[The inhibitory effect of CtBP-interacting protein on the oxidative damage of cerebral endothelia cells].

Objective: To explore the effect of CtBP-Interacting protein (CtIP) on oxidative damage of cerebral endothelia cells and its mechanism. Method: Cerebral endothelia cells were stimulated by TBHP to induce oxidative damage. The cell line of CtIP gene were prepared by over-expression and interfering lentivirus technology. Cell damage was detected by immunofluorescence assay of cysteinyl aspartate specific proteinase-3 (Caspase-3). The expression of CtIP and Caspase-3 protein was detected by Western blotting, and the related genes of CtIP signaling pathway were detected by Realtime RT-PCR. Result: The results of immunofluorescence and Western blotting showed that overexpression of CtIP inhibited the Caspase-3 expression reducing to 1/3 level compared with normal cultured cerebral endothelia cells. Interfering with CtIP expression resulted in the Caspase-3 expression increased significantly to 4/5 level compared with normal cultured cerebral endothelia cells and cerebral endothelia cells were damaged more severely. Realtime RT-PCR data showed that expression of CtIP significantly increased the expression of BRCA1 and ZBRK1 genes, but inhibited the expression of p21 gene. Conclusion: It is confirmed that CtIP gene has the significantly inhibitory effect on injured cerebral endothelia cells, and the regulatory relationship between CtIP gene and BRCA1, ZBRK1 and p21 genes in the process of injury are determined.

The study on pathological mechanism and solution method for spinal cord ischemia reperfusion injury.

OBJECTIVE: We aimed at investigating the pathological mechanism changing of injury during reperfusion injury, reperfusion time correlation and compliance, finding the blood supply and improving the secondary damage. MATERIALS AND METHODS: A total of 180 patients who underwent a surgical procedure and that received normal saline intraperitoneally immediately after the patients' aortic occlusions were investigated. Patients were divided in three groups. Experimental conditions and programs were designed for various approaches. RESULTS: Thirty min after the onset of ischemia, we found a decrease in the local blood flow in the lumbar spinal cord, almost -77.48% of the baseline, which was reversed partially by initial reperfusion, even exceeding the baseline level. However, 1 hour after reperfusion, the blood flow was again decreased to the level below the baseline, followed by a decline to 207.13% ± 38.25 PU for 3 h without any recovery. Attenuating this secondary damage with neuroprotective strategies requires an understanding of these pathophysiologic processes. CONCLUSIONS: This study showed the pathological mechanism changes during reperfusion injury and reperfusion time correlation and compliance, and analyzed some of the important pathophysiologic processes involved in secondary damage after spinal cord injury. Moreover, our research discusses a number of pharmacologic therapies that have either been studied or have future potential for this devastating injury.

Epidemiological and clinical characteristics of severe fever with thrombocytopenia syndrome (SFTS) in China: an integrated data analysis.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease that was caused by a novel bunyavirus, SFTSV. The study aimed to disclose the epidemiological and clinical characteristics of SFTSV infection in China so far. An integrated clinical database comprising 1920 SFTS patients was constructed by combining first-hand clinical information collected from SFTS sentinel hospitals (n = 1159) and extracted data (n = 761) from published literature. The considered variables comprised clinical manifestations, routine laboratory tests of acute infection, hospitalization duration and disease outcome. SFTSV-IgG data from 19 119 healthy subjects were extracted from the published papers. The key clinical variables, case-fatality rate (CFR) and seroprevalence were estimated by meta-analysis. The most commonly seen clinical manifestations of SFTSV infection were fever, anorexia, myalgia, chill and lymphadenopathy. The major laboratory findings were elevated lactate dehydrogenase, aminotransferase, followed by thrombocytopenia, lymphocytopenia, elevated alanine transaminase and creatine kinase. A CFR of 12·2% was estimated, significantly higher than that obtained from national reporting data, but showing no geographical difference. In our paper, the mortality rate was about 1·9 parts per million. Older age and longer delay to hospitalization were significantly associated with fatal outcome. A pooled seroprevalence of 3·0% was obtained, which increased with age, while comparable for gender. This study represents a clinical characterization on the largest group of SFTS patients up to now. A higher than expected CFR was obtained. A wider spectrum of clinical index was suggested to be used to identify SFTSV infection, while the useful predictor for fatal outcome was found to be restricted.

¹H-MRS before and after resuscitation following selective cerebral ultra-profound hypothermic blood flow occlusion in monkeys.

We investigated the effect of selective cerebral ultra-profound hypothermic blood flow occlusion on brain tissue and cell metabolism to ascertain the efficacy and safety of selective deep hypothermic technologies using proton magnetic resonance spectroscopy ((1)H-MRS). The bilateral carotid artery was blocked at room temperature for 10 min. Other neck vessels were then blocked through cold perfusion of the internal carotid artery and reflux of the ipsilateral jugular vein. Thus, selective cerebral extracorporeal circulation was established. Brain temperature was reduced to 15.1° ± 0.9°C. After 60 min, cerebral blood flow recovered naturally. Routine magnetic resonance imaging (MRI), diffusion-weighted imaging (DWI), and (1)H-MRS examination of the bilateral frontal cortex and basal ganglia were performed prior to surgery and 4, 24, 72 h, 21 days after recovery. The formants and areas under the curve (AUC) of N-acetyl aspartate (NAA), choline (Cho), creatine/phosphocreatine (Cr/Cr2) were analyzed using 1H-MRS. The pre- and postoperative AUC of NAA and Cho at different time points were compared. Conventional MRI and DWI showed no abnormal signal changes in the brain parenchyma or right basal ganglia before and after surgery (P > 0.05). There was no significant difference in the ratio between NAA/(Cr+Cr2) and Cho/(Cr+Cr2) before and after surgery in the bilateral basal ganglia and frontoparietal regions of the cortex (P > 0.05). Quantitative (1)H-MRS showed that selective deep cerebral hypothermia significantly improved the brain's tolerance to ischemia and hypoxia. Our results could provide a better understanding of the efficacy and safety of selective deep hypothermia and blood flow occlusion.

Changes in hippocampal ultrastructure and vimentin expression in rhesus monkeys following selective deep hypothermia and blood occlusion.

Previous studies have shown that selective cerebral profound hypothermia combined with antegrade cerebral perfusion can improve resistance to cerebral hypoxia-ischemia in monkeys. The aim of this study was to observe the effect of selective cerebral profound hypothermia on the ultrastructure and vimentin expression in monkey hippocampi after severe cerebral ischemia. Eight healthy adult rhesus monkeys were randomly divided into two groups: profound hypothermia (N = 5) and normothermia (N = 3). Monkeys in the profound hypothermia group underwent bilateral carotid artery and jugular vein occlusion for 10 minutes at room temperature. Ringer's solution at 4°C was then perfused through the right internal carotid artery and out of the right jugular vein, maintaining the brain temperature below 18°C. Sixty minutes later, cerebral blood flow was restored. The normothermia group underwent all procedures with the exception that the Ringer's solution was 37°C during perfusion. All animals in the profound hypothermia group were successfully resuscitated. No significant abnormalities of hippocampal morphology or ultrastructure were observed. In contrast, no monkeys were alive after perfusion in the normothermia group and they had abnormal hippocampal morphology and ultrastructure to different extents. Vimentin expression in the hippocampus was significantly lower in the profound hypothermia group (47.88% ± 1.66) than the normothermia group (79.51% ± 1.00; P < 0.01). We conclude that selective cerebral profound hypothermia following 10-min occlusion of the bilateral common carotid arteries was able to downregulate vimentin expression in the hippocampus and protect it from severe cerebral ischemia.

Prokaryotic expression and immunoassay of grass carp reovirus capsid VP6 protein.

Grass carp reovirus (GCRV) of the genus Aquareovirus and the family Reoviridae causes a severe hemorrhagic disease in grass carp fingerlings in China. GCRV genome is composed of 11 double-stranded RNA segments, of which segment 8 encodes the major core capsid protein VP6. In this study, the VP6 gene following an RT-PCR-amplification from the GCRV 104 strain was cloned into an expression vector pET-32a to obtain pET-32(a)-VP6. The VP6 protein was expressed in Escherichia coli BL21 as a fusion protein of 64 kDa. After purification with the HisLink Spin Protein Purification System the VP6 protein was used to raise a specific polyclonal antibody in Balb/c mice. Presence of VP6 protein was proved in bacterial lysates containing VP6 fusion protein by Western blot analysis and in GCRV-infected CIK cells by immunofluorescent staining using polyclonal antibody. These results may be helpful in further studies of interactions between GCRV and cells and in preparation of an engineered vaccine against GCRV.

T1 relaxation times for viability evaluation of the engrafted and the native liver in a rat model of heterotopic auxiliary liver transplantation: a pilot study.

Following a heterotopic auxiliary liver transplantation, commonly used measurements are either invasive or non-indicative of individual viability of the coexisting engrafted and native livers. Magnetic resonance imaging (MRI) was therefore tested for its potential to monitor the post-transplant hepatic viability in a rat model. Thirteen Wistar rats were systematically evaluated with MRI and serum biochemical liver parameters. Post-transplant complications and the causes of animal death were identified by autopsy and histo-pathological examinations. The data of the healthy survivors were compared with those of the rats that developed complications. On MRI, the hepatic complications could be depicted in the individual livers. A specific pattern of signal evolution was found in the livers of the healthy survivors: the mean T1 relaxation times of the engrafted livers increased immediately after transplantation (476 +/- 64 ms, mean +/- standard deviation, pre-operative; 730 +/- 48 ms, week 1) and then declined steadily to a 3 month value of 489 +/- 246 ms, while, following a transient first rise (476 +/- 64 ms, pre-operative; 589 +/- 28 ms, week 1), the mean T1 value of the native livers increased again 4 weeks after surgery and reached a 3 month value of 859 +/- 43 ms. However, in the rats with various complications, the mean T1 relaxation times of the engrafted livers continued to increase throughout the first post-operative month (760 +/- 48 ms, week 1; 922 +/- 76 ms, week 4), while that of the native liver only varied mildly (546 +/- 25 ms, week 1; 473 +/- 25 ms, week 4). After the first post-transplant week, the healthy engrafted livers could already be distinguished from those with complications by a significant decrease in T1 relaxation times. These data suggest that, besides demonstrating major complications, MRI may allow one to monitor the viability of each liver by analysing the relative signal intensity and T1 relaxation times after a heterotopic auxiliary liver transplantation.

Effects of Re-arterialization on early graft function and regeneration in the rat model of heterotopic auxiliary liver transplantation.

In the rat model of heterotopic auxiliary liver transplantation, graft re-arterialization may influence the outcome of inter-liver competition. This was investigated in the current study using two transplanted groups with or without graft re-arterialization. Immediately after reperfusion, the re-arterialized grafts showed significantly higher bile flow rate and bilirubin excretion than the grafts without re-arterialization. DNA synthesis rate was also increased more drastically in the re-arterialized group following the transplantation. Without re-arterialization, the rats developed more pronounced cytolysis and cholestasis. Among the long-term survivors, all healthy re-arterialized grafts regenerated, whereas 5/6 non-re-arterialized grafts atrophied. These data demonstrate that the re-arterialization increases graft survival by improving early hepatic function, enhancing regenerative response and preventing post-transplant biliary complications in this rat model.

The need to handicap the recipient's native liver in the rat model of heterotopic auxiliary liver transplantation.

In the rat model of heterotopic auxiliary liver transplantation (HALTx), the opinion varies on whether and how the recipient's native liver should be handicapped. To avoid atrophy of the transplanted organ, in this study, two different handicaps were evaluated and their effects on post-operative animal survival and liver biology are described. With a sole portacaval shunt (group 1) all rats survived longer than 3 months. An additional handicap of the liver with either a 68% partial hepatectomy (68% PH) (group 2), or both a 68% PH and a common bile duct ligation (CBDL) (group 3) led to a 100% mortality within 2 days after surgery. When an auxiliary liver was transplanted to the rats handicapped with a 68% PH (group 4), serum Bilirubin and ALAT values were significantly lower than those handicapped with both a 68% PH and a CBDL (group 5). Autopsy and histology of the long-term survivors revealed the atrophy of the engrafted livers and the regeneration of the native livers in group 4, whereas it showed the opposite in group 5. Thus the various manipulations of the native liver do influence differently the post-transplant animal survival, serum liver biochemistry and the outcome of the engrafted liver in this rat model of HALTx.

Evaluation of graft viability in heterotopic auxiliary liver transplantation in the rat.

In the rat model of heterotopic auxiliary liver transplantation, the coexistence of the engrafted liver and the recipient's native liver makes it difficult to evaluate the posttransplant graft viability. In this study, auxiliary liver transplantation was performed in Wistar rats, in which the recipient's native liver was handicapped with a 68% partial hepatectomy and a common bile duct ligation. Serum biochemistry of the liver was analyzed and compared with that of the selected control group. The surgical handicap of the liver showed severe damaging effects: the handicapped native livers appeared atrophic at autopsy, and no long-term animal survival could be achieved without an auxiliary liver transplantation. As the engrafted liver corrected the cholestasis of the handicapped native liver, significant differences of serum biochemistry were found between the transplanted group and the control group: for bilirubin concentration and gamma glutamyl transferase activity from postoperative day 3 to 28 (p < .05); for alkaline phosphatase on days 3, 7, 14, and 28 (p < .05); for alanine aminotransferase activity on days 3 and 14 (p < .05); and for aspartate aminotransferase activity on day 14 (p < .05). The efficiency to induce hepatic failure and to hamper its regeneration capacity in the native liver makes animal survival and liver biology as reliable parameters to evaluate the posttransplant graft viability in this rat model.

Obstetric complications in young teenagers.

We compared pregnancy outcome in 286 teenaged primigravidas (less than or equal to 16 years old) and 267 adult primigravidas (21 to 25 years old) who had similar prenatal care, socioeconomic status, and racial balance. The incidence of preterm labor and delivery of a low birthweight infant was significantly higher in the teenagers. The incidence of preeclampsia was significantly higher in the adults. Cesarean delivery was not done more frequently in teenagers, nor was there a higher incidence of infants small for gestational age, anemia, and abnormal presentation in labor. The birthweight of infants of black teenagers was significantly lower than the birthweight of those of white teenagers, and overall birthweight was significantly lower in infants of teenagers than those of adults. Although prenatal care, socioeconomic factors, and racial balance were comparable for young teenagers and adults, teenagers were still at a significantly greater risk for delivery of smaller infants, preterm labor, and low birthweight infants.

Listeriosis as an obstetric complication in an immunocompromised patient.

We have reported a case of maternal death associated with Listeria monocytogenes septicemia in a woman who was being treated with immunosuppressive drugs for lupus nephritis. This report, coupled with a previous case of L monocytogenes sepsis in a pregnant patient with AIDS, emphasizes that L monocytogenes infection may be an important, unrecognized pathogen in pregnant women with impaired immunity.

Acute pyelonephritis in pregnancy.

There were 107 episodes of pyelonephritis associated with pregnancy or the early puerperium occurring in 103 gravidas investigated retrospectively for information concerning prematurity, low birthweight, and antibiotic susceptibility patterns in the recovered microorganisms. No difference was found in the incidence of prematurity on low birthweight between that group and a control group of gravidas from the same population. Members of the Enterobacteriaceae genus were the most common bacterial isolates from the urine, with a large portion of E. coli being resistant to both ampicillin (33%) and cephalothin (13%). Treated pyelonephritis associated with pregnancy does not appear to predispose to prematurity or low birthweight in this population. Also, initial therapy with a first-generation cephalosporin may no longer be appropriate, because a significant number of isolates (11%) were resistant to cephalothin.

Elevated mannose levels detected by gas-liquid chromatography in hydrolysates of serum from rats and humans with candidiasis.

We have developed a gas-liquid chromatographic method for identification of mannose in serum hydrolysates by utilizing peracetylated aldononitrile derivatives. Experimentally infected rats and human subjects with candidiasis were studied. Peaks in unknown samples were identified by co-chromatography of reference carbohydrates. Inositol was used as an internal standard. Mannose was identified in hydrolysates of normal rat (391.85 +/- 66.18 micrograms/ml) and human (297.87 +/- 77.81 micrograms/ml) sera. Significantly increased concentrations of mannose (greater than 526.21 micrograms/ml) were demonstrated in hydrolysates of sera from 26 of 36 (72%) experimentally infected rats tested 1 or more days after onset of infection. Significant increases (greater than 453.49 micrograms/ml) were also seen in humans with localized (6/15), transient (2/2), and disseminated (5/7) candidiasis. The concentrations were highest in patients with candidemia occurring either transiently or as part of disseminated infection. The concentrations in patients with localized candidiasis were lower but were still significantly greater than control. These data suggest that changes in mannose concentration, as measured, may serve as markers of candidiasis. The amount of mannose present may be in part a function of the extent of the infection.