[Establishment and validation of a predictive model for neurogenic urinary tract injury in children].

Objective: To establish a predictive model for upper urinary tract damage in children with neurogenic bladder and verify its efficacy. Methods: From January 2011 to December 2021, 143 children with NB in the Children's Hospital of Chongqing Medical University and 84 children with NB in the First Affiliated Hospital of Zhengzhou University were selected as the research objects. The former is set as the training set and the latter is set as the validation set, and the general parameters of the two are compared. The independent risk factors of upper urinary tract damage in children with NB were screened out by Lasso regression, and multivariate logistic regression analysis and a nomogram prediction model was established. The models were validated internally and externally on the training set and validation set, respectively, and the area under the receiver operating curve (ROC) was used to verify the accuracy of the model. Results: A total of 227 children with NB were included in this study, including 121 males and 106 females, aged (10.2±3.8) years. There was no significant difference in other parameters except age between the training set and validation set (all P>0.05); Lasso regression and multivariate logistic regression analysis showed that detrusor leakage point pressure (DLPP) ≥ 40 cmH2O (OR=4.76, 95%CI: 2.01-11.26, 1 cmH2O=0.098 kPa), overactive bladder (OAB) (OR=3.08, 95%CI: 1.34-7.04), bladder compliance (BC)<20 ml/cm H2O (OR=3.65, 95%CI: 1.41-9.47), history of previous urinary tract infection (OR=2.73, 95%CI: 1.09-6.81), and abdominal pressure/other voiding patterns (OR=2.86, 95%CI: 1.20-6.82) were risk factors for upper urinary tract damage in children with NB (all P<0.05). The above parameters were used to establish a nomogram model of upper urinary tract damage in children with NB. The internal and external validation results show that the AUC values for the training and validation sets were 0.84 (95%CI: 0.77-0.91) and 0.86 (95%CI: 0.79-0.94), respectively. Conclusion: The prediction model of upper urinary tract damage in children with NB constructed in this study has high discrimination, accuracy and clinical applicability, which can help clinicians identify high-risk patients and make individualized treatment design for these patients.

[Emulation pulmonary nodules localization model:a novel non-invasive localization technique in resection of pulmonary nodules].

Objective: To explore the accuracy and efficiency of a novel 3D-printed emulation localization model of small pulmonary nodules in lung surgery. Methods: From April 2020 to April 2021, a total of 66 patients were selected in the study, who underwent localization and resection of pulmonary nodules with video-assisted thoracoscopic surgery (VATS) guided by the 3D-printed emulation localization model at Department of Thoracic Surgery, West China Hospital of Sichuan University. There were 13 males and 53 females, aged from 25 to 79 (52.7±11.4) years. Of all patients, 24 (36.4%) had single pulmonary nodule, and 42 (63.6%) had synchronous multiple pulmonary nodules. The chest high-resolution CT image data were utilized for digital reconstruction and 3D printing to make a tailored life-size emulation pulmonary nodules localization model, which was used to navigate real-time intraoperative localization of nodules. Clinical data including operative parameters, localization information, resection types and pathological findings of nodules were analyzed. The pulmonary nodules that doctors planned to resect were categorized into two categories:major nodules and additional nodules, according to their presence of invasion and radiological risk factors. The accuracy of localization and resection efficiency of nodules were evaluated in accordance with the categories of the nodules respectively. Results: On the basis of preoperative evaluation, there were 71 major nodules with median maximal diameter of 0.9 (0.6-1.3) cm, and 77 additional nodules with median maximal diameter of 0.5 (0.4-0.7) cm. All patients underwent VATS surgery, 52 of them (78.8%) were treated with uniportal VATS and 14 (21.2%) with triportal VATS. Among the patients with single nodule, 18 segmentectomies and 6 wedge resections were performed; whereas among the patients with multiple nodules, 5 segmentectomies, 14 wedge resections, and 23 combined pulmonary resections (including 2 cases of lobectomy+segmentectomy, 7 cases of lobectomy+wedge resections, and 14 cases of segmentectomy+wedge resections) were achieved. The median operative time was 93 (45-240) min, and the median resection time for all nodules was 51.4 (6.7-147.0) min. All major nodules were successfully resected and visibly dissected after removal, and all additional nodules were successfully resected with 85.7%(66/77) nodules visibly dissected. The accuracy rate of localization of both types of nodules was 100%. All major nodules were malignant, and the malignancy rate of additional nodules was 21.2%(14/66). Conclusion: This novel 3D-printed emulation localization model of small pulmonary nodules proved to be a non-invasive, accurate and efficient technique. Not only that, it has a unique advantage in localization of synchronous multiple pulmonary nodules.

[A survey on the current development of thoracic surgery in tertiary hospitals of China].

Objective: To study the current development of thoracic surgery in China. Methods: Chinese Society for Thoracic and Cardiovascular Surgery and Chinese Association for Thoracic Surgeons jointly conducted a network survey to directors of thoracic surgery departments in the tertiary hospitals in China from November to December 2018. The contents of the survey included the basic information of the hospital and the status of thoracic surgery department in the hospital. Rank sum test was used to compare the data between different regional hospitals Results: A total of 636 tertiary hospitals participated in the survey. The total number of beds for thoracic surgery departments was 30 646, with M(Q(R)) of 40(20) (range: 3 to 393) for each hospital. The total number of thoracic surgeons was 6 747, with M(Q(R)) of 9(6) (range: 1 to 75) in each hospital. In 2015, a total of 312 425 operations were performed in the 636 hospitals, with M(Q(R)) of 268(484.5) (range: 4 to 8 320) for each hospital. The total number of lung cancer surgeries was 146 601 in 2015, with M(Q(R)) of 100(216) (range: 0 to 6 911) operations in each hospital. The total number of esophageal cancer operations was 67 076, with M(Q(R)) of 40(95) (range: 0 to 1 550) in each hospital. Minimal invasive thoracic surgery was performed in 94.3% (601/636) of the hospitals, with 86.6% (551/636) of hospitals carried out video-assisted thoracoscopic (VATS) lobectomy. Among the hospitals performing VATS lobectomy, 89.3% (492/551) of them started to perform the technique after 2006, and 93.1% (513/551) of them do single-direction thoracoscopic lobectomy. A total of 403 640 VATS lobectomies had been performed until 2015, including 163 682 cases of single-direction thoracoscopic lobectomy. In 2015, 73.74% (108 116/146 601) lung cancer operations and 37.44% (25 110/67 076) of esophageal cancer resections were performed by minimally invasive technique. The development level of hospitals among eastern, middle and Western China was different significantly on number of doctors, number of total operations, number of lung cancer surgeries, proportion of minimally invasive lung cancer surgery, number of esophageal cancer surgeries, and proportion of minimally invasive esophageal cancer surgery (χ(2)∶7.65 to 60.8, all P<0.05). Conclusions: The discipline of thoracic surgery, especially the minimally invasive thoracic surgery in China is now experiencing a rapid development. The proportion of minimally invasive lung cancer surgery is higher than that of in the developed countries. However, unbalanced development among different regions is still a great challenge in China.

[Strategies for the prevention and control of bleeding due to vascular injury in thoracoscopic lung surgery].

The technique of thoracoscopic lung surgery has gradually matured. Nowadays, thoracoscope is recommended as the most preferred approach for surgical treatment of early stage non-small cell lung cancer in different guidelines. However, there are still some cases of accidential major bleeding due to vascular injury during thoracoscopic lung surgery. The wall of the hilum vessels is relatively thin. These vessels often involve a great portion of the cardiac output blood flow. Once the injury happened, the emergent condition may be life-threatening due to massive blood loss. Therefore, this became an important factor which hindered the development of thoracoscopic lung surgery. In this review, details of the vascular injury in thoracoscopic lung surgery were summarized, including the incidence of vascular injury, commonly injured sites and reasons of the injuries. Among all the cases of thoracoscopic major pulmonary resection, 2.9% to 9.2% may suffer from vascular injury during the operation. The most commonly injuried sites are pulmonary artery and the branches, and this is also the most critical situation during thoracoscopic lung surgery. Hilum adhesion is the most important risk factor for vascular injury. On the one hand, the suction-compressing angiorrhaphy technique was developed for bleeding control and angioplasty. On the other hand, the strategies like pre-control of the pulmonary, cut the bronchus in advance, and fire the bronchus and pulmonary artery together may decrease the incidence of vascular injury in patients with risk factors.

[Clinical observation and related factors analysis of neonatal asphyxia complicated with retinal hemorrhage].

Objective: To observe and analyze related factors of neonatal asphyxia complicated with retinal hemorrhage. Methods: It was a retrospective case series. Seven hundred and twenty-one cases with neonatal asphyxia after 72 hours of birth were enrolled in this study. Fundus examination was performed on these newborns using the third generation wide-angle digital retina imaging system (RetCamⅢ), and the bleeding level was divided into level I, level Ⅱ and level Ⅲ. The conditions of the newborn and the mother during pregnancy were correlatively analyzed. The other factors were also analyzed including delivery mode, birth weight, gestational age, gender, grade of neonatal asphyxia, scalp hematoma, intracranial hemorrhage, fetal intrauterine distress, mother's age and antenatal complications. Single factor χ(2) test and multivariate logistic regression analysis were used to screen and judge risk factors causing retinal hemorrhage related to neonatal asphyxia. Results: In 721 cases of neonatal asphyxia, retinal hemorrhage was found in 204 newborns (28.29%). The hemorrhage was at level Ⅰ in 77 cases (37.75%) , at level Ⅱ in 38 cases (18.63%) and at level Ⅲ in 89 cases (43.63%) . Four cases also had vitreous hemorrhage. Asphyxia was mild in 673 infants (93.34%) and severe in 48 infants (6.66%). The difference in the degree of retinal hemorrhage between the patients with mild and severe asphyxia was significant (χ(2)=22.336, P=0.000). When asphyxia was aggravated, the degree of retinal hemorrhage increased. Relative factors analysis showed that delivery mode (χ(2)=158.643, P<0.05), gestational age (χ(2)=24.522, P<0.05), birth weight (χ(2)=11.916, P<0.05) and grade of neonatal asphyxia (χ(2)=19.809, P<0.05) had correlations with retinal hemorrhage. Logistic regression analysis indicated that grade of neonatal asphyxia and delivery mode were risk factors of retinal hemorrhage in neonatal asphyxia (OR=0.304, 0.085). Conclusion: The incidence of retinal hemorrhage in neonatal asphyxia was 28.29%. The degree of neonatal asphyxia and delivery mode may play roles in the occurrence of retinal hemorrhage in newborns with asphyxia. With aggravation of asphyxia, the degree of retinal hemorrhage may increase. (Chin J Ophthalmol, 2017, 53: 358-362).

HBV-induced ROS accumulation promotes hepatocarcinogenesis through Snail-mediated epigenetic silencing of SOCS3.

Interleukin-6 (IL-6) has been demonstrated to be involved in Hepatitis B virus (HBV)-associated hepatocarcinogenesis through activation of the STAT3 pathway. The sustained activation of the IL-6/STAT3 pathway is frequently associated with repression of SOCS3, which is both a target gene and a negative regulator of STAT3. However, the silencing mechanism of SOCS3 in hepatocellular carcinoma (HCC) remains to be elucidated. Here, we showed that the repression of SOCS3 and sustained activation of IL-6/STAT3 pathway in HBV-producing HCC cells were caused by HBV-induced mitochondrial ROS accumulation. Mechanistic studies revealed that ROS-mediated DNA methylation resulted in the silencing of SOCS3. Decreased SOCS3 expression significantly promoted the proliferation of HCC cells and growth of tumor xenografts in mice. Further studies revealed that HBV-induced ROS accumulation upregulated the expression of the transcription factor, Snail, which bound to the E-boxes of SOCS3 promoter and mediated the epigenetic silencing of SOCS3 in association with DNMT1 and HDAC1. In addition, we found that the expression of Snail and SOCS3 were inversely correlated in HBV-associated HCC patients, suggesting that SOCS3 and/or Snail could be used as prognostic markers in HCC pathogenesis. Taken together, our data show that HBV-induced mitochondrial ROS production represses SOCS3 expression through Snail-mediated epigenetic silencing, leading to the sustained activation of IL-6/STAT3 pathway and ultimately contributing to hepatocarcinogenesis.

Minimally invasive surgery for congenital diaphragmatic hernia: a meta-analysis.

OBJECTIVES: To compare the safety and efficacy of minimally invasive surgery (MIS) with traditional open surgical approach for congenital diaphragmatic hernia (CDH). METHODS: A literature search was performed using the PubMed database, Embase, and the Cochrane central register of controlled trials using a defined set of criteria. The outcomes, which include post-operative mortality, incidence of hernia recurrence, rates of patch use and complications, were analyzed. RESULTS: We investigated nine studies, which included 507 patients. All studies were non-randomized historical control trials. The MIS group had a significantly lower rate of post-operative death with a risk ratio of 0.26 [95% confidence interval (CI) 0.10-0.68; p = 0.006] but a greater incidence of hernia recurrence with a risk ratio of 3.42 (95% CI 1.98-5.88; p < 0.00001). Rates of prosthetic patch use were similar between the two groups. Fewer cases of surgical complications were found in the MIS group with a risk ratio of 0.66 (95% CI 0.47-0.94; p = 0.02). CONCLUSIONS: MIS for CDH repair is associated with lower post-operative mortality and morbidity compared with traditional open repair. Although rate of patch use appears to be comparable, the increased risk of CDH recurrence should not be ignored. The lack of well-controlled prospective trials still limits strong evaluations of the two surgical techniques.

Different sources of maternal selenium affect selenium retention, antioxidant status, and meat quality of 56-day-old offspring of broiler breeders.

The maternal effect of different Se source in offspring of young broiler breeders has been reported, but the lasting maternal effect of different sources of Se on offspring of breeders has received limited attention, so this study was conducted to investigate the effects of different maternal Se sources on Se retention, antioxidant status, and meat quality of 56-d-old offspring of broiler breeders. A total of two hundred forty 39-wk-old Lingnan Yellow broiler breeders were randomly distributed into 2 treatments, each of which was replicated 3 times with 40 birds per replicate, with a 14-d pretreatment and 56-d trial period. The treatments were fed a basal corn-soybean diet (0.04 mg∙kg⁻¹ Se) supplemented with 0.3 mg∙kg⁻¹ sodium selenite (SS) or selenomethionine (Se-Met). Fertile eggs were collected for incubation, after which 180 healthy chicks from each treatment were selected and randomly allocated into 3 replicates, with 60 birds per replicate. All the chicks were fed the same basal diet (0.04 mg∙kg⁻¹ Se) for 56 d. The Se concentrations in serum and tissues (liver, kidney, and breast muscle) of the 56-d-old offspring were significantly (P < 0.01) increased by maternal Se-Met intake compared with maternal SS intake. The antioxidant status of the 56-d-old offspring was greatly improved by maternal Se-Met supplementation in contrast with maternal SS supplementation, which was shown by increased glutathione peroxidase activity in serum and breast muscle (P < 0.01), glutathione concentration in serum (P < 0.05), and total antioxidant capability in pancreas (P < 0.01), as well as cytosolic glutathione peroxidase mRNA abundance in breast muscle, liver (P < 0.01), and pancreas (P < 0.05). The maternal Se-Met treatment was more effective in maintaining the shape of liver and pancreas cells, cell nuclei, chromatin, as well as cell membrane structure, and more organelles were observed in liver cells. The maternal Se-Met treatment had significant (P < 0.05) reduced the 48-h drip loss of 56-d-old offspring in comparison with maternal SS treatment. The results suggest that maternal Se-Met diet is superior to maternal SS diet in increasing Se retention and improving antioxidant status and meat quality of 56-d-old offspring.

Occipital somites guide motor axons of the accessory nerve in the avian embryo.

The accessory nerve (nervus accessorius) displays a unique organization in that its axons ascend along the rostrocaudal axis after exiting the cervical spinal cord and medulla oblongata and thereafter project ventrally into the periphery at the first somite level. Little is known about how this organization is achieved. We have investigated the role of somites in the guidance of motor axons of the accessory nerve using heterotopic transplantations of somites in avian embryos. The formation of not only accessory nerve but also the vagal nerve was affected, when a more caudal occipital somite (somites 2-4) was grafted to the position of the first occipital somite. Our study reveals that only the first occipital somite permits the development of ventral projection of accessory axons, a process that is inhibited by more caudal occipital somites.

The influence of different movements on ambulatory blood pressure in hypertensive subacute stroke patients.

OBJECTIVE: This study evaluated variation in blood pressure (BP) in hypertensive subacute stroke patients performing eight different types of active movement, and variations in BP over time. METHODS: The study included 35 subacute stroke patients (60 - 74 years old) and 15 age-matched healthy volunteers. Ambulatory systolic and diastolic BP was measured over 4 consecutive days, before and during active movement. RESULTS: The greatest effect of the different active movements in stroke patients was on mean systolic BP variability (BPV). There was a significant difference in systolic and diastolic BPV between stroke patients at different time-points and compared with healthy volunteers. Systolic BPV during shifting from the ward to the rehabilitation centre was significantly higher than for all other active movements. Mean systolic BPVs during the sessions on the first and second days were significantly higher than for the sessions on the third and fourth days in stroke patients and compared with healthy volunteers. CONCLUSIONS: Systolic BP was found to be increased in hypertensive subacute stroke patients during their first and/or second attempts at performing active movements. Therapists should consider the BP of hypertensive subacute stroke patients during these first two attempts, especially for activities involving the patient moving from the ward to the rehabilitation centre.

Simultaneous operation of concomitant diseases in the esophagus and lung.

The aim of this study was to retrospectively review our experience of performing simultaneous operations on concomitant diseases in the esophagus and lungs. From January 1998 to July 2009, simultaneous operations were performed on 13 patients with concomitant esophageal and pulmonary diseases, using coordinated surgical approaches. Among the 13 patients, six had primary cancers in the esophagus and lungs, five had primary esophageal cancer accompanied by a benign pulmonary disease, one had benign diseases in both esophagus and lung, and one had primary esophageal cancer with metastasis to the left lower lung. All patients survived the operations. Two major complications occurred postoperatively. One complication was bronchopleural fistula and the other was intrathoracic gastric laceration. Both patients recovered after additional treatments. Simultaneous operation of concomitant diseases in the esophagus and lungs is feasible and safe in selected patients who have received careful preoperative assessment, well-designed surgical approach, and proper perioperative management.

Deviation of type I and type II T cells and its negative effect on hematopoiesis in myelodysplastic syndrome.

Immunemediated hematopoietic suppression has been considered as one of significant pathophysiological changes in less-advanced myelodysplastic syndrome (MDS). To explore deviation of T cell subsets and its relationship to marrow cells apoptosis, measurement of helper-T (Th)/cytotoxic-T (Tc) subsets as well as the deviation situation within this two subsets (Th1/Th2 and Tc1/Tc2) in marrow was performed by flow cytometry from 39 MDS patients and 13 normal controls. Interferon (INF)-gamma and tumor necrosis factor (TNF)-alpha in marrow serum was simultaneously detected by ELISA (enzyme-linked immunosorbent assay). Furthermore, apoptosis rate of marrow cells was demonstrated by TUNEL (TdT-mediated dUTP nick end labeling). Results showed that Th and Tc subsets were unevenly activated, both deviating to type I response, which was especially obvious in patients with RCMD (according to WHO classification) and in lower-risk cases defined by International Prognosis Scoring System (IPSS). Level of INF-gamma/TNF-alpha in MDS marrow serum was markedly elevated, and so did the apoptosis rate of marrow cells. Although type I deviation was observed both in Th and Tc subsets, just Th1 cell percentage showed positive correlation with level of INF-gamma/TNF-alpha and apoptotic index of nucleated cells. In addition, cytokines level in marrow serum presented positive correlation to apoptosis. We then deduced that the increased Th1 cells in marrow may account for nucleated cells apoptosis in MDS through overproduced proapoptotic cytokines such as INF-gamma and TNF-alpha. Our results suggested that type I deviation of T cell subsets may play a role in pantocytopenia in MDS and the deviation pattern may be as a direct and effective parameter to predict response of immunosuppression therapy.

Skin biopsies demonstrate MPZ splicing abnormalities in Charcot-Marie-Tooth neuropathy 1B.

OBJECTIVE: To demonstrate that intronic mutations in the myelin protein zero (MPZ) cause Charcot-Marie-Tooth neuropathy 1B (CMT1B) by disrupting MPZ splicing. METHODS: We report a family with a T>G transversion at the invariant + 2 position in intron 4 of MPZ (c.614 + 2T>G) that abolishes 5' donor site recognition and is predicted to alter MPZ splicing. We obtained detailed clinical and neurophysiologic analysis of the family. We performed skin biopsies to investigate splicing abnormalities, MPZ protein levels, and localization in myelinated nerves. RESULTS: Patients developed a late onset neuropathy with minimally slow nerve conduction velocities. Skin biopsies confirmed the predicted skipping of exon 4 and downstream frameshift of the mutant MPZ. Quantitative immuno-EM demonstrated normal nerve MPZ levels, suggesting that the mutant MPZ was transported to compact myelin. CONCLUSIONS: Intronic mutations cause CMT1B by disrupting splicing and certain MPZ mutations may cause neuropathy by interacting with the wild type MPZ in the extracellular space of compact myelin.

Preparation of dendrimer-like polyamidoamine immobilized silica gel and its application to online preconcentration and separation palladium prior to FAAS determination.

A amino-terminated G 4.0 dendrimer-like polyamidoamine (PAMAM) immobilized silica gel (PAMAMSG) was prepared with a divergent method by repeating two processes: (1) Michael addition of methyl acrylate (MA) to surface amino groups; and (2) amidation of the resulting esters with ethylenediamine (EDA) from gamma-aiminopropyl silica gel (APSG) core. It was then used for the first time as microcolumn packing for the flame atomic absorption spectrometry (FAAS) determination of trace or ultra trace Pd(II), after flow injection (FI) online preconcentration and separation process. A limit of detection (LOD) of 3.9ngml(-1) was achieved when 0.200mugml(-1) Pd(II) was preconcentrated in 0.2moll(-1) HCl medium with a sampling flow rate of 6.0mlmin(-1) for 60s and the relative standard deviation (R.S.D.) was 1.7%. The proposed method was successfully used for the determination of Pd in two metallurgical samples.

Effect of ACE/NEP inhibition on cardiac and vascular collagen in stroke-prone spontaneously hypertensive rats.

Left ventricular remodeling in hypertension is associated with cardiac interstitial and perivascular collagen deposition. The dual angiotensin I converting enzyme/neutral endopeptidase inhibitor omapatrilat (also called vasopeptidase inhibitor) improves left ventricular remodeling in experimental heart failure. We hypothesized that omapatrilat would induce regression of cardiac and vascular fibrosis in hypertension. We, therefore, investigated the effect of omapatrilat on collagen deposition in heart and aorta of stroke-prone spontaneously hypertensive rats (SHRSP). Twenty-week-old normotensive Wistar-Kyoto (WKY) rats, untreated SHRSP, and SHRSP treated with omapatrilat (40 mg/kg per day, orally) for 10 weeks were investigated. Collagen in the heart and the descending thoracic aorta was stained with Sirius red. After 10 weeks, systolic blood pressure (BP) was significantly (P < .01) reduced in omapatrilat-treated versus untreated SHRSP. Interstitial collagen density was significantly decreased in the subendocardial myocardium (to 2.71 +/- 0.24% v 4.12 +/- 0.30%, respectively, P < .05) and in the midmyocardium of omapatrilat-treated versus untreated SHRSP (to 3.01 +/- 0.25 v 4.19 +/- 0.17% respectively, P < .05). Perivascular collagen was significantly (P < .05) decreased in the subepicardial, mid-myocardial and, subendocardial regions of the myocardium of omapatrilat-treated versus untreated SHRSP. Aortic collagen content decreased in omapatrilat-treated versus untreated SHRSP (to 36.1 +/- 2.8 v 58.8 +/- 6.1 x 10(3) microm2/mm section, respectively, P < .05). In conclusion, in addition to being a potent antihypertensive agent, omapatrilat significantly improves cardiac and vascular fibrosis in SHRSP.

Beneficial effect of glycoprotein IIb/IIIa inhibitor (AZ-1) on endothelium in Escherichia coli endotoxin-induced shock.

OBJECTIVE: To investigate the effects of AZ-1, a murine monoclonal antiglycoprotein-IIb/IIIa antibody, on endothelium and on hemostasis in a rabbit endotoxic shock model. DESIGN: Prospective laboratory study. SETTING: University laboratory. SUBJECTS: Thirty-five male New-Zealand rabbits. INTERVENTIONS: In vitro vascular reactivity, endothelium CD31-PECAM1 immunohistochemistry, plasma coagulation factors, and monocyte tissue factor determination were performed 1 day and/or 5 days after onset of endotoxic shock (0.5 mg/kg, intravenous bolus,Escherichia coli lipopolysaccharide) with or without treatment by AZ-1 (0.5 mg/kg intravenously) given 1 hr after lipopolysaccharide injection. MEASUREMENTS AND MAIN RESULTS: Metabolic acidosis and coagulation activation confirmed the presence of shock. AZ-1 treatment improved endothelial-dependent relaxation at 1 day (maximal effect = 87.2 +/- 4.0% vs. 60.9 +/- 5.2% in the nontreated group, p <.05) and at 5 days (maximal effect = 84.5 +/- 3.5% vs. 56.6 +/- 8.2% in the nontreated group, p <.05). Endotoxin-induced endothelial injury was decreased significantly by AZ-1 at 1 day (6.4 +/- 1.9% vs. 10.3 +/- 0.8% in the nontreated group, p <.05) and at 5 days (6.3 +/- 2.0% vs. 20.2 +/- 1.2% in the nontreated group, p <.05). Monocyte tissue factor expression was significantly reduced at 5 days. CONCLUSIONS: These data indicate that potent inhibition of platelet function via antiglycoprotein-IIb/IIIa receptor blockade can inhibit coagulation activation and protect against endothelial dysfunction and histologic injury in endotoxin-induced shock.

[Construction of human bone morphogenetic protein-2 expressing eukaryotic vector].

OBJECTIVE: To construct human bone morphogenetic protein-2(hBMP-2) expressing eukaryotic vector and observe whether it can be expressed in eukaryotic cells. METHODS: pUC19 was digested with Sal I and Xba I. The resulting Sal I-Xba I fragment (1.24 kb) which contains the full length of human BMP-2 cDNA was separated on agarose gel and ligated into eukaryotic expression vector pcDNA3 digested with XhoI and XbaI. The recombinant pcDNA3-hBMP-2 plasmid was transferred into fibroblasts cell line NIH3T3. The stable expression of hBMP-2 in the positive cells G418 selected was determined by in situ hybridization and immunohistochemical analysis. RESULTS: The two fragments digested from recombinant pcDNA3-hBMP-2 plasmid by EcoR I and Xba I represented 1.3 kb and 5.38 kb respectively by agarose electrophoresis, meanwhile the Xho I site was disappeared in pcDNA3-hBMP-2 indicating the successful construction of recombinant pcDNA3-hBMP-2 plasmid. Stable expression of hBMP-2 in pcDNA3-hBMP-2 transfected cells was confirmed by in situ hybridization and immunohistochemical analysis. CONCLUSION: hBMP-2 expressing eukaryotic vector is successfully constructed and can be expressed in eukaryonic cells.

Capillary electrophoresis system with flow injection sample introduction and chemiluminescence detection on a chip platform.

A capillary electrophoresis (CE) system with chemiluminescence (CL) detection was combined with flow injection (FI) sample introduction on a chip platform. A falling-drop interface was applied to perform FI split-flow sample introduction while achieving electrical isolation from the CE high voltage. A tubular reservoir at the capillary outlet served as both the CL reaction and detection cell for the luminol-peroxide-metallic ion chemiluminescent reaction, with the luminol included in the separation buffer and CL reagent H2O2 continuously introduced into the outlet reservoir. An optical fiber was positioned within the outlet reservoir directly opposite, and 300 microns away from, the capillary outlet for collecting and transferring the generated CL to the PMT. The peak height signals and the separation efficiency were almost independent of the reagent flow-rate, making the system a robust one. The performance of the system was illustrated by the separation of Co(II) and Cu(II), achieving baseline separation in 60 s. Detection limits (3 sigma) were 1.25 x 10(-8) and 2.3 x 10(-6) mol dm-3 for Co(II) and Cu(II), respectively. Peak height precision was 1.9% RSD (n = 9) at the 10(-7) mol dm-3 Co level.