Catheterization ; instrumentation ; methods ; Endoscopy ; Humans ; Paranasal Sinuses ; Sinusitis ; therapy

Objective To express human B7.2 extracellular domain with prokaryote expression system and to evaluate its biological activity in vitro. Methods PCR was used to amplify the extracellular region of human B7. 2which contained both the IgV and IgC domains. The recombinant PGEX-4T-3/hB7. 2 (IgV+C) was obtained by cloning the PCR product into a prokaryote expression plasmid PGEX-4T-3 and was transformed into the host strain of DH5-α. The fusion protein consisted of GST and hB7.2(IgV+C) was identified by SDS-PAGE and Western blotting.T cell activation was observed by exposing purified T lymphocytes to the fusion protein and [3H]-TdR incorporation with the presence of the first signal imitated by anti-CD3 antibody. Results The fusion protein GST-hB7.2 (IgV+C) was produced and detected in inclusive body form from engineered bacterial cells. With the first signal existed,T lymphocytes proliferated when it was co-stimulated by the fusion protein. Conclusion These results indicated that the functional human B7.2(IgV+C) fusion protein can be produced in bacterial cells and the fusion protein displays the co-stimulatory activity in T lymphocytes activation.

Objective: To analyze and evaluate the diagnostic value of plasma D-dimer combined with echocardiography and lower extremity vascular ultrasound for pulmonary thromboembolism (PTE). Methods: The clinical data of 140 clinically suspected PTE patients admitted to Department of Respiratory and Critical Care Medicine, Ruijin Hospital between January 2012 and December 2016 were retrospectively analyzed. Based on the results of computed tomographic pulmonary angiography (CTPA), all included patients were divided into CTPA negative group (non-PTE group) and CTPA positive group (PTE group). The univariate and multivariate analyses were used to compare and evaluate the diagnostic and predictive efficacy of plasma D-dimer combined with echocardiography and lower extremity vascular ultrasound for PTE. Results: There was no significant difference in gender, age, smoking, underlying diseases (malignancy, hypertension, coronary heart disease, dyslipidemia, and chronic obstructive pulmonary disease) and surgical history within 30 days between the two groups (All P>0.05). Univariate analysis revealed significant differences in D-dimer (P=0.003), pulmonary artery pressure (P=0.000), and partial pressure of carbon dioxide (P=0.000) between the groups. Multivariate analysis suggested that deep venous thrombosis of the lower extremities, pulmonary hypertension, and elevated D-dimer levels were significantly associated with the incidence of PTE. Further ROC curve analysis suggested that the area under the curve (AUC) of deep venous thrombosis, pulmonary hypertension and D-dimer increase was 0.624, 0.673 and 0.624, respectively. The AUC of combined three factors was 0.837. The sensitivity, specificity, positive predictive value and negative predictive value of the combined diagnosis reached 86.8%, 88.4%, 87.5%, and 84.6%, respectively. Conclusion: D-dimer combined with echocardiography and vascular ultrasound of lower extremity might have diagnostic and predictive value for PTE.

OBJECTIVETo study the effects of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion.

METHODCerebral ischemia model in gerbils was produced by ligating bilateral common carotid artery. The dynamic expressin of GFAP were determined by immunochemistry after cerebyal ischemia for 15 min followed by reperfusion for 24 hours and 48 hours.

RESULTGFAP positive reactions reached a peak after cerebral ischemia for 15 min followed by reperfusion for 24 hours. Bu Yang Huan Wu Decoction decreased the expression. GFAP positive reactions decreased after cerebral ischemia for 15 min followed by reperfusion for 48 hours, Bu Yang Huan Wu Decoction increased the expression.

CONCLUSIONThe regulation of Bu Yang Huan Wu Decoction on astrocytes after cerebral ischemia and reperfusion may be related to repairing process after cerebral ischemia.

Animals ; Astrocytes ; drug effects ; Brain Ischemia ; complications ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Hippocampus ; metabolism ; Male ; Plants, Medicinal ; chemistry ; Reperfusion Injury ; etiology ; metabolism ; pathology

Objective To observe the effect of electroacupuncture on early enteral nutrition (EEN) support in patients with severe craniocerebral injury.Methods A prospective, randomized, controlled study was conducted, 50 patients with severe craniocerebral injury admitted to the the Department of Intensive Care Unit of Traditional Chinese Medicine Hospital of Ningbo from January 2014 to October 2016 were enrolled, and they were randomly divided into electroacupuncture experimental group (26 cases) and conventional treatment control group (24 cases) by randome number table. Enteral nutrition (EN) support was implemented in 24 - 48 hours after admission for all the patients, additionally, the electroacupuncture experimental group was treated by electroacupuncture stimulating acupoint from the 1st day after admission, once 30 minutes daily for consecutive 10 days. The levels of EN calorie reaching standard situation and the parenteral nutrition (PN) necessary for addition were recorded on 3 days and 5 days after admission; the nasal feeding amount of the two groups was recorded on the 1, 4, 7 days; the incidences of complications during EN support period were observed in both groups.Results The proportion of EN calorie reaching the standard in electroacupuncture experimental group was significantly higher than that in the conventional treatment control group in 5 days of treatment [92.3% (24/26) vs. 70.8% (17/24),P < 0.05]. The proportions necessary for addition of PN support were lower in electroacupuncture experimental group than those in the conventional treatment control group on 3 days and 5 days of treatment [3 days: 19.2% (5/26) vs. 25.0% (6/24), 5 days: 7.7% (2/26) vs. 16.7% (4/24)], but there were no significant statistical differences between the two groups (bothP > 0.05). The nasal feeding amounts in electroacupuncture experimental group were significantly higher than those in the conventional treatment control group on 4 days and 7 days of treatment [4 days (mL): 1292.31±123.04 vs.1204.17±139.81,7 days (mL):1342.31±113.75 vs.1275.00±103.21, bothP < 0.05]. The incidence of complications of the electroacupuncture experimental group was significantly lower than that of the conventional treatment control group in 10 days of treatment [26.92% (7/26) vs. 41.67% (10/24),P < 0.05].Conclusions Application of electroacupuncture has a certain clinical value during implementing EN support for treatment of patients with severe craniocerebral injury, early intervention of electroacupuncture can effectively improve the patients' gastrointestinal function and elevate the successful rate of EN, and the therapy is simple, effective and safe without any obvious adverse reactions.

OBJECTIVETo investigate which of the two immunoglobulin (Ig)-like domains, the immunoglobulin variable region homologous domain IgV (hB7.2 IgV) and the immunoglobulin constant region homologous domain IgC (hB7.2 IgC) on the human B7.2 molecule contains receptor binding sites, and to evaluate whether the B7.2 protein expressed in bacteria has biological activity in vitro.

METHODSThree fragments of hB7.2 IgV,hB7.2 IgC and the complete extracellular region of human B7.2 containing both the IgV and IgC domains,hB7.2 Ig (V+C), were amplified by PCR and subcloned into pGEM-Teasy. Three recombinants,pGEX-4T-3-hB7.2 IgV,pGEX-4T-3-hB7.2 IgC and pGEX-4T-3-hB7.2 Ig (V+C), were generated by cloning the fragments into a prokaryote expression plasmid (pGEX-4T-3) and transformed into the host strain E. coli DH5alpha. The relevant target fusion proteins consisting of GST and hB7.2 IgV,hB7.2 IgC and hB7.2 Ig (V+C), were identified by SDS-PAGE and Western blotting. With the presence of the first signal imitated by anti-CD3 antibody, T cell activation was observed by exposing purified T lymphocytes to each soluble form of the three bacterially-produced human B7.2 fusion proteins by [(3)H]-TdR incorporation.

RESULTSThree recombinant fusion proteins of human B7.2, GST-hB7.2 IgV, GST-hB7.2 IgC and GST-hB7.2 Ig (V+C) were produced and detected in inclusion body form from engineered bacteria. With the first signal present,T lymphocytes proliferated when co-stimulated by bacterially-produced either GST-hB7.2 Ig (V+C) or GST-hB7.2 IgV fusion proteins, but not by GST-hB7.2 IgC.

CONCLUSIONSFunctional human B7.2 fusion protein can be produced in bacteria. The IgV-like domain of human B7.2 is sufficient for B7.2 to interact with its counter-receptors and co-stimulate T lymphocytes.

Antigens, CD ; pharmacology ; B7-2 Antigen ; Escherichia coli ; genetics ; Humans ; Immunoglobulin Constant Regions ; pharmacology ; Immunoglobulin Variable Region ; pharmacology ; Lymphocyte Activation ; Membrane Glycoproteins ; pharmacology ; Plasmids ; Recombinant Fusion Proteins ; pharmacology ; T-Lymphocytes ; immunology

OBJECTIVETo study mechanism of the apoptosis of rat pancreas islet β cell strain (INS-1 cells) induced by sodium arsenite.

METHODSINS-1 cells were exposed to sodium arsenite at the different concentrations. MTT assay was used to detect the viability of INS-1 cells. The potentials on mitochondrial membrane and lysosome membrane of INS-1 cells were determined with the fluorescence spectrophotometer. The apoptotic levels of INS-1 cells exposed to sodium arsenite were observed by a fluorescence microscope and flow cytometry.

RESULTSAfter exposure to sodium arsenite, the viability of INS-1 cells significantly decreased with the doses of sodium arsenite. At 24 h after exposure, the OD values of the mitochondrial membrane potentials declined observably with the doses of sodium arsenite (P < 0.01). At 48 h after exposure, the OD values of the lysosome membrane potentials significantly increased with the doses of sodium arsenite (P < 0.01). At 72 h after exposure, the apoptotic cells were observed under a fluorescence microscope and enhanced with the doses of sodium arsenite. The apoptosis cells with light blue, karyopyknosis, karyorrhexis, apoptotic body and chromatin concentration appeared. The results detected with flow cytometry indicated that after exposure, the apoptotic INS-1E cells significantly increased with the doses of sodium arsenite.

CONCLUSIONSThe sodium arsenite can induce the apoptosis of INS-1 cells through the mitochondria-lysosome pathway.

Animals ; Apoptosis ; drug effects ; Arsenites ; toxicity ; Cells, Cultured ; Insulin-Secreting Cells ; drug effects ; Lysosomes ; metabolism ; Membrane Potentials ; drug effects ; Mitochondria ; metabolism ; Rats ; Sodium Compounds ; toxicity

Objective To investigate the effects of nutritional therapy under energy metabolic monitoring on nutrition indicators and clinical prognosis of elderly patients with critical severe diseases in the department of Intensive Care Unit (ICU).Methods One hundred and twenty elderly patients admitted to the Department of ICU of Integrate Traditional Chinese Medicine Hospital of Ningbo, from January 2013 to December 2016 were enrolled, and they were divided into a control group (62 cases) and an observation group (58 cases) by randomized block method. The patients in observation group received nutritional support treatment under the guidance of energy metabolic monitoring, the amount of nitrogen needed was measured every day, and appropriate energy was provided according to the amount of nitrogen and the ratio of heat to nitrogen; the patients in the control group were given the nutritional support program according to experience. The clinical efficacy was evaluated after 7 days of treatment in the two groups, the differences in hemoglobin (Hb), serum albumin (Alb), prealbumin (PA), weaning success rate within 7 days, duration of mechanical ventilation, length of stay in ICU, the standard rate of enteral nutrition (EN) in 7 days, parenteral nutrition support rate, reaching EN target calorie time, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score on the 7th day after admission and at discharge, the incidence of complications such as abdominal distention, stress ulcer, ventilator associator pneumonia (VAP), heart failure during nutritional treatment and mortality were observed and compared after EN support between the two groups.Results Compared with control group, after treatment Hb (g/L: 136.5±2.5 vs. 90.4±2.3), Alb (g/L: 35.7±4.6 vs. 32.8±4.2), PA contents (g/L: 211.0±20.8 vs. 190.9±30.7), weaning success rate within 7 days [55.2% (32/58) vs. 33.9% (21/62)], the standard rate of EN in 7 days in the observation group were obviously higher [82.8% (48/58) vs. 51.61% (32/62) allP < 0.05], but duration of mechanical ventilation (days: 8.8±3.5 vs. 11.1±4.0), length of stay in ICU (days: 21.2±5.0 vs. 25.9±6.5), parenteral nutrition support rate [29.3% (17/58) vs. 51.6% (32/62)], reaching EN target calorie time (days: 4.4±2.1 vs. 6.2±2.9), APECHE Ⅱ score 7 days after admission (18.7±5.8 vs. 20.8±8.1), APACHEⅡscore at discharge (13.0±5.2 vs. 15.6±4.5) and the incidence of complications such as abdominal distension [10.3% (6/58) vs. 41.9% (26/62)], stress ulcer [3.4% (2/58) vs. 12.9%(8/62)], VAP [22.4% (13/58) vs. 25.8% (16/62)], heart failure [15.5% (9/58) vs. 24.2% (15/62)] etc, were all lower in observation group (allP < 0.05), and 2 weeks later the mortality was significantly lower in the observation group than that in the control group [13.79% (8/58) vs. 22.58% (14/62),P < 0.05].Conclusions Nitrogen required in elderly patients critically ill patients with early determination, the supply of nutrients to guide empirical method is more accurate compared to the nutritional therapy. Nutritional support under energy metabolism monitoring can shorten clinical course, improve nutritional indicators and help reduce the risk of complications and death.

Immune checkpoint therapy is a new mode of tumor therapy. At present, many kinds of PD-1/PD-L1 inhibitors have been approved for clinical application on various tumors, such as malignant melanoma, lung cancer, bladder cancer, lymphoma, etc. However, with the gradual promotion and application in clinical practice, various immune-related adverse reactions caused by PD-1/PD-L1 inhibitors have attracted extensive attention. Especially in recent years, a number of adverse reactions related to endocrine gland injury have been reported. This paper reviews the research status and corresponding clinical treatment methods of endocrine gland related adverse reactions caused by PD-1/PD-L1 inhibitors.

Objective To explore the cause,urgent management,further treatment,outcome and prevention of internal carotid artery (ICA) injury during transnasal endoscopic surgery.Methods Five ICA injury happened during transnasal endoscopic surgery of sphenoidal sinus and (or) sellar region,involving 1 case of traumatic optic neuropathy,1 case of sphenoidal cyst,1 case of fungal sphenoid sinusitis,1 case of sellar adenoid cystic carcinoma and 1 case of pituitary tumor.These five cases were from three different hospitals in 1990 -2009,and the clinical data were collected and retrospectively reviewed.Results Injury of ICA was related with improper anatomic localization and operative procedures.The locations of injury were in cavenous segement in 3 cases,lacerum segment in 1 case,and clinic segment in 1 case,respectively.The types of injury included 3 cases of bleeding of laceration,1 case of carotid cavernous fistula and 1 case of pseudoaneurysme.Salvage sphenoid sinus packing with vaseline strip and subsequent intravascular stent graft implantation (2 cases) and endovascular embolization (2 cases) effectively controlled bleeding.Four cases were successfully treated without craniocerebral or ocular complications,only 1 case died of massive blood loss.Among 4 survival cases,1 patient abandoned further therapy,the other 3 patients were curcd of primary disease by reoperation or subsequent treatment.Conclusion Preoperatively,reading carefully the imaging data,intraoperatively,identifying anatomical positions accurately,performing proper operation,and stopping bleeding decidedly, postoperatively, seeking interdisciplinary cooperation to repair vascular damages,all these procedures can effectively reduce the surgical risk of ICA injury.