Human ScFv against botulinum neurotoxin serotype A(BoNTa) was modified by fusing human IgG1 Fc to C terminal of ScFv. ScFv-Fc fusion protein was expressed at high level over 30% of total host cell proteins in E.coli. Recombinant protein existed in inclusion body form. Renatured ScFv-Fc was purified to 90%~95% by Protein G Sepharose column. In vitro ScFv-Fc could bind specific to toxiod BoNTa in ELISA. Recombinant ScFv-Fc had similar relative affinity to parent ScFv and had improved stability.

To explore the epidemiological character of Methicillin-resistant Stapkylococcus aureus (MRSA) by the phenotyping and genotyping motheds and to investgate the source, transmission, and the spread of nosocomial MRSA infection, consequently, reducing the nosocomial infection of MRSA. In this study, 19 MRSA strains were isolated from patients and environment in a hospital in two months. Patterns of resistantce against 16 antimicrobial agents and pulsed-field gel electrophoresis ( PFGE) of these strains were analyzed to find the relationship among those isolates Clustering analysis was made from the patterns. Some isolates with high homology was found in 19 MRSA, 11 of them belong to type A, and 8 of them belong to the same subtype A1. They were endemic in burn ward, oncological ward and ICU. In addition, 4 isolates were clustered into group B, all found in the same ward of burn unit Thus, our results indicated a outbreak of MRSA ( A type) in this hospital and the potential prevalence of MRSA (B type) , which might be mediated by health care stuff. It is essential to enhance the infection control implementation and to utilize the PFGE genotyping system for the real-time surveillance of MRSA.

Objective To assess the continent diversion results of sigma rectum pouch after radical cystectomy. Methods The reconstruction of bladder with sigmoid was modified for treatment of 18 cases of bladder tumor.The intestine was incised over a length of 20～24 cm with the junction of sigmoid colon and rectum as the midpoint so as to create a low pressure reservoir for urine and side-to-side anastomosis was performed on the posterior borders of the rectosigmoid wall.Submucosal tunnel modified technique was em- ployed in antireflux urethral implantation,Urination has been controlled by anal sphincter.Results About 80 minutes was spent to finish a new low pressure pouch after radical cystectomy.Among 18 patients with this op- eration,the controlled emiction were good after pull out the anal duct and"J"stent in 1 week to 2 months.Af- ter 2 months,the times of urination is stable,4～5 times in daytime and 1～3 times during nighttime.Two pa- tients had nocturnal enuresis and the symptom vanished after 2 months. One patient had adhesive ileus, two patients had hyperchloremia acidosis and kaliopenia,one patient had urethral stump cancer.There is no com- plication as anastomotic block,renal function lesion and severe upper urinary tract infection. Conclusion This operative method was easy,emiction control was well,and with higher quality of life for patients.It is al- so a better alternative diversion procedure that would be easily accepted.

Objective To study the effect of light sedation and traditional sedation (moderate sedation with daily sedation interruption) on hemodynamic indexes and prognosis in critically ill patients after cardiac surgery. Methods A total of 134 patients who were ventilated delay after heart surgery in our hospital from January to June 2017 were enrolled in this study. The patients were randomly divided into light sedation group (RASS score-1-1, n=65) and traditional sedation group (RASS score -3--2, n=69). All patients received sufentanil for postoperative analgesia. The light sedation group received propofol and/or dexmedetomidine as sedative drugs after operation, and the conventional sedation group used midazolam for postoperative sedation. The hemodynamic indexes, the first time of weaning off the ventilator, the duration of mechanical ventilation and ICU stay were compared between the two groups. Patients with low cardiac output syndrome after surgery were analyzed in subgroups. Results (1) There were no significant differences in heart function, operative complications and other indicators between the two groups after surgery (all P＞0.05). The low cardiac output syndrome was found in 12 patients in the light sedation group and 10 cases in the traditional sedation group. (2) Hemodynamic monitoring results displayed that the sedation/central venous oxygen saturation (SvO2/ScvO2) and cardiac index (CI) were higher after sedation than before sedation in both groups (all P＜0.05), but there was no significant difference between the two groups (all P＞0.05). Subgroup analysis showed that the SvO2/ScvO2index was higher in patients with low cardiac output syndrome in the traditional sedative group than that in the light sedation group (P＜0.05). There was no difference in the SvO2/ScvO2 index in patients with non-low cardiac output syndrome between two groups. (3) Compared with the traditional sedation group, the first off-line time, the total mechanical ventilation after surgery and the ICU stay time were significantly shortened, and the incidence of postoperative delirium was decreased in the light sedation group (all P＜0.05). Subgroup analysis showed that in patients with non-low cardiac output syndrome, the first off-line time, total postoperative mechanical ventilation time and total ICU stay were significantly shorter in the light sedation group than those in the traditional sedation group (all P＜0.05). There was no significant difference in patients with low cardiac output syndrome between the two groups (P＞0.05). Conclusion Patients with non-low cardiac output syndrome after cardiac surgery benefit significantly from the superficial sedative strategy, and the postoperative mechanical ventilation time and ICU residence time are reduced. The moderate sedation may contribute to the early cardiac function recovery in patients with low cardiac output syndrome.

OBJECTIVETo investigate the expression of human epidermal growth factor receptor 2 (HER-2/neu) in hepatocellular carcinoma (HCC) patients and its clinical significance.

METHODSThe expressions of HER-2/neu were detected by SP immunohistochemistry method in 30 patients with HCC, 10 with portal cirrhosis of the liver and 10 with normal liver.

RESULTSThe positivity rate of HER-2/neu was markedly higher in HCC patients than in those with portal cirrhosis and normal liver (Chi(2)=6.482, P=0.032). The expression of HER-2/neu was closely correlated to portal cirrhosis of the liver (P=0.041), tumor invasion (P=0.028) and Edmondson grades (P=0.012). The average survival time was significant shorter in patients with HER-2/neu-positive tumor than in those with HER-2/neu-negative tumor (P=0.036).

CONCLUSIONThe expression of HER-2/neu may play a role in the invasion, metastasis and progression of HCC. The patients positive for HER-2/neu in the HCC tissues have generally poor prognosis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Female ; Humans ; Liver Neoplasms ; genetics ; metabolism ; Male ; Middle Aged ; Neoplasm Metastasis ; Prognosis ; Receptor, ErbB-2 ; genetics ; metabolism

OBJECTIVETo investigate the effects of oxidized low-density lipoprotein receptor 1 (LOX-1) on secretion of adhesive molecules mediated by ox-LDL in human umbilical endothelial cells (HUVECs).

METHODSHUVECs with different concentration of ox-LDL (0, 10, 20, 50, 100 microg/ml) were incubated for 24 h, or HUVECs were pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for another 24 h. Expression of LOX-1 was determined by realtime RT-PCR and Western blot. mRNA and protein of ICAM-1, VCAM-1 and E-selectin were examined by RT-PCR and Western blot respectively.

RESULTSIncubation of HUVECs with ox-LDL (10-100 microg/ml) enhanced the expressions of LOX-1, ICAM-1 and E-selectin in a concentration-dependent manner (P < 0.01). On the contrary, ox-LDL did not affect the expression of VCAM-1 by HUVECs. The expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL were reduced in HUVECs pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for 24 h. This showed that both poly (I) and carrageenan obviously decreased the expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL.

CONCLUSIONox-LDL may upregulate the expression of LOX-1, ICAM-1 and E-selectin, and LOX-1 blocker may partly inhibit this upregulation. The results suggest that the expression of inflammatory molecules induced by ox-LDL in HUVECs is mediated by LOX-1.

Cell Adhesion ; Cell Adhesion Molecules ; Cells, Cultured ; E-Selectin ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; biosynthesis ; RNA, Messenger ; metabolism ; Receptors, Oxidized LDL ; metabolism ; Scavenger Receptors, Class E ; metabolism ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism

OBJECTIVETo investigate the value of the noninvasive examinations for localization of insulinoma.

METHODSThe clinical materials of 88 cases of insulinoma treated in Peking Union Medical College Hospital between January 2005 and November 2008 were analyzed retrospectively. There were 40 males and 48 females aged from 15 to 74 years old (averaged, 46.5 years old).

RESULTSThe positive rates for localization were 19.3% (17/88), 52.4% (11/21), 95.5% (64/67), 1/6, 30.0% (6/20), 83.9% (26/31), 8/8 and 5/5 respectively in transabdominal ultrasound, enhanced CT, multislice spiral CT pancreatic perfusion, MRI, somatostatin receptor scintigraphy, endoscopic ultrasound, laparoscopic ultrasound and intraoperative ultrasound. Thirty-one multiple tumors in 8 patients were resected. The positive rates of locating the multiple insulinomas of multislice spiral CT pancreatic perfusion and Intraoperative Ultrasound were 48.4% (15/31) and 100% (14/14) respectively.

CONCLUSIONSThe locating diagnosis of insulinoma has came to noninvasive examination era. Multislice spiral CT pancreatic perfusion is the first choice for localization. Intraoperative ultrasound might play an important role in locating tumors for patients with multiple insulinomas.

Adolescent ; Adult ; Aged ; Female ; Humans ; Insulinoma ; diagnosis ; Male ; Middle Aged ; Pancreatic Neoplasms ; diagnosis ; Retrospective Studies ; Young Adult

OBJECTIVETo determine the preoperative serum VEGF, IL-6, and CRP levels in colorectal carcinoma, and to explore their correlation with disease status and prognosis.

METHODSSerum VEGF and IL-6 levels were assessed using ELISA, and CRP was measured by immunoturbidimetry. They were compared between the colorectal carcinoma group and the control group. The five-year survival rate and poor prognostic factors were analyzed by Kaplan-Meier and Log-rank method, respectively.

RESULTSThe serum VEGF, IL-6, and CRP levels in colorectal carcinoma were (591 ± 312) pg/ml, (13.2 ± 3.7) pg/ml, and (1.14 ± 0.87) mg/dl, respectively, higher than that in the control group. The two groups showed significant difference in VEGF and CRP (P < 0.001, P = 0.002). VEGF expression was higher in male than that in female [(638 ± 387) pg/ml vs. (552 ± 271) pg/ml, P = 0.042]. The cases with tumor size smaller than 5 cm had lower VEGF expression compared with that in cases with tumor size ≥ 5 cm [(538 ± 275) pg/ml vs. (647 ± 331) pg/ml, P = 0.009]. IL-6 expression showed significant difference in males (11.7 ± 3.2) and females (15.2 ± 4.0) pg/ml, (P = 0.011). The five-year survival rate in the group with VEGF < 591 pg/ml was 86.8% (33/38), higher than that in the ≥ 591 pg/m group. High VEGF level tended to reduce survival (χ(2) = 0.933, P = 0.344). VEGF ≥ 591 pg/ml was a factor of poor prognosis in colorectal carcinoma, assessed by Log-rank methods (P < 0.05). Tumor size and VEGF concentration were risk factors of prognosis (P = 0.032, OR = 0.985; P = 0.011, OR = 0.976).

CONCLUSIONSSerum VEGF and IL-6 expressions have gender differences. Serum VEGF can be used as a biomaker of clinical diagnosis of colorectal cancer, and has an important significance on the prognosis of patients.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; C-Reactive Protein ; metabolism ; Colorectal Neoplasms ; blood ; pathology ; Female ; Follow-Up Studies ; Humans ; Interleukin-6 ; blood ; Logistic Models ; Male ; Middle Aged ; Preoperative Period ; Risk Factors ; Sex Factors ; Survival Rate ; Tumor Burden ; Vascular Endothelial Growth Factor A ; blood ; Young Adult

Diacylglycerol, DAG, because of its multifunctional and nutritional properties, attracted considerable attention recently. Enzymatic synthesis of diacylglycerols from linoleic acid was investigated in a solvent-free reaction in a continuously operated fixed bed reactors containing Lipozyme RM IM. By appropriate manipulation of the fluid-residence time, the relative proportions of the various acylglycerols in the effluent stream can be controlled. In addition, the presence of excess glycerol is effective for the removal of water produced during the esterification reactions. Under the conditions of molar ratio of linoleic acid to glycerol of 0.5, the immoblized enzyme maintained high stability and allowed the reaction to continue for 10 days without significant deterioration in enzyme activity. It was determined that the conversion of fatty acid, content of 1,3-DAG and volume efficiency of reactor reached optima under the conditions: a packaged-bed reactor(with a ratio of packed length to inner diameter of 7.8), reacting temperature at 65 degrees C, molar ratio of linoleic acid to glycerol of 0.5, and feeding flow rate of 1.2 mL/min.
Catalysis ; Diglycerides ; chemical synthesis ; Enzyme Stability ; Enzymes, Immobilized ; chemistry ; Lipase ; chemistry

Objective: To evaluate the prognostic value of kinetic changes in minimal residual disease (MRD) status, as well as its relationship with risk stratification, therapeutic response and treatment in patients with newly-diagnosed multiple myeloma (MM) . Methods: A total of 135 patients with newly-diagnosed MM were screened, and 105 patients who achieved VGPR or more as the best responses were included into this study. The MRD status was determined by multiparameter flow cytometry (MFC) at multiple intervals after two cycles of treatment until clinical relapse, death, or last follow-up. The statistical methods included Kaplan-Meier analysis, Cox regression, etc. Results: ①In all 135 patients, 57.8% (78/135) patients achieved MRD negativity (MRD(-)) after treatment. In 105 patients who achieved VGPR and thus included in this study, the MRD(-) rate was 72.4% (76/105) , with a median interval of 3 months from starting treatment to achievement of MRD(-) status. ②The 2-year PFS rate of patients with MRD(-) status was significantly higher than that of MRD(+) status (62.2% vs 41.3%, P=0.001) , while MRD persistence (MRD(+)) was an independent factor for poor prognosis (multivariate analysis for PFS: P=0.044, HR=3.039, 95%CI 1.029-8.974) . ③Loss of MRD(-) status (i.e., MRD reappearance) showed inferior outcomes compared with MRD sustained negative ones, the PFS was 18 months versus not reach (P<0.001) and the OS was not reach for both (P=0.002) . ④The 2-year PFS and OS rates of patients with duration of MRD(-)status≥12 months were significantly higher than those of the control group (PFS: 77.7% vs 36.7%, P<0.001; OS: 96.4% vs 57.9%, P<0.001 respectively) . Duration of MRD(-) status was associated with a marked reduction in risk of relapse or death (univariate analysis for PFS: P<0.001, HR=0.865, 95%CI 0.815-0.918; for OS: P=0.001, HR=0.850, 95%CI 0.741-0.915 respectively) . ⑤Moreover, even in patients carrying high-risk cytogenetic abnormalities (CA) or ineligible for ASCT, MRD negativity remained its prognostic value to predict PFS (high-risk CA medianPFS: not reach vs 19 months, P=0.006; ineligible for ASCT medianPFS: not reach vs 25 months, P=0.052 respectively) . ⑥Last, treatment with the bortezomib-based regimens contributed to prolonged MRD(-) duration (median MRD(-) duratio: 25 months vs 10 months, P=0.034) . Conclusion: Our findings supported MRD(+) status as an independent poor prognostic factor in MM patients, which implicated that duration of MRD(-) status also played a significant role in evaluation of prognosis, while loss of MRD(-)status might serve as an early biomarker for relapse. Therefore, monitoring of MRD kinetics might more precisely predict prognosis, as well as guide treatment decision, especially for when to start retreatment in relapsed patients.
Bortezomib/therapeutic use* ; Humans ; Multiple Myeloma/therapy* ; Neoplasm Recurrence, Local ; Neoplasm, Residual/diagnosis* ; Prognosis ; Risk Assessment ; Treatment Outcome