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Background:. Endotoxin tolerance (ET) is a protective phenomenon in which pre-treatment with a tolerance dose of lipopolysaccharide (LPS) leads to dramatically elevated survival. Accumulating evidence has shown that peripheral T cells contribute to the induction of ET. However, what happens to T cell development in the thymus under ET conditions remains unclear. The purpose of this study was to analyze the alterations in thymocyte populations (double-positive [DP] and single-positive [SP] cells) under ET conditions. Methods:. Mice were intraperitoneally injected with LPS at a concentration of 5 mg/kg to establish an LPS tolerance model and were divided into two groups: a group examined 72 h after LPS injection (72-h group) and a group examined 8 days after LPS injection (8-day group). Injection of phosphate-buffered saline was used as a control (control group). Changes in thymus weight, cell counts, and morphology were detected in the three groups. Moreover, surface molecules such as CD4, CD8, CD44, CD69, and CD62L were analyzed using flow cytometry. Furthermore, proliferation, apoptosis, cytokine production, and extracellular signal-regulated kinase (ERK) pathway signaling were analyzed in thymocyte populations. The polymorphism and length of the T-cell receptor (TCR) β chain complementarity-determining region 3 (CDR3) were analyzed using capillary electrophoresis DNA laser scanning analysis (ABI 3730). Results:. Thymus weight and cell counts were decreased in the early stage but recovered by the late stage in a murine model of LPS-induced ET. Moreover, the proportions of DP cells (control: 72.130 ± 4.074, 72-h: 10.600 ± 3.517, 8-day: 84.770 ± 2.228), CD4+ SP cells (control: 15.770 ± 4.419, 72-h: 44.670 ± 3.089, 8-day: 6.367 ± 0.513), and CD8+ SP cells (control: 7.000 ± 1.916, 72-h: 34.030 ± 3.850, 8-day: 5.133 ± 0.647) were obviously different at different stages of ET. The polymorphism and length of TCR β chain CDR3 also changed obviously, indicating the occurrence of TCR rearrangement and thymocyte diversification. Further analysis showed that the expression of surface molecules, including CD44, CD69, and CD62L, on thymocyte populations (DP and SP cells) were changed to different degrees. Finally, the proliferation, apoptosis, cytokine production, and ERK pathway signaling of thymocyte populations were changed significantly. Conclusion:. These data reveal that alterations in thymocyte populations might contribute to the establishment of ET.

Background. Patients with temporal lobe epilepsy (TLE) originating from different seizure onset zones had distinct electrophysiological characteristics and surgical outcomes. In this study, we aimed to investigate the relationship between the origin and prognosis of TLE, and the stereoelectroencephalography (SEEG) features. Methods. Thirty patients with TLE, who underwent surgical treatment in our functional neurosurgery department from January 2016 to December 2017, were enrolled in this study. All patients underwent anterior temporal lobectomy after an invasive pre-operative evaluation with SEEG. Depending on the epileptic focus location, patients were divided into those with medial temporal lobe seizures (MTLS) and those with lateral temporal lobe seizures (LTLS). The Engel classification was used to evaluate operation effectiveness, and the Kaplan-Meier analysis was used to detect seizure-free duration. Results. The mean follow-up time was 25.7 ± 4.8 months. Effectiveness was 63.3% for Engel I (n = 19), 13.3% for Engel II, 3.3% for Engel III, and 20.0% for Engel IV. According to the SEEG, 60.0% (n = 18) had MTLS, and 40.0% (n = 12) had LTLS. Compared with the MTLS group, the operation age of those with LTLS was significantly greater (26.9 ± 6.9 vs. 29.9 ± 12.5 years, t = −0.840, P = 0.009) with longer epilepsy duration (11.9 ± 6.0 vs. 17.9 ± 12.1 years, t = −1.801, P = 0.038). Patients with MTLS had a longer time interval between ictal onset to seizure (67.3 ± 59.1 s vs. 29.3 ± 24.4 s, t = 2.017, P = 0.008). The most common SEEG ictal pattern was a sharp/spike-wave rhythm in the MTLS group (55.6%) and low-voltage fast activity in the LTLS group (58.3%). Compared with the LTLS group, patients with MTLS had a more favorable prognosis (41.7% vs. 77.8%, P = 0.049). Post-operative recurrence was more likely to occur within three months after the operation for both groups, and there appeared to be a stable long-term outcome. Conclusion. Patients with MTLS, who accounted for three-fifths of patients with TLE, showed a more favorable surgical outcome.

Background: Healthcare-associated pneumonia (HCAP) is associated with drug-resistant pathogens and high mortality, and there is no clear evidence that this is due to inappropriate antibiotic therapy. This study was to elucidate the clinical features, pathogens, therapy, and outcomes of HCAP, and to clarify the risk factors for drug-resistant pathogens and prognosis. Methods: Retrospective observational study among hospitalized patients with HCAP over 10 years. The primary outcome was 30-day all-cause hospital mortality after admission. Demographics (age, gender, clinical features, and comorbidities), dates of admission, discharge and/or death, hospitalization costs, microbiological results, chest imaging studies, and CURB-65 were analyzed. Antibiotics, admission to Intensive Care Unit (ICU), mechanical ventilation, and pneumonia prognosis were recorded. Patients were dichotomized based on CURB-65 (low- vs. high-risk). Results: Among 612 patients (mean age of 70.7 years), 88.4% had at least one comorbidity. Commonly detected pathogens were Acinetobacter baumannii, Pseudomonas aeruginosa, and coagulase-negative staphylococci. Initial monotherapy with β-lactam antibiotics was the most common initial therapy (50%). Mean age, length of stay, hospitalization expenses, ICU admission, mechanical ventilation use, malignancies, and detection rate for P. aeruginosa, and Staphylococcus aureus were higher in the high-risk group compared with the low-risk group. CURB-65 ≥3, malignancies, and mechanical ventilation were associated with an increased mortality. Logistic regression analysis showed that cerebrovascular diseases and being bedridden were independent risk factors for HCAP. Conclusion: Initial treatment of HCAP with broad-spectrum antibiotics could be an appropriate approach. CURB-65 ≥3, malignancies, and mechanical ventilation may result in an increased mortality.

BACKGROUND Although the use of hepatitis B immunoglobulin (HBIG) may lead to a significant reduction in recurrent hepatitis B virus (HBV) infection and improve the survival of patients who have undergone liver transplantation (LT) for hepatitis B-related diseases, the recurrence of the disease still remains at a lower level. Different clinical curative effects were observed in patients with the same HBV-related diseases and the same therapy. This study was undertaken to investigate whether the efficacy of HBIG is associated with FCGR3A gene polymorphisms in Chinese liver transplant patients. METHODS Altogether 77 patients who had received liver transplantation for hepatitis B-related diseases with more than one-year survival after surgery were studied. The recurrence of HBV was characterized by the appearance of HBsAg in serum after the operation. The FCGR3A genotyping was performed using genomic DNA sequencing (ABI 3037). Single nucleotide polymorphism at nucleotide 559 was detected by Polyphred. RESULTS Of the 77 patients, 14 were complicated with HBV recurrence post-transplant. The FCGR3A at nucleotide 559 TT was observed in 35 (45.5%) subjects, whereas TG in 31 (40.3%) and GG in 11 (14.3%). In the 559G carrier group (n = 42, 54.5%), the risk of HBV recurrence was 9.5%, and 1- and 2-year recurrence-free survival rates were 95.2% and 88.7%, respectively. In the 559G noncarrier group (n = 35, 45.5%), the risk of HBV recurrence was 28.6%, and 1- and 2-year recurrence-free survival rates were 74.3% and 69.3%, respectively. The risk of HBV recurrence and the recurrence-free survival rate were both statistically different between the 559G carrier and noncarrier groups (P < 0.05). CONCLUSIONS A single nucleotide polymorphism (T/G) at position 559 of the FCGR3A gene was found in Chinese patients. The efficacy of HBIG in prophylaxis of HBV recurrence after LT is associated with the gene polymorphism, so detecting FCGR3A genotypes can be a clinical reference of the HBIG administration.

BACKGROUND Angiopoietin-1 (Ang-1) is an endothelial-specific growth factor that can promote angiogenesis. Studies demonstrated that Ang-1 can inhibit apoptosis of umbilical endothelial cells, but so far little is known about its effects on apoptosis of microvascular endothelial cells. With the apoptotic model of murine-cerebral-derived microvascular endothelial cells (bEnd.3) induced by serum-free culture, we attempted to clarify the molecular mechanism of bEnd.3 apoptosis, particularly its relation to cytochrome C (Cyt C). METHODS The cultured microvascular endothelial cell strain, bEnd.3 cell, was employed. An apoptotic model of bEnd.3 was established by serum-free culture. Flow cytometry after Annexin labeling and PI staining were used to assess the apoptotic effects of Ang-1 on bEnd.3, and the expression of Bax/Bcl-2, caspase 8, caspase 3, and Cyt C were detected with Western blotting and ELISA. RESULTS The apoptotic rate of bEnd.3 cells after stimulation with Ang-1 (100 ng/L) in serum-free medium was significantly higher than that in control group. Ang-1 inhibited early-stage apoptosis more than late-stage apoptosis provided by propidium iodide (PI) and AnnexinV double staining. The inhibition of Ang-1 on bEnd.3 cell apoptosis was strengthened with the increase in concentration (0 - 400 ng/ml). Ang-1 could decrease the expression of Bax, caspase3 and 8, and increase that of Bcl-2. The results of ELISA indicated that Ang-1 significantly decreased CytC content in cytoplasm and increase that in mitochondria. CONCLUSIONS Ang-1 could inhibit bEnd.3 apoptosis induced by serum-free medium culture. The apoptosis was associated with decreased Bax expression, increased Bcl-2 expression, which result in Cyt C transferring from mitochondria to cytoplasm, and then caspases activation are reduced and cell apoptosis is suppressed.

by obstruction or rupture of the peritoneal or retroperitoneal lymphatic glands, is defined as the accumulation of chyle in the peritoneal cavity. It is a difficult management problem due to the serious mechanical, nutritional, and immunological consequences of the constant loss of protein and lymphocytes. Most investigators believe that the incidence of chylous ascites is increasing because of more aggressive thoracic and retroperitoneal surgery and also because of the prolonged survival of patients with cancer. Although the incidence of chylous ascites has increased in recent years, treatment remains unsatisfactory. The standard conservative treatments include paracentesis, a medium chain triglyceride (MCT) based diet, total parenteral nutrition (TPN), and recently recommended somatostatin, are preferable in most cases, but usually need several weeks to cure chyloperitonum. Open or laparoscopic ligation of ruptured lymph ducts, a commonly used procedure for persistent chylous ascites, may fail in patients without visible chylous leak. Fibrin glue, a widely used approach to treat various kinds of fistulae, has recently been reported to seal chylous leak during operation. Here we present two cases of chylous ascites who were successfully treated with fibrin glue in different settings.

There is no validated blood biomarker available for glioma management. Invasive growth is the key feature of glioma. We assessed the clinical usefulness of plasma tissue inhibitor of metalloproteinase 1 (TIMP-1), which has less molecular weight than metalloproteinases, as a potential blood biomarker for glioma.A total of 285 patients and 59 normal subjects were studied. Plasma concentration of TIMP-1 was measured with enzyme-linked immunosorbent assay. Plasma TIMP-1 was compared between normal and glioma patients, between patients with different pathological grades, and between patients with different prognoses. Longitudinal changes in plasma TIMP-1 during treatment were also evaluated. Plasma matrix metalloproteinase (MMP)-9 level was also assayed and its clinical usefulness was compared with that of TIMP-1.Plasma TIMP-1 and MMP-9 were both increased in glioma patients compared with normal controls (TIMP-1: P0.001; MMP-9: P = 0.007). Plasma TIMP-1 increases with increased tumor grade. In Grade IV gliomas, plasma TIMP-1 significantly increased after "successful removal" of the tumor (paired samples t-test, before operation vs. during chemotherapy without recurrence, t = -2.131, P = 0.038), but did not change significantly at the time of tumor recurrence (during chemotherapy without recurrence vs. after tumor recurrence, t = -0.652, P = 0.632). High plasma TIMP-1 level correlated with better survival in Grade IV glioma patients (hazard ratio: 0.550, 95% CI: 0.101-1.000, P = 0.036). In Grade IV gliomas, patients with higher plasma TIMP-1 had significantly longer survival time than those with lower plasma TIMP-1 level (25.23 vs. 18.95 months, log-rank P = 0.045). Plasma MMP-9 did not show significant association with either the pathological grade or the prognosis of glioma patients.Plasma TIMP-1 is associated with the diagnosis and prognosis of glioma patients. It appears to have better usefulness for guiding clinical decision making than plasma MMP-9. Further studies in an expanded patient population are needed to better define its clinical usefulness.

Chronic kidney disease (CKD) is a growing public health problem with well-established risk factors. Other contributing factors, however, remain to be identified. Systemic inflammation in asthma plays a significant role in the development of other diseases. We therefore initiated a study to assess whether the growing prevalence of asthma is associated with an increase in the risk of CKD.We conducted a retrospective cohort study using data from 3015 patients with asthma aged 14 years and older who were registered and followed up in Asthma Control Study at the Department of Respiratory Medicine of three medical centers from 2005 to 2011. History, asthma control test (ACT), and asthma stage were used to assess the traits of asthma. CKD was defined as proteinuria and/or reduced estimated glomerular filtration rate (eGFR) (60 ml×min(-1)×1.73 m(-2)) in two consecutive follow-up surveys. We used logistic regression models, adjusting for age, sex, and other confounding factor to determine associations between the traits of asthma and CKD. Kaplan-Meier curves were used to analyze patient outcomes.A total of 2354 subjects with complete data were recruited for this study with mean age (45.4±10.4) years. After 6 years of follow-up, 9.6% (n = 227) of the analytic cohort developed proteinuria and 3.1% (n = 72) progressed to eGFR60 ml×min(-1)×1.73 m(-2). The patients with20 years asthma history, not well-controlled or persistent asthma patients had higher incidence of proteinuria and reduced eGFR compared with patients with ≤20 years asthma history, at least well-controlled or remission asthma, respectively. The multivariable adjusted OR for proteinuria and reduced eGFR in participants with persistent asthma was 1.49; (95% confidence interval (CI) 1.17-1.91) and 2.07 (95% CI 1.34-4.42). Compared to patients with no asthma traits, there was a significant risk (OR, 3.39; 95% CI 1.36-8.73) for those who met all three traits, including asthma history20 years, not well-controlled and persistent stage, after adjusting for potential confounding factors.In this retrospective cohort study, we found that persistent asthma was associated with an increased risk of CKD, which was independent of obesity, diabetes, hypertension, and other well-established risk factors. Future studies should be directed to elucidate the mechanisms underlying the association between asthma and CKD.

Folate plays a critical role in nucleotide synthesis and DNA methylation, and was considered to be associated with anti-carcinogenesis.from studies that concern the relationship between the folate intake or serum folate levels and lung cancer risk showed no consistency, which requires our further comprehensive meta-analysis.Systematic literature search was conducted to identify the relevant studies (published prior to February 2013) according to standard protocol. Estimated effects were calculated under both random-effects and fixed-effects models. Heterogeneity between studies and publication bias were also evaluated.A total of 4390 cases and 6138 controls from 6 case-control studies revealed a significant overall inverse association between folate intake and lung cancer risk (OR = 0.74, 95%CI = 0.65 - 0.84, P0.001). Summary of 1438 cases and 2582 controls from 4 case-control studies and 44 cases out of a cohort of 1988 participants suggested a marginal association without significance (OR = 0.78, 95%CI = 0.60 - 1.02, P = 0.075) between high serum folate levels and less lung cancer susceptibility; however, subgroup analysis about population-based case-control studies showed that high serum folate levels significantly associated with the reduced lung cancer risk (OR = 0.76, 95%CI = 0.58 - 1.00, P = 0.048).Higher folate intake can be a protective factor against lung cancer risk, and higher serum folate level is probably associated with reduced lung cancer risk in marginal manner, though more studies are warranted to confirm these associations.

Recurrence of hepatitis B-related hepatocellular carcinoma (HCC) after curative resection is the leading factor influencing the prognosis of the disease. Therefore, further improvement of long-term survival may depend on the prevention and treatment of the recurrent tumor. The aim of this research was to investigate the role of antiviral therapy and postoperative transcatheter arterial chemoembolization (TACE) in the prevention and treatment of hepatitis B-related HCC recurrence.One hundred and twenty patients who underwent curative resection of hepatitis B-related HCC between January 2005 and June 2008 at our hospital were enrolled. Patients were divided into four groups according to the post-operative adjuvant therapy they received, i.e., control, antiviral therapy group, TACE group, and combined group. The disease-free survival (DFS) and the 12-, 24-, 36-month cumulative recurrence rates were studied.There was no significant difference between isolated postoperative antiviral therapy group and control in terms of disease-free survival (P = 0.283), while it was significantly higher in the TACE group compared to control (P = 0.019). In all patients, however, viral prophylactic therapy combined with/without TACE brought a favorable result compared to those only with/without TACE (P0.001). Similarly, no matter combined with or without antiviral treatment, postoperative TACE prolonged DFS (P = 0.015). Naturally, a combination of viral prophylactic therapy on the baseline TACE significantly benefited patients' postoperative DFS (P = 0.047) and vice verse (P = 0.002). The 24-month cumulative recurrence rates of combined group were significantly lower than that of isolated control group and antiviral therapy (P0.001 and P = 0.011 respectively). However, 36-month recurrence rate was significantly different in the control group compared to the TACE group and combined group (P = 0.040 and 0.002 respectively); same as the antiviral group compared to the combined group (P = 0.034).Post-operative TACE prevents early recurrence while antiviral therapy prevents late recurrence of HCC. Combination of antiviral therapy and TACE are suggested for prevention in HCC patients with high risk of recurrence.

Diabetes has been associated with increased risk of fracture and impaired fracture healing. The aim of this study was to examine the influence of diabetes on perioperative complications, length of stay and ambulatory ability recovery in individuals with hip fracture, and to determine whether changes could be made to improve treatment outcome.The study included 707 hip fracture patients treated at Beijing Jishuitan Hospital between July 2009 and December 2010. The medical history and perioperative complications were compared between non-diabetic and diabetic groups. Length of stay, days awaiting surgery, and days of hospitalization after surgery were also analyzed. Ambulatory ability was compared at 1-year follow-up using the Chi-square test and Fisher's exact test. An independent Student's t-test was used to compare normally distributed continuous data.Patients with diabetes were more likely than non-diabetic patients to develop cardiac perioperative complications (8.9% vs. 3.0%, P = 0.021), urinary tract infections (12.0% vs. 2.8%, P0.001), and gastrointestinal symptoms (15.0% vs. 6.8%, P = 0.003). No difference in perioperative complications was observed between the groups. Days awaiting surgery and length of hospital stay were both longer in the diabetic group ((8.0 ± 5.1) vs. (6.2 ± 3.7) days and (16.5 ± 3.8) vs. (13.3 ± 3.8) days, P0.001, respectively). Before the occurrence of fracture, patients with diabetes were less likely to be ambulatory outdoors (71.9% vs. 85.9%, P0.001) and had more restricted walking ability. After at least 1-year follow-up, similar proportions of patients in the non-diabetic and diabetic groups (16.1% and 15.9%, respectively), who were able to ambulate outdoors before the fracture, became housebound till the final follow-up.Diabetics are at increased risk of specific complications and have a longer time to surgery and longer in-hospital stay, but generally have similar recovery to non-diabetics thereafter.

Although traumatic brain injury can lead to opening the blood-brain barrier and leaking of blood substances (including water) into brain tissue, few studies of brain antigens leaking into the blood and the pathways have been reported. Brain antigens result in damage to brain tissues by stimulating the immune system to produce anti-brain antibodies, but no treatment has been reported to reduce the production of anti-brain antibodies and protect the brain tissue. The aim of the study is to confirm the relationship between immune injury and arachnoid granulations following traumatic brain injury, and provide some new methods to inhibit the immune injury.In part one, methylene blue was injected into the rabbits' cisterna magna after traumatic brain injury, and concentrations of methylene blue and tumor necrosis factor (TNF)-α in blood were detected to determine the permeability of arachnoid granulations. In part two, umbilical cord mesenchymal stem cells and immature dendritic cells were injected into veins, and concentrations of interleukin 1 (IL-1), IL-10, interferon (IFN)-γ, transforming growth factor (TGF)-β, anti-brain antibodies (ABAb), and IL-12 were measured by ELISA on days 1, 3, 7, 14 and 21 after injury, and the numbers of leukocytes in the blood were counted. Twenty-one days after injury, expression of glutamate in brain tissue was determined by immunohistochemical staining, and neuronal degeneration was detected by HE staining.In part one, blood concentrations of methylene blue and TNF-α in the traumatic brain injury group were higher than in the control group (P0.05). Concentrations of methylene blue and TNF-α in the trauma cerebrospinal fluid (CSF) injected group were higher than in the control cerebrospinal fluid injected group (P0.05). In part two, concentrations of IL-1, IFN-γ, ABAb, IL-12, expression of glutamate (Glu), neuronal degeneration and number of peripheral blood leukocytes were lower in the group with cell treatment compared to the control group. IL-10 and TGF-β were elevated compared to the control group.Traumatic brain injury can lead to stronger arachnoid granulations (AGs) permeability; umbilical cord mesenchymal stem cells and immature dendritic cells can induce immune tolerance and reduce inflammation and anti-brain antibodies to protect the brain tissue.

Neuroblastoma (NB) is one of the most common malignant solid tumors of childhood. It is still not clear whether the apoptosis of tumor cells or the non-tumor cells contributes to the increase of concentration of cytochrome c (Cyt c) in the serum of the cancer patients. The aim of this research was to identify the source of the Cyt c in the serum when the tumor grows up by subcutaneous inoculation of human NB cells into nude mice.We subcutaneously inoculated human NB cells (KP-N-NS) into nude mice and collected the sera of tumor-bearing mice (n = 14) and control mice (n = 25) 4 weeks later in order to screen for and identify differentially expressed proteins in the serum. Differentially expressed proteins in the serum were screened by surface-enhanced laser desorption/ionization-time-of-flight (SELDI-TOF) mass spectrometry.The relative intensity of a protein having a mass-to-charge ratio (m/z) of 11 609 was 3338.37 ± 3410.85 in the tumor group and 59.84 ± 40.74 in the control group, indicating that the expression level of this protein in the tumor group was 55.8 times higher than that in the control group. Serum proteins were separated and purified by high-performance liquid chromatography (HPLC). Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) was performed to produce peptide mass fingerprints (PMFs). Spectrum analysis and a database search revealed that the highly expressed protein (m/z = 11 605.4) from the serum of tumor-bearing mice was the mouse Cyt c.Increased concentration of Cyt c in the serum of tumor-bearing nude mice might be partially attributed to the secretion of this protein by non-tumor cells.

Previous studies have shown that glioma patients have lower blood IgE levels than controls. To evaluate its potential as a surrogate biomarker for glioma, we measured plasma IgE levels in glioma patients and healthy controls, and correlated them with clinicopathological factors and the patients' outcome.We used enzyme-linked immunosorbant assay (ELISA) to determine the plasma IgE levels of 25 normal subjects and 252 glioma patients (85 patients with grade II glioma, 46 patients with grade III glioma, and 121 patients with glioblastoma). We also collected longitudinal plasma samples from glioblastoma patients and compared the plasma IgE levels before operation, one week after operation, in the middle of radiotherapy, after two cycles of chemotherapy, and after recurrence. The correlations between plasma IgE levels and the outcomes of the patients were determined.Plasma IgE levels were significantly lower in glioma patients (P = 0.004); patients with low-grade glioma have lower IgE levels than patients with high-grade glioma do (P = 0.029). In 24 patients with both preoperative plasma and two-cycle chemotherapy plasma samples, IgE levels increased after successful removal of the tumor (P = 0.021), and the increase correlated with the patients' survival (increase100 ng/ml vs. ≤ 100 ng/ml, 127.5 weeks vs. 62.3 weeks. P = 0.012, log-rank). Plasma IgE level increase of100 ng/ml has a specificity of 80% and a sensitivity of 78% to predict the patients' long survival (18 months).Our results suggest that plasma IgE level correlates with clinical and pathological factors in glioma patients. It has the potential to be a biomarker for glioma patients.

p73, a homologue of p53, has been located at chromosome 1p36-33, a region of frequently observed loss of heterozygosity in breast cancers. The objective of the present study was to investigate the function of p73 in Japanese with breast cancers.Sixty Japanese patients with breast cancer were assessed by polymerase chain reaction single strand confirmation polymorphism analysis and direct sequencing to detect the p73 allele. p73 mRNA levels were also determined in 40 out of 60 patients by reverse-transcriptional polymerase chain reaction.We analyzed the entire open reading frame of the p73 gene by polymerase chain reaction single strand confirmation polymorphism and sequencing, and failed to identify any mutations of p73 in the encoding regions detected. Loss of heterozygosity of p73 was infrequent and only found in 9% of breast carcinomas. We revealed a few polymorphisms with a frequency of 13% - 29%, which had been reported previously. Down-regulation of p73 mRNA expression was observed in tumor tissues in comparison to the normal breast tissues. A significant inverse correlation was found between p73 transcripts and high histological grade, suggesting that down-regulated p73 expression could be related to poor prognosis in those patients.Our results suggest that p73 may serve as a tumor suppressor gene and its expression plays a role in tumorigenesis in Japanese patients with breast cancer.

Infection due to pandrug-resistant Pseudomonas aeruginosa (PDRPA) has become a challenge in clinical practice. The aim of this research was to summarize the treatment of large-area burns (60% - 80%) with PDRPA infection and respiratory failure in our hospital over the last two years, and to explore a feasible treatment protocol for such patients.We retrospectively analyzed the treatment of five patients with large-area burns accompanied by PDRPA infection and respiratory failure transferred to our hospital from burn units in hospitals in other Chinese cities from January 2008 to February 2010. Before PDRPA infection occurred, all five patients had open wounds with large areas of granulation because of the failure of surgery and dissolving of scar tissue; they had also undergone long-term administration of carbapenems. This therapy included ventilatory support, rigorous repair of wounds, and combined antibiotic therapy targeted at drug-resistance mechanisms, including carbapenems, ciprofloxacin, macrolide antibiotics and β-lactamase inhibitors.Four patients recovered from burns and one died after therapy.First, compromised immunity caused by delayed healing of burn wounds in patients with large-area burns and long-term administration of carbapenems may be the important factors in the initiation and progression of PDRPA infection. Second, if targeted at drug-resistance mechanisms, combined antibiotic therapy using carbapenems, ciprofloxacin, macrolide antibiotics and β-lactamase inhibitors could effectively control PDRPA infection. Third, although patients with large-area burns suffered respiratory failure and had high risks from anesthesia and surgery, only aggressive skin grafting with ventilatory support could control the infection and save lives. Patients may not be able to tolerate a long surgical procedure, so the duration of surgery should be minimized, and the frequency of surgery increased.

As one of the most commonly diagnosed diseases, chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is characterized by a variety of complex symptoms. Anxiety and depression are two of the most prevalent clinical manifestations of patients with CP/CPPS, and have adverse effects on the health of the subjects and prognosis of comorbidities. Such psychological disorders, however, have not been deeply and thoroughly studied in China. The aim of this study was to investigate the prevalence and severity of psychological disorders in Chinese adults with CP/CPPS.From April 2008 to June 2009, 80 patients and 40 age-matched healthy men participating in a voluntary health examination were recruited. The majority of the subjects completed the questionnaires on the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) as well as the hospital anxiety and depression scale (HADS).Of all the participants, 77 (96.3%) patients and 37 (92.5%) healthy controls completed the questionnaires. The average NIH-CPSI total score was 21.0 ± 9.5 for the patients and 2.2 ± 1.5 for the controls (P = 0.03). Of the 77 patients with CP/CPPS, 48 (62.3%), 5 (6.5%), and 1 (1.2%) had anxiety symptoms, depression symptoms, or both anxiety and depression symptoms, respectively. For the controls, the average HADS anxiety and depression scores in patients were 14.5 ± 6.8 and 5.2 ± 4.5, which were both significantly higher than in controls. Moreover, the prevalence and the symptom scores of both the HADS anxiety and depression were higher for the younger age group (35 years) than for the older age group (35 years).This preliminary study revealed that male patients with CP/CPPS had a higher prevalence of psychological disorders than in the control subjects. Moreover, the differences of the prevalence and severity of the psychological symptoms between the two different age groups may imply that psychological disorders related to CP/CPPS may be relieved with increasing age. The present study indicated that psychological evaluation is important in men with CP/CPPS, especially in younger men.

The cytotoxicity of dermal substitutes may be increased by the very processes used to deplete the cells. The present research aimed to investigate the method for monitoring the cytotoxicity of cell-free dermal substitutes using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) method.The cytotoxicity of four dermal substitutes was evaluated using the MTT method according to the standards set by the Chinese State Food and Drug Administration (SFDA). Swine acellular dermal matrix (SADM) and goat acellular dermal matrix (GADM) were produced using a repeated freeze-thaw method. Human dermal matrix glutaraldehyde composite (HADM-G) and SADM cross-linked with glutaraldehyde (SADM-G) were produced using conventional methods.The cytotoxicity of all dermal substitutes ranged from Grade 0 to Grade 1, meeting the standards of the Chinese FDA. The OD(490) of both SADM and GADM was higher than that of either HADM-G or SADM-G (P0.05).Dermal substitutes produced by the freeze-thaw method are less cytotoxic than those produced using conventional methods.

Pachymic acid (PA), a natural triterpenoid, is known to significantly reduce cell proliferation and induce apoptosis in vitro through initiation of mitochondria dysfunction. However, its effect on immune cells and anti-rejection following organ transplantation remains unknown.In this study, we investigated PA as a treatment to control acute rejection occurred in rats which had accepted cardiac transplantation. We measured apoptosis of peripheral blood lymphocyte (PBLs), and CD4(+) lymphocyte, as well as the number of CD4(+) and CD8(+) lymphocytes and the effect of PA on acute rejection in rats 7 days after cardiac transplantation.PA treatment might decrease allograft rejection, protect PBLs from apoptosis, and reduce the percentage of CD8(+) lymphocyte. PA neither regulated the number nor the apoptosis rate of CD4(+) lymphocyte.Our findings indicated that PA has an anti-apoptotic effect acting on PBLs through a novel mechanism involving stabilization of the PBLs mitochondrial transmembrane potential, an anti-rejection effect in rats after cardiac transplantation and an inhibiting effect to CD8(+) lymphocyte.

Although etomidate is associated with very few cardiovascular side-effects and minimal histamine release, it has a less inhibitory effect on the pharyngolaryngeal reflex. Hence, blunting the responses to endotracheal intubation is more dependent of opioids for etomidate-based anesthetic induction. This prospective, randomized, double-blinded study was designed to investigate the effects of low dose remifentanil, fentanyl or sufentanil on etomidate induction with respect to hemodynamics, conscious level changes and drug consumption.Ninety unpremedicated and normotensive patients with American Society of Anesthesiologists (ASA) physical status I or II undergoing elective major abdominal surgery were randomly assigned in a double blinded fashion to each of the three groups: groups F, R and S. A bolus dose of fentanyl 1 microg/kg, sufentanil 0.1 microg/kg or remifentanil 1 microg/kg was given over 60 seconds in groups F, S and R, respectively. In each instance this loading dose was followed by a continuous infusion (0.1, 0.01 or 0.1 microg x kg(-1) x min(-1) of fentanyl, sufentanil or remifentanil, respectively). After 5 minutes from start of opioid infusion, etomidate was titrated at a rate of 20 mg/min to a decrease in bispectral index (BIS) to 50. The time from administration of etomidate to loss of eyelash reflex or to a decrease in BIS to 50 was recorded. The blood pressure and heart rate were also recorded at different five time points. The average maximum percent changes of systolic blood pressure (|maximal or minimal measuring value-baseline|/baseline x 100%) were calculated.The time and the dosage of etomidate necessary to loss consciousness were greater in group F ((70.0 +/- 15.6) seconds; (0.35 +/- 0.05) mg/kg) than in groups S ((52.3 +/- 15.9) seconds; (0.26 +/- 0.06) mg/kg) and R ((56.2 +/- 20.2) seconds; (0.27 +/- 0.07) mg/kg) (P0.01). The three groups took similar time and amount of etomidate to achieve an adequate depth anesthesia (BIS = 50). The average maximum changes of systolic blood pressure were significantly different among the three groups: F, (25 +/- 6)% vs R, (13 +/- 4)% or S, (12 +/- 5)% (P0.001). The endotracheal intubation caused marked increases in blood pressure and heart rate in groups F and S, but not in group R, respectively (P0.01). The great hemodynamic changes occurred more frequently in group F than in groups R and S (P0.01). The incidence of heart rate decreases of more than 30% of the baselines after induction was higher in group R compared with groups F and S (P0.01).In normotensive and unpremedicated young adult patients receiving etomidate induction, low dose remifentanil or sufentanil significantly reduced the time and the amount of etomidate taken to loss unconsciousness compared with low dose fentanyl, but similar time interval and doses of etomidate were required to acquire adequate depth of anesthesia (BIS = 50) for these three opioids. Remifentanil was more effective in blunting the cardiovascular responses to endotracheal intubation, nevertheless, accompanying significant lower heart rate after induction.

Dysembryoplastic neuroepithelial tumor (DNT) is a rare benign neoplasm of the central nervous system affecting young people. A correct preoperative diagnosis is helpful for planning surgical strategies and improving prognosis. The purpose of this study was to characterize DNTs using magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) and to analyze the value of these two techniques in the diagnosis of DNTs.MR images of 13 patients with DNTs were reviewed retrospectively; and five of the patients also underwent MRS. Tumors were confirmed by surgery. The distribution, extension and signal features of the lesions were assessed, and the MRS results were analyzed.All tumors were supratentorial. The cortex was the main area involved, with nine tumors located in the temporal lobe, three in the frontal lobe, and one on the boundary between the temporal and occipital lobes. All cases had decreased signal intensity on T1-weighted MR images and increased signal intensity on T2-weighted images. On fluid attenuated inversion recovery weighted images, the hyperintense "ring sign" and internal septation of the lesion were seen in 9 cases. Eight tumors had well-demarcated borders. Peritumoral edema or mass effect was absent in all cases. A contrast enhancement examination was performed in 9 cases. Contrast enhancement was absent in five cases, and four cases showed significant enhancement. The MRS showed a low N-acetylaspartate peak and a lack of elevated choline-containing component (Cho) or Cho-Cr ratio (Cho/Cr) in five patients.The MRI findings of DNTs were stereotypical. The combination of MRI and MRS techniques were helpful in making a correct presurgical diagnosis.

As an alternative to open aneurysm repair, endovascular aortic repair (EVAR) has been applied to ruptured abdominal aortic aneurysm (rAAA). The aim of this study was to evaluate the immediate and long-term outcomes of EVAR for rAAA.From July 1997 to September 2007, 20 men and six women with rAAA (median age, 68 years) were treated with EVAR. Most patients with suspected rAAA underwent emergency computed tomographic angiography (CTA). The procedure was performed under general or local anesthesia. Endovascular clamping was attempted in hemodynamically unstable patients. Bifurcated endografts and aorto-uni-iliac (AUI) endografts with crossover bypass were used. Patients had CT scan prior to discharge, 3, 6, 12 months after discharge, and annually thereafter.Time between diagnosis and EVAR ranged from 1 hour to 5 days. EVAR was performed under general anesthesia in 21 patients, and under local anesthesia in five patients. Endovascular aortic clamping was performed in four patients. There was no conversion to open surgery during EVAR. Stent-graft insertion was successful in all patients. One patient died during EVAR from acute myocardial infarction. Ten patients had systolic blood pressure80 mm Hg. Eleven patients received a blood transfusion. Mean aneurysm size was (47 +/- 12) mm. Mean ICU stay was (8 +/- 3) days, mean hospital stay (18+/- 6) days, and mean procedure time (120 +/- 32) minutes. The 30-day mortality was 23% (6/26 patients), and major morbidity 35% (9/26 patients). Early endoleak occurred in 8/26 patients (31%). The mean follow-up was (18 +/- 7) months. No patient demonstrated migration of the stent-graft.EVAR is a safe and effective option for treatment of acute rAAA, independent of the patient's general condition. Immediate and mid-term outcomes are favorable, but long-term outcome is unknown. Multi-center studies are necessary to establish the role of EVAR for rAAA.

The level of basic fibroblast growth factor (bFGF) increases rapidly after cerebral ischemia. However, the molecular mechanisms for the effects of bFGF on cerebral microvascular endothelial cells (cMVECs) have not yet been fully elucidated. In this study, a murine cMVEC line, bEnd.3, was employed to study the effects of bFGF on cyclooxygenase (COX) expression and its downstream effects in cMVECs.After treatment with bFGF, RT-PCR and Western blotting analyses were carried out to evaluate the changes in COX-2 mRNA and protein expression, respectively. MTT assays were performed to measure cell proliferation. The prostaglandin E2 (PGE2) and vascular endothelial growth factor (VEGF) concentrations in the culture medium were measured by enzyme-linked immunosorbent assay (ELISA).COX-2 mRNA and protein expressions in bEnd.3 cells were induced by bFGF in time- and dose-dependent manners. The bFGF-induced COX-2 upregulation led to enhanced PGE2 production by bEnd.3 cells, and this effect was abolished by the selective COX-2 inhibitor NS-398. bFGF also increased VEGF production by bEnd.3 cells, and this effect was blocked by NS-398 and the EP1/2 (PGE2 receptors) antagonist AH6809. Furthermore, exogenous PGE2 increased VEGF production in bEnd.3 cells, and AH6809 blocked this effect.bFGF increases VEGF production in an autocrine manner by increasing COX-2-generated PGE2 in cMVECs and subsequently stimulates MVEC proliferation and angiogenesis.

Background The large phage antibody library is used to obtain high-affinity human antibody, and the Loxp/cre site-specific recombination system is a potential method for constructing a large phage antibody library. In the present study, a phage antibody library vector pDF was reconstructed to construct diabody more quickly and conveniently without injury to homologous recombination and the expression function of the vector and thus to integrate construction of the large phage antibody library with the preparation of diabodies. Methods scFv was obtained by overlap polymerase chain reaction (PCR) amplification with the newly designed VL and VH extension primers. loxp511 was flanked by VL and VH and the endonuclease ACC Ⅲ encoding sequences were introduced on both sides of loxp511. scFv was cloned into the vector pDF to obtain the vector pDscFv. The vector expression function was identified and the feasibility of diabody preparation was evaluated. A large phage antibody library was constructed in pDscFv. Several antigens were used to screen the antibody library and the quality of the antibody library was evaluated. Results The phage antibody library expression vector pDscFv was successfully constructed and confirmed to express functional scFv. The large phage antibody library constructed using this vector was of high diversity. Screening of the library on 6 antigens confirmed the generation of specific antibodies to these antigens. Two antibodies were subjected to enzymatic digestion and were prepared into diabody with functional expression. Conclusions The reconstructed vector pDscFv retains its recombination capability and expression function and can be used to construct large phage antibody libraries. It can be used as a convenient and quick method for preparing diabodies after simple enzymatic digestion, which facilitates clinical trials and application of antibody therapy. Chin Med J 2007;120(22):2011-2016

Although the use of hepatitis B immunoglobulin (HBIG) may lead to a significant reduction in recurrent hepatitis B virus (HBV) infection and improve the survival of patients who have undergone liver transplantation (LT) for hepatitis B-related diseases, the recurrence of the disease still remains at a lower level. Different clinical curative effects were observed in patients with the same HBV-related diseases and the same therapy. This study was undertaken to investigate whether the efficacy of HBIG is associated with FCGR3A gene polymorphisms in Chinese liver transplant patients.Altogether 77 patients who had received liver transplantation for hepatitis B-related diseases with more than one-year survival after surgery were studied. The recurrence of HBV was characterized by the appearance of HBsAg in serum after the operation. The FCGR3A genotyping was performed using genomic DNA sequencing (ABI 3037). Single nucleotide polymorphism at nucleotide 559 was detected by Polyphred.Of the 77 patients, 14 were complicated with HBV recurrence post-transplant. The FCGR3A at nucleotide 559 TT was observed in 35 (45.5%) subjects, whereas TG in 31 (40.3%) and GG in 11 (14.3%). In the 559G carrier group (n = 42, 54.5%), the risk of HBV recurrence was 9.5%, and 1- and 2-year recurrence-free survival rates were 95.2% and 88.7%, respectively. In the 559G noncarrier group (n = 35, 45.5%), the risk of HBV recurrence was 28.6%, and 1- and 2-year recurrence-free survival rates were 74.3% and 69.3%, respectively. The risk of HBV recurrence and the recurrence-free survival rate were both statistically different between the 559G carrier and noncarrier groups (P0.05).A single nucleotide polymorphism (T/G) at position 559 of the FCGR3A gene was found in Chinese patients. The efficacy of HBIG in prophylaxis of HBV recurrence after LT is associated with the gene polymorphism, so detecting FCGR3A genotypes can be a clinical reference of the HBIG administration.

Intubating laryngeal mask airway (ILMA) offers a new approach for orotracheal intubation and is expected to produce less cardiovascular stress responses. However, the available studies provide inconsistent results. The purpose of this study was to identify whether there is a clinically relevant difference in hemodynamic responses to orotracheal intubation by using ILMA and direct laryngoscope (DLS).A total of 53 adult patients, ASA physical status I-II, scheduled for elective plastic surgery under general anesthesia requiring the orotracheal intubation, were randomly allocated to either DLS or ILMA groups. After a standard intravenous anesthesia induction, orotracheal intubation was performed. Noninvasive blood pressure and heart rate were recorded before (baseline values) and after anesthesia induction (post-induction values), at intubation and every minute for the first 5 minutes after intubation. The data were analyzed using Chi-square test, paired and unpaired Student's t test, and repeated-measures analysis of variance as appropriate.The mean intubation time in the ILMA group was longer than that in the DLS group (P0.05). The blood pressure and heart rate increased significantly after intubation in the two groups compared to the post-induction values (P0.05), but the maximum value of blood pressure during the observation did not exceed the baseline value, while the maximum value of heart rate was higher than the baseline (P0.05). During the observation, there were no significant differences in blood pressure and heart rate among each time point and in the maximum values between the two groups.Orotracheal intubations by using ILMA and DLS produce similar hemodynamic response. ILMA has no advantage in attenuating the hemodynamic responses to orotracheal intubation compared with DLS.

Angiopoietin-1 (Ang-1) is an endothelial-specific growth factor that can promote angiogenesis. Studies demonstrated that Ang-1 can inhibit apoptosis of umbilical endothelial cells, but so far little is known about its effects on apoptosis of microvascular endothelial cells. With the apoptotic model of murine-cerebral-derived microvascular endothelial cells (bEnd.3) induced by serum-free culture, we attempted to clarify the molecular mechanism of bEnd.3 apoptosis, particularly its relation to cytochrome C (Cyt C).The cultured microvascular endothelial cell strain, bEnd.3 cell, was employed. An apoptotic model of bEnd.3 was established by serum-free culture. Flow cytometry after Annexin labeling and PI staining were used to assess the apoptotic effects of Ang-1 on bEnd.3, and the expression of Bax/Bcl-2, caspase 8, caspase 3, and Cyt C were detected with Western blotting and ELISA.The apoptotic rate of bEnd.3 cells after stimulation with Ang-1 (100 ng/L) in serum-free medium was significantly higher than that in control group. Ang-1 inhibited early-stage apoptosis more than late-stage apoptosis provided by propidium iodide (PI) and AnnexinV double staining. The inhibition of Ang-1 on bEnd.3 cell apoptosis was strengthened with the increase in concentration (0 - 400 ng/ml). Ang-1 could decrease the expression of Bax, caspase3 and 8, and increase that of Bcl-2. The results of ELISA indicated that Ang-1 significantly decreased CytC content in cytoplasm and increase that in mitochondria.Ang-1 could inhibit bEnd.3 apoptosis induced by serum-free medium culture. The apoptosis was associated with decreased Bax expression, increased Bcl-2 expression, which result in Cyt C transferring from mitochondria to cytoplasm, and then caspases activation are reduced and cell apoptosis is suppressed.

A living fetus within the maternal uterus provides an example of allogene tolerance in mammals. Poria cocos Wolf is the main component of many Chinese medicinal combination drugs that have therapeutic effects on recurrent spontaneous abortion and that can maintain pregnancy until delivery. It was hypothesized that this herbal medicine can also prolong allograft survival after organ transplantation. Here, in an in vivo study, we report the anti-rejection effect of the ethanol extract of Poria cocos Wolf (EEPCW) in rats after cardiac allograft implantation.Ten normal rats were healthy controls. Eighty rats receiving homologous heart transplants were divided into 4 groups of 20 rats each based on type of treatment: olive oil 8 ml.kg(-1).d(-1), EEPCW 25 mg.kg(-1).d(-1), EEPCW 50 mg.kg(-1).d(-1) or cyclosporin A 5 mg.kg(-1).d(-1). Allograft survival was observed in 10 rats from each group. On the seventh day post transplantation, pathological lesions and percentages of CD3+, CD4+, and CD8+ lymphocytes and the CD4+/CD8+ ratio in peripheral blood were assessed in another 10 rats from each group and in 10 normal rats.The survival time of donor hearts in the two EEPCW groups was significantly prolonged, to (15.9 +/- 2.4) days and (30.0 +/- 0.0) days, respectively, compared with (6.7 +/- 0.8) days in the control group. Pathological lesions in the two EEPCW groups were also less severe, and the percentages of CD3+, CD4+, and CD8+ lymphocytes and CD4+/CD8+ ratio were significantly lower in the EEPCW groups.Acute rejection of heart transplants and cellular immune reaction can be effectively suppressed using the EEPCW. Taking advantage of novel immunosuppressants derived from Chinese medicinal herbs used to treat abnormal pregnancy provides a hopeful road for future research and treatment in organ transplantation.