To introduce the prophylaxis methods and dealing experiences of the common malfunctions of electronic gastroscopy. To summarize the prophylaxis methods and dealing experiences of the common malfunctions of electronic gastroscopy in recent years. To do everything carefully in the prophylaxis of the common malfunctions could ensure the high definition and resolution of the electronic gastroscopy. To manipulate properly with effective methods of preventing and dealing common malfunctions in use of electronic gastroscopy can not only lower the cost of repair but also prolong the service life of the machine.

Objective To evaluate the therapeutic effect of recombinant adeno-associated viral vector (rAAV) expressing human tissue kallikrein gene (rAAV-HK) on insulin resistance and renal complications in tyDe2 diabetic rats. Methods Male Wistar rats were injected low dose streptozotocin and fed with high fat and sucrose diets to form type 2 diabetic model. rAAV mediated HK gene (HK group) or LacZ gene (LacZ group) were introduced to the diabetic rats, and their systolic blood pressure, fasting blood glucose and insulin, serum creatinine, urine creatinine, urine osmolarity and urine microalbumin were measured. The homeostasis model assessment of insulin resistance (HOMA-IR), urinary albumin excretion rate (UAER) and creatinine clearance rate (Ccr) were calculated. The expression of PI3-kinase p11o catalytic subunit (p110) and Akt phosphorylation on Thr-308 were detected by Western blot. The morphology of kidney wag observed. Results Delivery of rAAVHK resulted in a reduction in blood pressure at 2 weeks and the hypotensive effect lasted for the duration of the study. The HOMA-IR was significantly lower in HK group than LacZ group (4.76±0.33 vs 8.36±0.48, P<0.01) at the end of the study, fasting insulin level was reduced [(8.19±2.45 vs 13.85±3.76)mIU/L. P<0.01], but there was no significant change in fasting blood glucose [(13.09±3.01 vs 13.58±2.88)mmol/L].The phosphorylation of p11o and Akt Thr-308 were significantly decreased in skeletal muscle and liver in LacZ group and were almost corrected by HK gene therapy. The UAER and Ccr were significantly lower and urinary osmolarity were higher in HK-treated rats compared with LaeZ rats. Histological assessment indicated that the renal complication was relieved by HK gene delivery. Conclusion The rAAV-mediated HK gene delivery efficiently attenuated insulin resistance partly through PI3K/Akt pathway and diabetic nephropathy in type 2diabetic rats.

Objetctive To summarize the associated complications with circulatory support device,and provide reference for chnical practice.Method A total of 8306 consecutive patients who underwent open heart surgery,in Department of Post-operation Intensive Care Unit of the Cardiac Surgery,Anzhen Hospital,Capital Medical University,was retrospectively studied from January 2005 to February 2007.And the clinical data of 246 patients including 63 female and 183 male patients with mean age 56.7±14.2 years supported with various circulatory support devices for perioperative cardiorespiratory function failure in ICU were analyzed.Left ventricular assist device (LVAD) was used in 3 patients by the cannulation of the left alritan and ascending aorta.The extracorporeal membrane oxygenation(ECMO) was established in 48 patients for postoperative cardiorespiratory function failure.The vencarterial bypass was established by cannulation of the right atrium in 41 patients and femoral artery and of venovenons in 2 patients,and of the right atrium and ascending aorta in 5 cases,lntra-aortic balloon pumping(I-ABP)was performed via the femoral artery either percutaneonsly by the Seldinger technique in 195 patients.The cardiac operations included coronary artery bypass grafting (n=170),coronary artery bypass grafting with romoldingof left ventricle (n = 22),coronary artery bypass grafting with valvular operation (n=10),valvular operation (n=27),heart transplantation(n=8),correction of congenital heart defects(n=6),aortic operations(n=2).The duration of circulatory support ranged from 4 to 451 hours.Correlative complications of 3 kinds of circulatory support device were compared and repair of ventricular septal perforation in the wake of acute myocardial infarction (n=1).Results Seventy-eight (31.7%) patients died.Seventy-one(28.9% ) patients devdoped various complications including infection(n=27),renal failure required renal rephcement therapy (n=27),re-exploration for bleeding(n=24),haemolysis(n=6),limb ischemia(n=15),neurological complications(n=6),oxygenator failure(n=7) Conchusions The improvement of management to reduce complications may result in improved outcomes of patients supported with circulatory support devices.

OBJECTIVE To investigate the risk factors and treatment of nosocomial pulmonary fungal infection after open heart surgery.METHODS A total of 11040 consecutive patients who underwent open heart surgery were retrospectively studied from Jan 2004 to Dec 2006.And the clinical data of 324 patients with microbiologically documented nosocomial infection were analyzed.RESULTS There were 61 infected cases caused by fungi,accounted for 18.8% of all infections during the same period.Sixteen patients died.All infected patients had serious original heart diseases,and received previously glucocorticoids and broad-spectrum antibiotic therapy.The number of senile and pediatric patients was 28.Forty three patients developed postcardiotomy cardiorespiratory and renal function failures,and circulatory assist was established in 22 patients.Thirty six patients prolonged mechanical ventilation time for over one weeks.Thirty four patients developed hyperglycosemia.Candida albicans was the predominant pathogen(39 strains,63.9%),followed by C.glabrata(15 strains,24.6%).Results of susceptibility test showed that fluconazole,itraconazole and flucytosine presented highly antimicrobial activity.CONCLUSIONS Developments of nosocomial pulmonary fungal infection is closely associated with the severity of preoperative underlying heart diseases,prolonged mechanical ventilation and longer period of broad-spectrum antibiotic therapy.

Human trophoblast cells were isolated and cultured in vitro in order to investigate possible pathogenesis of intrauterine infection caused by HCMV. Trophoblast cells were obtained by compound enzymes digestion and discontinuous percoll gradient. Cells and purity were identified by using immunocytochemistry assay with anti-CK7, Vim and beta-hCG antibodies. HCMV AD169 strain replication in isolated trophoblast cells and cell apoptosis were detected at different time points post infection (p.i.). The results showed that highly purified trophoblast cells were obtained. Specific virus replication was increased dramatically at the 24th h p.i., and then increased slowly during 48 h and 72 h. Apoptosis rate of trophoblast cells infected with HCMV was (34.68+/-3.14)% at 24th h p.i., while that in control group was (15.32+/-2.34)% (P<0.05). It was suggested that highly purified trophoblast cells can be isolated by the simplified cell purification method. HCMV can infect human trophoblast cells, and be quickly replicated, resulting in the accelerated apoptosis of human trophoblast cells during early time.

This study examined the impacts of intrauterine murine cytomegalovirus (MCMV) infection on the long-term learning and memory of offspring. Sexually matured male and female BALB/C mice without MCMV infection were identified by ELISA and then mated. Seventy pregnant mice were randomly divided into the virus group (n=40) and the control group (n=30), in which the pregnant mice were subjected to placenta inoculation of MCMV suspension (1 μL, 1×106 PFU) or the same amount of cell culture medium, respectively, at gestational age of 12.5 days. Some pregnant mice [virus group (n=20), control group (n=15)] were sacrificed by cervical dislocation at gestational age of 18.5 days, and the head circumference and brain weight of the mouse fetuses were measured, and the MCMV infection in their brain tissues was detected by PCR. The other pregnant mice [virus group (n=20), control group (n=15)] delivered naturally, and the learning and memory capability of the offspring at 70-day-old was analyzed by Morris water maze test. The results showed that 28.57% mouse fetuses in the virus group developed viral infection in the brain. Their head circumference and brain weight were significantly reduced as compared with those in the control group (P<0.01). The Morris water maze test revealed that the mouse offspring in the control group found the platform with straight-line trajectories after training. In contrast, the counterparts in the virus group intended to enter the central area, but looked for the platform with a circular trajectory. And the infected mice exhibited prolonged swimming distance and swimming latency (P<0.01). It was concluded that: (1) placenta inoculation of MCMV can cause fetal brain infection and intrauterine development retardation; (2) the offspring of MCMV placenta inoculation mice showed a long-term decline in learning and memory capability.

Objective To study the effect of Jingyebaidu granules on treating cytomegalovirus ( CMV) infection during mid-pregnancy. Methods The sexually mature guinea pigs with no CMV infection history served as the subjects. Put the male and female ones in the same cages. Then the female ones were randomly divided into three groups during mid-pregnancy. Model control group:15 guinea pigs which were inoculated 1 mL suspension of GPCMV intraperitoneally. Jingyebaidu Medicine group:15 guinea pigs which were treated with Jingyebaidu(3. 09 mL·kg-1 ) through stomach perfusion after inoculation for 14 days. Normal control group:15 normal mid-pregnant guinea pigs. Viremia rates were examined 7 days after infection. All animals were sacrificed 20 days after infection. The placenta infection rate, pup infection rate, still-born rate were examined. Results Compared with the normal controls, the still-born rate was increased in model control group(8. 33% vs 34. 55%, P<0. 05). In comparison to the model control group, the GPCMV maternal infection rate(86. 67% vs 33. 33%), placenta infection rate (91. 67% vs 61. 22%), pup infection rate(90. 91% vs 48. 28%), still-born rate(34. 55% vs 15. 52%) were significantly decreased in the Jinyebaidu group (all P<0. 05). Conclusion Jinyebaidu granules could reduce maternal infection,pup loss, and placenta infection caused by CMV inoculation during mid-pregnancy.

The present study examined von Willebrand factor (vWF) levels and ADAMTS13 activity in pregnant and severe preeclamptic women in order to shed light on the prothrombotic state in severe preeclampsia. Thirty healthy women of childbearing age, 22 second trimester pregnant women, 30 third trimester pregnant women and 10 severe preeclamptic patients were recruited in this study. ADAMTS13 activity was determined by the FRETS-vWF73 assay and vWF antigen (vWF:Ag) levels by an enzyme-linked immunosorbent assay. The results showed that there were statistically significant differences in plasma vWF antigen levels between the severe preeclamptic and third trimester pregnant women, between third and second trimester pregnant women (P<0.05). The third trimester pregnant women had significantly lower plasma ADAMTS13 activity than second trimester pregnant women (P<0.05). Nevertheless, no significant differences in plasma ADAMTS13 activity were found between severe preeclamptic patients and the third trimester pregnant women (P>0.05). In conclusion, plasma ADAMTS13 activity is normal in severe preeclampsia despite the increased vWF:Ag levels. Prothrombotic state is involved in the pathogenesis of severe preeclampsia, as a result of endothelial injury.

The effect of human cytomegalovirus (HCMV) on invasive capability of early pregnant extravillous cytotrophoblasts (EVTs) was investigated in vitro. Primary EVTs were obtained by complex phosphoesterasum digestion and gradient centrifugation from villous tissue aseptically taken from healthy pregnant women. Cytokeratin7 (CK7), vimentin (Vim) and c-erbB-2 were immunocytochemically detected to identify source of cells, and HCMVpp65 antigen was assayed to determine the infection state of primary EVTs by immunocytochemical staining. The EVTs were divided into two groups: control group and HCMV group, and the expression of c-erbB-2, matrix metalloproteinase-2 (MMP-2) and MMP-9 proteins was detected in two groups by immunocytochemistry and Western blotting. Enzymic activity changes of MMP-2 and MMP-9 were tested by gelatin zymography in primary EVTs infected with HCMV. The invasion of primary EVTs was detected by cell invasion assay in vitro after they were infected by HCMV. The cell source identification showed that the cells obtained were highly-pure primary EVTs, and primary EVTs could be infected by HCMV. Primary EVTs could express c-erbB-2, MMP-2 and MMP-9 proteins, and as compared with control group, the protein expression was decreased significantly in HCMV groups (P<0.05). Primary EVTs could secrete active MMP-2 and MMP-9 in vitro, and the activity of two MMPs was decreased significantly in HCMV groups (P<0.05). The in vitro cell invasion assay showed that the number of primary EVTs permeating Matrigel in HCMV group was decreased (P<0.05). We are led to conclude that HCMV can infect primary EVTs and inhibit their invasion capability, suggesting that the impaired EVT's invasion capability might be related to the abnormal expression of c-erbB-2, MMP-2 and MMP-9 proteins.

In the present study, through a functional strategy, a metagenome library of the marine microbes from the surface water of the South China Sea was screened for beta-glucosidase and six positive clones were obtained. One of these clones, pSB47B2, was subcloned and further analysed in sequence. The result showed that there was an open reading frame for a novel beta-glucosidase, which was nominated as bgl1B. Using pET22b(+) as vector and Escherichia coli BL21(DE3) as host, Bgl1B was overexpressed recombinantly with high yield obtained and substantial enzymatic activity detected. The recombinant protein (rBgllB) was purified by Ni-NTA affinity chromatography and further biochemically characterized. The results indicated that, with pNPG as substrate, the optimum pH and temperature for the hydrolytic activity of rBgl1B were about 6.5 and 40 degrees C respectively. Under the optimum conditions, rBgl1B hydrolyzed pNPG with an activity up to 39.7 U/mg, Km and Vmax being 0.288 mmol/L and 36.9 micromol/min respectively. In addition, rBgl1B could also hydrolyze cellobiose, with a Km of 0.173 mmol/L and a Vmax of 35 micromol/min. However, we did not detect evident hydrolytic activity of rBgl1B to lactose, maltose, sucrose, and CMC. The enzymatic activity of rBgl1B to pNPG was stimulated to certain degrees by low concentration of Ca2+ or Mn2+, whereas it exhibited significant tolerance against high Na+. Distinguished from most of the beta-glucosidases derived from fungi, which display the highest activities under acidic conditions, rBgl1B exhibited relatively higher activity and stability at pH between 7.0 and 9.0.
Amino Acid Sequence ; Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Metagenome ; genetics ; Metagenomics ; methods ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Seawater ; microbiology ; beta-Glucosidase ; biosynthesis ; genetics