals ULK1 at ser757,p70 S6 at thr389 and PKCα at thr638/64.The treatment with everolimus did not change the total protein level and phosphorylation of Akt.After the treatment with cisplatin for 12 and 24 hours,the phosphorylation levels of Rictor at thr1135 and Chk1 at ser345 were significantly increased,but the treatment with everolimus alone showed no such effects.After the combined treatment with everolimus and cisplatin for 12 and 24 hours,the upregulation of Chk1 and Rictor phosphorylation were significantly inhibited compared with the cisplatin alone group.Conclusions mTOR signaling is sensitive to everolimus in COLO-16 cells,but its targeted pathway is not regulated simultaneously to develop a cascade reaction.Everolimus may increase the cisplatin-induced death of COLO-16 cells by inhibiting the activation of Chk 1,but can not aggravate DNA damage induced by cisplatin.