Objective To compare the clinical effects of high flow nasal cannula (HFNC) and non-rebreathing oxygen face mask (NRB) in post-extubation patients.Methods 88 critically ill patients with machinery ventilations were divided into HFNC group and NRB group randomly.Blood gas analysis and hemodynamic parameters were assessed 1 hour prior to extubation and 6 hours after extubation.The primary clinical outcomes measured were ventilation-free days,re-intubation patient numbers,length of stay in ICU (Intensive Care Unite),total duration of hospitalization and mortality.The scant of breath degree and comfortableness of patient were recorded according to the Visual analogue scale.The measurement data were described by mean ± standard deviation ((x) ± s) and analyzed with t test,enumeration data were described by number of cases and composition ratio and analyzed with X2test,P ＜ 0.05 was considered to have statistical difference.Results There was no significant difference in clinical features between the two groups,The oxygenation index of HFNC group is significantly higher than that of NRB group after extubation [(251.4 ±43.9) vs.(201.7 ±60.7),P =0.037)].There were more ventilator-free days in the HFNC group than NRB group [(4.2 ± 2.1) vs.(3.4 ± 2.8),P =0.037)] and fewer patients required reintubation (P =0.028).The rate of ventilator associated pneumonia is also lower than NRB group (P =0.024).The patients' scant of breath feeling were obviously allevated comparing with the NRB group [(2.9 ± 1.1) vs.(3.7 ± 1.8),P =0.042)].The oxygenation index of NRB group significantly decreased after extubation [(242.9 ±68.4vs.201.7 ±60.7 P =0.048)].The two groups demonstrated similar hemodynamic patterns before and after extubation.And there were no statistically significant clinical differences in PaCO2,length of ICU stay,total duration of hospitalization or mortality.Conclusions Compared with NRB,HFNC is a more safe and effective clinical tool in the prevention and treatment of critical adult patients with extubation failure.

Objective To evaluate the effect of continuous blood purification (CBP) on cardiorenal syndrome (CRS) type Ⅰ.Methods Clinical data of 42 patients with CRS type [at our hospital were collected from January 2012 to June 2014.We observed and compared changes in mean arterial pressure (MAP),heart rate,respiration rate,acute physiology and chronic health evaluation (APACHE) Ⅱ score,and urinary volume before and 5 days after CBP.Meanwhile,levels of serum creatinine (Scr),cysteine proteinase inhibitor Cystatin C (CysC),serum creatinine (cTn) and B-type natriuretic peptid (BNP) were monitored.In addition,dynamic changes in cardiac index (CI),intrathoracic blood volume index (ITBI),global end-diastolic volume index (GEDI),central venous pressure (CVP),and extravascular lung water index (ELVWI) were monitored using the pulse induced contour cardic output plus monitoring system (PiCCO plus),and changes in left ventricular ejection fraction (LVEF) before and 5 days after CBP was measured by color Doppler ultrasound.Results There was no significant difference in MAP in patients with CRS type Ⅰ before and 5 days after CBP (P=0.08).Tacbycardia and tachypnea improved,while urine volume increased and the APACHE Ⅱ score decreased significantly,5 days after CBP(allP＜0.05).Plasma levels of Scr,CysC,cTn and BNP after treatment were lower than those before treatment [(126.8±68.3) μmol/L vs.(413.6±126.1) μmol/L,(1.1±0.8) g/L vs.(4.1±1.1) g/L,(2.6±0.4) μg/L vs.(3.5± 0.7) μg/L,(807.6±427.7) ng/L vs.(3300.3±567.6) ng/L,all P＜0.05)].Myocardial contractility,cardiac preload and lung related parameters also significantly improved after CBP (allP ＜0.05).Conclusions CBP can alleviate clinical symptoms of CRS type Ⅰ,improve cardiac and renal function,and is promising as an important auxiliary measure for the treatment of patients with cardiorenal syndrome type Ⅰ.

Objective To compare the therapeutic effects of continuous veno-venous hemofiltration (CVVH) versus repeated intermittent veno-venous hemofiltration (RIVVH) on patients with severe acute pancreatitis (SAP).Methods Fifty-six patients with SAP were randomly divided into the CVVH group (n =28) and the RIVVH group (n =28).The clinical symptoms and signs,the APACHE Ⅱ and MODS scores,the result of biochemistry including amylase and lipase,and the plasma levels of TNF-α,IL-6,IL8 before and after treatment,the duration of mechanical ventilation,boosting drug application time,the length of stay in ICU,the surgical intervention rate and the mortality were compared between the two groups.Results The clinical symptoms improved in the two groups after treatment (P ＜ 0.05).The APACHE Ⅱ and MODS scores were all reduced in the two groups after treatment (P ＜ 0.05).When compared with the RIVVH group,the result of biochemistry including amylase and lipase,and the plasma levels of TNF-α,IL6,IL-8 were significantly decreased (P ＜ 0.05).The duration of mechanical ventilation,the length of stay in ICU and the mortality were also significantly decreased in the CVVH group (P ＜ 0.05).Conclusions CVVH was more efficacious than RIVVH in the treatment of SAP.

OBJECTIVE To observe the curative effect of external dacryocystorhinostomy(EDCR) and intranasal endoscopic dacryocystorhinostomy(IEDCR)for chronic dacryocystitis, and to evaluatethe postoperative satisfaction of the patients.METHODS According to randomized controlled design, 108 chronic dacryocystitis patients were divided into the intranasal endoscopic dacryocystorhinostomy group and control group, both groups underwent intranasal dacryocystorhinostomy and external dacryocystorhinostomy respectively.The changes of symptom and clinical sign, operation time, hospital day, cost of hospitalization were recorded.All patients were followed up for 1-5years, and the postoperative satisfaction of the patients were investigated by interval satisfaction scale.RESULTS ①There were no significant difference between the groups at the curative effect, operation time and hospital day, the hospitalization cost of the intranasal endoscopic dacryocystorhinostomy group was higher than that of the control group;②The recurrence rate of the intranasal endoscopic dacryocystorhinostomy group and control group were 12.82% and 15.56% respectively after 5 years, Log Rank test ?2=0.394, P=0.530;③ The postoperative satisfaction of the patients in both groups were(81.70?10.242)and(75.72?10.653) score respectively, which were similar to normal distribution, t=-2.974, P=0.004.CONCLUSION The effect of intranasal endoscopic dacryocystorhinostomy for chronic dacryocystitis is similar to external dacryocystorhinostomy.But intranasal endoscopic dacryocystorhinostomy is a more ideal method, because it is minimal invasive technique and doesn't affect the face cosmetology.

To establish a gene regulation system compatible with biopharmaceutical industry and gene therapy, we constructed a fusion protein of biotin ligase from Bacillus subtilis (BS-BirA) and the trans-activation domain, and used its expression vector as the regulatory vector. Meanwhile, BS-BirA-specific operators were ligated upstream of attenuated CMV promoter to obtain the response vector. In this way, a novel eukaryotic gene regulation system responsive to biotin was established and named BS-Biotin-On system. BS-Biotin-On system was further investigated with the enhancing green fluorescent protein (EGFP) as the reporter gene. The results showed that our system was superior to the current similar regulation system in its higher induction ratio, and that the expression of interest gene could be tuned in a rapid and efficient manner by changing the biotin concentrations in the cultures, Our results show that the established system may provide a new alternative for the exogenous gene modulation.
Bacillus subtilis ; Biotin ; chemistry ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation ; Genetic Vectors ; Promoter Regions, Genetic ; Trans-Activators

Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
Animals ; CHO Cells ; Cricetulus ; Epitopes ; biosynthesis ; genetics ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; biosynthesis ; genetics ; Hepatitis B virus ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Objective To investigate the diagnostic value of ultrasonography for neck lymph node metastasis in papillary thyroid carcinoma(PTC) and Hashimoto's thyroiditis(HT) coexistent with PTC.Methods Two hundred and seventy-eight patients who accepted thyroid surgery were retrospectively reviewed for the pre-operative ultrasonographic and post-operative pathological reports.All patients were confirmed as PTC by surgery and pathology.According to the presence of HT confirmed in pathology,all patients were divided into two groups:group of PTC and group of HT with PTC.The status of neck lymph node metastasis and the diagnostic value of pre-operative ultrasound in detecting neck lymph node metastasis were studied.Results There were 185 cases in the group of PTC,and the rate of neck lymph node metastasis was 59.5 ％;while there were 93 cases in the group of HT with PTC,in which the rate of neck lymph node metastasis was 45.2％.The difference between the two groups in lymph node metastasis was statistically significant (P =0.024).The predictive accuracy of pre-operative ultrasound for central neck lymph node was 53.9％ in the group of PTC,which was statistically higher than 18.8％ in the group of HT with PTC(P =0.01).The predictive accuracy of pre-operative ultrasound for lateral neck lymph node was 79.4％ in the group of PTC,which had no statistical difference with that in the group of HT with PTC (73.1％,P =0.565).Conclusions The neck lymph node metastasis in PTC patients occurs more frequently than that in PTC patients with HT.The value of pre-operative ultrasound examination is lower for the detection of central lymph node metastasis,especially in PTC patients with HT;while ultrasound is more sensitive and accurate for lateral lymph node detection regardless of the existence of HT.

BACKGROUND: The increasing prevalence of diabetes mellitus has been become one of the diseases which threaten the heath of human being in the 21st century. Islet transplantation is considered to be the most effective approach to cure type Ⅰ diabetes mellitus. However, lack of donor tissue limits the application of this therapy. However, recent progress of stem cell research shows that stem cell therapy may be a potential means to solve this problem.OBJECTIVE: To take activin A and all-trans retinoic acid (AR) in inducing the differentiation of bone marrow mesenchymal stem cells (MSCs) and explore its possibility DESIGN: A randomized controlled experiment.SETTING: Institute of Biotechnology, Academy of Military Medical SciencesMATERIALS: This experiment was conducted at the Institute of Biotechnology, Academy of Military Medical Sciences from November 2004to June 2005. Six male Sprague-Dawley rats, with body mass of 150-160g, were provided by the Experimental Animal Center of Academy of Military Medical Sciences.METHODS: Femoral bone marrow of the rats was extracted under aseptic condition. Bone marrow mesenchymal stem cells (MSCs) were isolated with density gradient centrifugation. Passaged MSCs were randomly divided into 4 groups: high concentration of glucose (HG), AR, beta-mercaptoethanol (ME) and negative control groups. MSCs were induced to differentiate into IPCs with conditional medium containing high concentration glucose, activin A, RA and ME etc. After induction, phenotypes of differentiated cells were examined by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR).MAIN OUTCOME MEASURES: Expression of insulin and glucagon of differentiated cells were examined by immunocytochemistry. Insulin-1 mR-NA expression of differentiated cells was detected by RT-PCR.RESULTS: After bone marrow mesenchymal stem cells were induced,there were scattered insulin-and glucagon-positive cells in the HG group,many insulin-and glucagon-positive cells in the AR and ME groups, and these cells formed insulin-like structure. The expression of insulin-1mRNA could be observed in the HG, AR and ME groups. Insulin-and glucagonpositive cells and the expression of insulin-1mRNA were not observed in the negative control group.CONCLUSION: We adopt an induction scheme based on AR and other matured factors, and successfully make bone marrow mesench.ymal stem cells induce and differentiate into insulin positive reaction cells and form insulin-like structure, but its induction efficiency needs further improvement.

BACKGROUND:Islet beta cell replacement therapy is one of the most promising approaches for treating type 1 diabetes mellitus. However, its large scale application is hampered by a shortage of islet beta cells for transplantation. Pluripotent stem cells are one of ideal seed cells for islet beta cell replacement therapy, but pancreatic beta-cell differentiation is time-consuming and labor-intensive. OBJECTIVE:To construct a high efficient pancreatic and duodenal homeobox 1 (Pdx1)/insulin dual-reporter vector and to monitor the key genes expression during pancreatic beta-cell differentiation from pluripotent stem cells. METHODS:In order to construct a high efficient Pdx1/insulin dual-reporter vector, puromycin resistance gene was firstly introduced into pTiger vector, and then the original 410 bp mouse Ins1 promoter of the vector was replaced by 646 bp mouse Ins1 promoter. Finally, the dual-reporter vector was transduced into INS-1 and human induced pluripotent stem cells to testify its function. RESULTS AND CONCLUSION:The high efficient Pdx1/insulin dual-reporter vector was constructed successfully. The vector successfully acquired puromycin resistance gene and high gene expression efficacy of insulin in INS-1 cells. The specific gene expression pattern of Pdx1/insulin was first found in INS-1 cells. To conclude, the real-time monitoring function of Pdx1/insulin expression is preliminarily confirmed during pancreatic beta-cell differentiation.

With suspension adapted recombinant Chinese hamster ovary (CHO) cell lines 11G-S expressing human pro-urokinase (pro-UK) as the object of study, a serum-free medium for the cultivation of recombinant CHO cells in suspension was formulated by using Plackett-Burman design and response surface methodology. The two-level Plackett-Burman design was used to evaluate the effect of 10 medium supplements on the growth of the 11G-S cells in suspension culture. Among the 10 medium supplements, insulin, transferrin, and putrescine were identified as the most significant factors (P < 0.05). The response surface methodology with three factors and three levels was used to determine the optimal levels of these factors. And a serum-free medium, SFM-CHO-S for recombinant CHO cells suspension culture was formulated. The maximum cell density of 11G-S cells in SFM-CHO-S in suspension batch culture reached 4.12 x 10(6) cells/mL with a maximum pro-UK activity at 5614 IU/mL, which was superior to the commercial serum-free medium for recombinant CHO cells.
Animals ; CHO Cells ; Cell Culture Techniques ; methods ; Cricetinae ; Cricetulus ; Culture Media, Serum-Free ; Genetic Engineering ; Insulin ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; Transferrin ; pharmacology ; Urokinase-Type Plasminogen Activator ; biosynthesis ; genetics