OBJECTIVE To evaluate the prevention of antibiotic-associated diarrhea(AAD) in surgical critical illness patients and strengthen the usage of antibacterials to AAD in hospital.METHODS Using prediction and retrospection to analyze the factors of AAD.RESULTS The main factors for AAD are changing antibiotics frequently,the age of patient,complicated disease of patient.CONCLUSIONS Rational use and rigorous control of antibiotics are the keys to prevent AAD in hospital.

BACKGROUND: It has been confirmed that platelet aggregation defect presents in patients with hemorrhagic thrombopathy, and platelet membrane glycoproteins play a significant role in the process of platelet aggregation.OBJECTIVE: To observe the expression changes of platelet membrane glycoproteins between patients with hemorrhagic thrombopathy and healthy people.DESIGN: Case-control analysis.SETTING: Department of Integrated Traditional Chinese and Western Medicine, Union Hospital, Tongji Medical College,Huazhong University of Science and Technology.PARTICIPANTS :① Seventy-nine patients with hemorrhagic thrombopathy who received treatment from January 2001 to March 2003 in the Department of Integrated Traditional Chinese and Western Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology were enrolled in this study. The patients, including 31 male and48 femlae, averaged (35.76±14.14)years old. They all agreed to participate in this study and met with the diagnosis of hemorrhagic thrombopathy revised in 1996.Informed consents were obtained from all the patients and their relatives.The course of multiple bleeding symptoms was 1 to 14 years. And the bleeding sites were totally 167: extremity petechia was found in 64 cases, rhinorrhagia in 33 cases, hypermenorrhea (more than 150 mL in each cycle) in 29 cases, gingival bleeding in 28 cases, ocular fundus bleeding in 8 cases and conjunctival hemorrhage in 5 cases.②lnclusive criteria: All the cases presented clinical manifestation of multiple focal bleeding;The laboratory tests including platelet counts,bleeding time and coagulation profiles were examined with no marked abnormal changes;According to hemorrhage sites, the patients were examined by specialists of internal medicine,otorhinolaryngology, gynecology, stomatology and ophthalmology, and no specific diagnostic focuses of infection were found; They had normal blood pressure and heart rate,and those with thrombocytasthenia had been ruled out through ristocetin test. ③Another 34 healthy volunteer donors, 15 males and 19 females with an average age (30.12±7.14) years,were selected as normal control group.METHODS:① Detection of platelet aggregation ratio: Adenosine diphosphate (ADP, final concentration 1.90 μmol/L),arachidonic acid(AA, final concentration 0.37 μmol/L),and platelet activating factor(PAF, final concentration 150 nmol/L)were used as aggregation inductors, and Chronolog 430 type aggregometry was used to detect platelet maximum agglutination ratio.When maximum agglutination ratio was within 21％ and 40％, it was considered as aggregation defect, and when below 20％, it was considered as aggregation absence.② Test of platelet membrane glycoprotein: 3.6 mL cubital venous blood was drawn from the patients and mixed with 20 g/L disodium ethylenediamine tetraacetic acid(EDTA-NA2) for the purpose of anticoagulation at 1:9, and the same procedure was performed on the blood samples from the normal control group. The plasma was isolated and collected, after being centrifuged, the supernatant liquid was discarded, and paraform was added to fix the rest of the platelet solution. The platelet concentration was adjusted to 3×107 L-1. 100 μL regulated platelet solution was extracted, and fluorescein isothiocyanate (FITC)-marked CD42b (anti-GP Ⅰ b),CD41 (anti-GP Ⅱ b), CD61 (anti-GP Ⅲ a), CD42b/CD42a (anti-GP Ⅰ b/Ⅸ ), CD41/ CD61 (anti-GP Ⅱ b/Ⅲ a) and CD62p(anti-P-selectin)clonal antibodies 200 μL were added respectively and well mixed,and then cultured for 30 minutes at room temperature away from light. Finally phosphate buffer solution was added to the solution till the volume reached 1 mL,and FACS420 flow cytometer was used to analyze the samples, one sample for 10000 platelets, and the results were demonstrated by positive fluorescence percentage.MAIN OUTCOME MEASURES: ① Platelet aggregation ratio. ② Expression of platelet membrane glycoproteins.RESULTS:All the patients and the healthy subjects were involved in the analysis of results. ①All the cases were found to have platelet aggregation defect or absence which could be induced by single or more platelet aggregating agents. No abnormal platelet aggregations were found in the control group. ② The fluorescence intensities of GPIb/Ⅸ, GP Ⅱ b/Ⅲa complexes, GPIb, GPⅢa and P-selectin in patients with hemorrhagic thrombopathy were lower than those in the healthy subjects (t =2.50-5.57,all P ＜ 0.05). The fluorescence intensity of GP Ⅱb in patients with hemorrhagic thrombopathy was slightly higher than that in the healthy subjects, but no significant difference was found. (t =0.86, P ＞0.05).CONCLUSION: Abnormal expressions of platelet membrane glycoproteins may be partial pathogenesis of hemorrhagic thrombopathy.

Objective To analyze hematosis of modified homemade Buxue decoction discriminate on patients with hemorrhagic anaemia after unilateral total hip arthroplasty and prognosis.Methods88 patients with hemorrhagic anaemia who underwent unilateral total hip arthroplasty in our hospital from February 2015 to Janurary 2017 were selected, they were randomly divided into the observation group and the control group, 44 cases in each group.The control group received pure western medicine after unilateral total hip arthroplasty, the observation group added modified homemade Buxue decoction discriminate on the basis of the control group.Clinical therapeutic effect on anemia, hematosis indexes including hemoglobin (HB), red blood cell (RBC), erythrocrit (HCT) in the two groups were compared, occurrence of complications during postoperative treatment, hip scores were used to compared prognostic differences in the two groups.ResultsTotal effective rate of the observation group 95.45% was significantly higher than the control group (81.82%) (P<0.05);there was no significant difference in Hb, RBC and HCT level before the treatment in the two groups, Hb (130.23±13.09)g/L, RBC (4.16±0.71)×1012/L and HCT (0.42±0.03) in the observation group after the treatment were significantly higher than the control group (P<0.05);complication rate in the observation group 9.10% was significantly lower than the control group (25.00%) (P<0.05);there was no significant difference in hip joint function scores before the treatment in the two groups, hip joint function score in the observation group after the treatment (79.73±5.42)scores was significantly higher than the control group (P<0.01).ConclusionTreatment effect of modified homemade Buxue decoction discriminate on hemorrhagic anaemia after unilateral total hip arthroplasty is significant, can promote recovery of hematosis, hip joint and motor function effectively.

Objective Vaccination is a most effective method for the prevention of severe diseases caused by pandemic influenza and microRNA ( miRNA) mediated gene silencing has offered a novel approach to the construction of new vaccines.Our study aimed to construct a recombinant influenza A ( H1 N1 ) virus with the PB1 gene that carries the target fragment of miRNA Let-7b. Methods After comparing the sequence of the A/Nanjing/108/2009 H1N1 viral fragments with that of Let-7b, we selected PB1 as the optimal gene sequence, inserted the Let-7b binding target gene into PB1, ligated the modified fragments with pDP 2000, and named the recombinant plasmids pDP-mu-PB1 and pDP-sclb-PB1, respectively.We co-transfected the MDCK and 293T cells with the recombinant and other seven plasmids and injected the supernatant into the allantoic cavity of the chickenembryo for virus propagation, followed by detection of the virus by hemagglutination ( HA) assay and measurement of the viral titer by TCID50 .We amplified the viral cRNA by RT-PCR and identified the viruses by agarose gel electrophoresis and nucleotide sequence analysis. Results PB1 was the optimal sequence ( 83 bp -107bp) for the attenuation of viruses.The HA-titers of miRT-H1N1 and scbl-H1N1 were 1∶32 and 1∶64, and their viral loads were 4.68 ×105 and 7.94 ×104 TCID50/mL, respectively.Nucleotide sequence analysis showed the expected fragment in the rescued virus. Conclusion A recombinant strain vaccine was successfully constructed, which has laid the foundation for fur-ther assessment of virulence.

Objective To prepare Exosomes secreted by mouse hepatoma cell (H_(22)) and heat stressed Exosomes (HS-Exo) derived from heat stress-treated mouse hepatoma cell (H_(22)), in order to study the possible anti-tumor immune mechanism. Methods Exosomes and HS-Exo were purified by serial ultracentrifugation and sucrose density gradiant centrifugation, and were observed and identified by electron microscope. The components and production of the protein and the effects of the host immune response against hepatocellular carcinoma of HS-Exo were observed by using Exosomes as the control. Their immunological factors were detected by Western blot. Lymphocyte proliferation and specific cytotoxic activity of mouse splenic cells were determined by MTT. CD4~+ and CD8~+ lymphocytes infiltration in mouse tumor tissues immunized by both were analysed by immunohistochemical staining. Results HS-Exo was similar in morphology to the Exosomes, the important immune-ralated protein expressed in HS-Exo was increased (P<0.05). HS-Exo immunized mouse group showed more effective inhibition of tumor growth, better-induced lymphocyte proliferation,more significantly enhanced the cytotoxic activity of spleen lymphocytes, as well as a more prominent role in tumor therapy than Exosomes immunized mouse control group (P<0.05). Conclusions Heat stress treatment method for the preparation of HS-Exo was feasible. HS-Exo had a stronger role in the immuneactivity and tumor treatment than control Exosomes.

Objective To evaluate the clinical efficacy of the covered stent placements in the treatment of malignant esophageal stenosis and to analyze the interrelated factors and countermeasures of complications.Methods 102 patients with malignant esophageal stenosis were undergone treatment with the covered stent placements through the mouths under X-ray fluoroscopy.The stents included China-made and imports,and the specifications were various.92 patients underwent radiotheraphy before or after process.All cases were followed up after operations.Results The successful rate of operation was 100%,110 covered stents were placed in total.The clinical symptoms of patients disappeared or abated obviously.Complications included:chest pain in 36 cases(35.3 %),restenosis in 7 cases(6.9%),stomach-esophageal countercurrent in 6 cases(5.9%),stent migration in 6 cases(5.9%),esophageal bleeding in 4 cases(3.9%),esophagus-mediastinum fistula in 1 case(1%),stent jam in 1 case(1%)and stent fell off accompanied with rupture partially in 1 case(1%).The mean survival time was 10.6 months.Conclusion The covered stent placement in the treatment of malignant esophageal stenosis is a high effective and easy method,but it is not very safe.

AIM: In order to understand the pathological changes, characteristic of degeneration in optic nerve and retina after strike of optic nerve. METHODS: According to methods of Allen's spinal injury, a 600gcm-strike power was put on the intraocular portion of the optic nerve and created a striking injury on optic nerve. After a survival interval of 48 h, 1 week, 2 weeks, 4 weeks, 3 months, the animal's optic nerves and retinas were collected and fixed for morphological examination. RESULTS: Forty-eight hours after nerve injuries, the optic nerves were slight enlargement and vacuolation. In 1 week, the optic nerve began to degenerate in injured part and the glia cell had proliferated, but the forms of retinal ganglion cells(RGCs) were normal. In 2 weeks, the vacuolation and focal necrosis were appeared between nerve fiber. The number of RGCs began to decrease. Condensed nuclei presented in the retina. In three month, the diameter of the optic nerve decreased in injury part and collo-scar was formed. The phenomenon mentioned above was more obviously. The internal nuclear neurons and outer nuclear neurons appeared rare. The thickness of retina decreased. The number of RGCs began to decrease in 48 hours and progressed thereafter. It decreased about 3.35%, 13.23%, 19.74%, 23.20%, 29.28% in 48 h, 1 week, 2 weeks, 1 month, 3 months compared with the number of normal RGCs. RGCs began to apoptosis in 48 h. CONCLUSION: The model in this experiment could make definite uncompleted optic nerve and retina injuries. The degree of neuron injuries decreased from RGC, internal nuclear neurons to outer nuclear neurons. The number of RGCs began to decrease in 48 hours, and most quickly periods from 48 hours to one week.

Benign anastomotic stenosis after esophagogastrectomy could reduce the patients' quality of life,even resulting in severe malnutrition and death.The endoscopic treatment includes dilatation,stent insertion,locoregional injection,and a relatively new technique radial incision and cutting.This article reviewed the progress in endoscopic treatment of benign anastomotic stenosis after esophagogastrectomy.

Objective To elucidate the role of the Wnt/-catenin signaling pathway in regulating the phenotypic transformation of aortic valvular myofibroblasts to osteoblast-like cells.Methods Cultured primary valvular myofibroblastes isolated from porcine aortic valve leaflets were treated with oxidized low-density lipoprotein (ox-LDL) for different lengths of time:24 h,48 h and 72 h.The Wnt signaling pathway inhibitor Dickkopf-1 (DDK-1) was co-incubated with ox-LDL for 72 h.After cells harvest,the expression of myofibroblastic or osteoblast-like phenotype related markers,a-smooth muscle actin (α-SMA),bone morphogenetic protein 2 (BMP2),alkaline phosphatase (ALP) and corebinding factora-1 (Cbfα 1),was detected by Western blotting.The expression and sub cellular localization of β3-catenin was assessed by immunocytochemistry.Changes of the Wnt/β-catenin signaling pathway and the transformation of aortic valvular myofibroblast to osteoblast-like cells were monitored.Results BMP2,ALP and Cbfa 1 protein expression was not or barely detectable in the control group.However,after ox-LDL treatment,the expression of α SMA,BMP2,ALP and Cbfa 1 increased significantly (each P＜0.01) in a time-dependent manner (each P＜0.05).Besides,ox-LDL was also able to up-regulate the protein expression of β-catenin in a time-dependent manner (P＜0.05) and promoted its nuclear translocation.After DKK-1 treatment,the protein expression of β3 catenin and osteogenesis related markers was down regulated (P＜0.05).Conclusions The Wnt/β-catenin signaling pathway may play a crucial role in regulating the transformation of aortic valvular myofibroblasts to an osteoblast like phenotype.

Objective To evaluate the consistency and accuracy among 3 brands of flow cytometers (BriCyte E6,BD FACSCanto Ⅱ and Beckman Coulter FC 500) in the detection of lymphocyte subsets.Methods According to the methodology,the BriCyte E6 was compared with 2 flow cytometers commonly used in clinical detection.Seventy-three cases (40 male and 33 female) of anticoagulation peripheral blood specimens were collected in the clinical laborartory department of Xinhua Hospital in July 2015 and the percentage (％) and absolute number (#) of the lymphocyte subsets were detected by 3 different flow cytometers within samples collected 4 h.Results There were good consistency among the 3 flow cytometers (R2 ＞0.95,R2 from 0.969 5 to 0.992 4) in the detection of lymphocyte subsets percentage,so did in the detection of absolute number (R2 ＞ 0.95,R2 from 0.969 1 to 0.993 3).As to the precision evaluation,in the detectionof CD8％,T#,CD4+ T# and CD8+ T#,BriCyte E6 achieved a low CV％ compared with FACSCanto Ⅱ and FC 500 (Friedman statistics are 16.720,11.840,15.760 and 15.430,P =0.000 2,0.027,0.000 4,0.000 4,respectively).In the detection of T％,CD4％,NK％,B％,NK#,B#,there was no significant difference among the 3 flow cytometers (Friedman statistics are 4.242,3.916,0.852,2.595,1.835 and 0.578,P =0.119 9,0.141 2,0.653 2,0.273 3,0.399 6,0.749 0,respectively).Conclusions The 3 flow cytometers have a good consistency in the detection of lymphocyte subsets.BriCyte E6 may be an alternative or complement of existing flow cytometers.